DNA repair deficiency and neurological disease

REVIEWS

DNA repair deficiency and neurological disease Peter J. McKinnon

Abstract | The ability to respond to genotoxic stress is a prerequisite for the successful development of the nervous system. Mutations in various DNA repair factors can lead to human diseases that are characterized by pronounced neuropathology. In many of these syndromes the neurological component is among the most deleterious aspects of the disease. The nervous system poses a particular challenge in terms of clinical intervention, as the neuropathology associated with these diseases often arises during nervous system development and can be fully penetrant by childhood. Understanding how DNA repair deficiency affects the nervous system will provide a rational basis for therapies targeted at ameliorating the neurological problems in these syndromes.

Department of Genetics and Tumour Cell Biology, St Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, Tennessee 38105, USA. e‑mail: [email protected] doi:10.1038/nrn2559 Published online 15 January 2009; corrected online 27 January 2009.

Maintaining genomic integrity through DNA repair is fundamental to the functioning and survival of an organism. Compromising genomic DNA can lead to a range of human disorders that exhibit developmental defects, immune deficiency and cancer. In particular, the nervous system is often profoundly affected by DNA repair deficiency, which can result in neurodegen­ eration, microcephaly or brain tumours. More broadly, defective DNA repair in mature neural tissues has also been linked to aging and, recently, to common neuro­ degenerative syndromes such as Alzheimer’s disease and Parkinson’s disease1–4. The developmental dynamics of the nervous system require vigilant DNA repair processes that can repair multiple types of damage. DNA repair is especially cru­ cial during the early period of rapid proliferation, in which progenitors expand and differentiate to gener­ ate the nervous system. In the mature nervous system, DNA repair is also essential to prevent DNA damage arising from sources such as oxidative metabolism from blocking active transcription. Understanding the interplay between the signalling networks that maintain genomic stability in the nervous system will be of para­ mount importance for treating diseases associated with DNA repair deficiency. This Review discusses the role of DNA repair during the genesis of the nervous system, and how this process maintains neural homeostasis.

DNA integrity during neural development Many human DNA repair deficiency syndromes are con­ genital and it is clear that the developing nervous sys­ tem can be markedly affected by DNA repair deficiency.

Understanding neural development is therefore impor­ tant for determining how DNA repair deficiencies affect the nervous system. Detailed reviews that summarize neural development are available (see REFS 5–9). In humans, the nervous system begins to form during the first trimester of gestation and continues to form until after birth. The complexity of the mature nervous system results from a relatively simple strategy of proliferation, differentiation and migration (FIG. 1). The engine that drives neural development is rapid cellular proliferation in the ventricular and subventricular zones that line the four ventricles. These regions contain proliferating neural stem and progenitor cells5,6,9, and DNA replica­ tion in these cells is of paramount importance as they will populate the nervous system and remain in place for the life of the organism. If these stem and progenitor cells incur mutation, the expansion of these damaged cells could subsequently lead to disease. Thus, during this phase of proliferative expansion DNA repair is of the utmost importance. Neural progenitors also proliferate in the subventricular zone, the lateral ventricle and the dentate gyrus throughout adulthood9. Although it is unclear exactly how this process contributes to the form and function of the mature human nervous system, evidence from mice indicates that ongoing adult neuro­ genesis is functionally important, suggesting that DNA repair is important in these areas throughout life10,11. The bulk of nervous system cells are either neurons or glia, but there are a myriad of specialized cell types in these broad classes, with specific functional roles that support the divergent functions of neural tissue. These cellular subgroups are characterized by their

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REVIEWS Ventricular zone Stem and progenitor cells

Subventricular zone Differentiation and migration

Mature nervous system Specialized neurons and glia

Proliferating Differentiating Types of damage

Strand breaks Transcriptional interference

DNA damage-induced apoptosis Replicative stress

Oxidative metabolism

Types of DSB repair (HR and NHEJ) Helicases etc. repair

DSB repair (NHEJ) SSB repair NER

SSB repair DSB repair NER

Figure 1 | DNA damage and repair during nervous system development. During development the nervous system forms through widespread Nature proliferation, and Reviewsmigration | Neuroscience differentiation. The diversity of the nervous system comes from stem cells and progenitor cells that divide in the ventricular zone and, albeit to a lesser extent, in the subventricular zone and then undergo differentiation, migration and maturation to give rise to the neurons and glia of the adult nervous system. A wide variety of functionally specialized cells with unique properties originate from proliferative cells in one of four ventricles in the nervous system. At different stages of development the nervous system is susceptible to different types of DNA damage. During proliferation, replication-associated DNA strand breaks can occur that may require DNA double strand break (DSB) repair, involving homologous recombination (HR) or non-homologous end joining (NHEJ) as well as the associated functions of helicases and other replication components that interface with DNA repair. In differentiating cells, repair options are more limited as HR is not available. At this stage, NHEJ repairs DNA DSBs whereas other types of DNA damage require nucleotide excision repair (NER) or single strand break (SSB) repair. Because the nervous system can easily replace cells during development, DNA damage-induced apoptosis is also a frequent outcome at these stages. In the mature nervous system, strand breaks and DNA modification from oxidative damage engage SSB repair, NER and DSB repair, and DNA damage that is not repaired can disrupt transcription and lead to cell death.

morphology, the neurotransmitters that they use for cellular communication and the connections that they form with other cells. This diversity extends throughout the nervous system, and increases the requirement for specific DNA repair pathways. Oxidative load The amount of oxidative stress, in the form of free radicals or reactive oxygen species, that is encountered by a tissue.

Free radical A molecule that can be produced by metabolism and that contains unstable and reactive unpaired electrons that can damage cellular components such as DNA.

Helical distortion A topological perturbation of the DNA double helix that the DNA repair machinery can register as damage.

Endogenous sources of DNA damage because of its complexity, the nervous system presents a substantial challenge for delineating the spatiotemporal requirements for DNA repair. one of the main questions associated with inherited human DNA repair deficiency syndromes is the nature and source of endogenous DNA damage in the nervous system. The rapid cellular proliferation that occurs during development is associated with replication­induced dam­ age, and therefore an intact DNA repair system is needed at this stage. However, DNA repair remains important after replication ends and maturation commences12–14. The high oxidative load of the brain, and the free radicals produced by cellular metabolism, can lead to many dif­ ferent types of DNA damage, and so it is possible that a

limited number of aetiological agents are responsible for generating diverse lesions. There is an increase in DNA breaks after high oxidative load, such as that induced by overexpression of superoxide dismutase or increased oxygen tension15, and depending on the developmen­ tal timing, these strand breaks can lead to apoptosis14. Additionally, oxidative stress­induced strand breaks in DNA can also affect transcription16,17. In addition to DNA strand breaks, oxidative stress can generate specific types of DNA lesions that can only be repaired by the excision of the damaged section (see below)18. DNA damage can also be a by­product of normal neural functioning: activation of ionotropic glutamate receptors leads to the formation of γH2AX (a phosphorylated form of the histone variant H2AX), a marker of DNA damage and repair 19. Regardless of the exact nature of the endogenous lesions, the vast reper­ toire of DNA damage response factors and the many human DNA repair deficiency syndromes underscore the need to respond to a wide variety of insults (FIG. 2). Although the focus of this Review is the link between DNA repair deficiency and neurological disease, nor­ mal DNA repair processes can also be involved in neuropathology. In the triplet­repeat­expansion class of neurodegenerative diseases, such as those that involve polyglutamine tracts in encoded proteins, oxidative stress in the brain has been shown to cause DNA glycosylase­ mediated expansion of triplet repeats towards diseases levels20,21. Thus, as well as oxidative stress being causal in DNA repair deficiency syndromes, it could be respon­ sible for activating repair systems that inadvertently expand triplet sequences, also leading to disease.

DNA damage responses DNA can undergo various modifications, including strand breaks, base damage, helical distortions and strand cross links. biochemically distinct DNA repair path­ ways have evolved to repair each of these specific lesions (FIG. 2). basic strategies for DNA repair involve excising the damaged region — by removing small numbers of nucleotides through the base excision repair (beR) pathway or by removing longer stretches through the nucleotide excision repair (NeR) pathway. In the case of DNA strand breaks, either single strand break (SSb) or double strand break (DSb) repair pathways address the damage. Defects in these pathways can lead to human neurological disease, with the resultant neuropathology often linked to the type of damage and the develop­ mental stage at which it occurs (TABLE 1). Several recent reviews discuss the detailed biochemistry of DNA repair pathways linked to human diseases22–28. Here, we will consider the main DNA damage pathways and examine the impact of defects in these pathways and how they lead to neuropathology. DSB repair pathways and related diseases one of the earliest­recognized syndromes linking DNA damage and neurodegeneration was ataxia telangiecta­ sia (AT) (discussed below). Children with AT have severe neurodegeneration and an extreme sensitivity to ionizing radiation29–31. AT established a compelling

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REVIEWS Replication stress Oxidative damage Chemotherapeutics Radiation

Base modifications

Cross link

SSB

Single-stand repair Base-excision repair

DSB Bulky lesion

Nucleotide-excision repair

Non-homologous end joining Homologous recombination

Examples of associated human neurological diseases AOA1 AOA2 SCAN1

XP CS TTD

A-T ATLD NBS

Figure 2 | Types of DNA damage and repair. Various types of DNA damage can occur in neural cells as a result of endogenous agents, such as replication stress or| Neuroscience free radicals Nature Reviews from oxidative metabolism. Exogenous agents, such as ionizing or ultraviolet radiation and chemotherapeutics, can also cause different types of DNA damage. These agents can cause single strand breaks (SSBs) or double strand breaks (DSBs) in the DNA, base modifications, helix-distorting bulky lesions or cross links of DNA strands. Biochemically distinct DNA repair pathways are available to repair each class of DNA damage. DNA repair pathways that are particularly important for nervous system function are those that repair SSBs and DSBs and nucleotide excision repair. When any of these pathways is defective, diseases can result that affect the nervous system; representative examples of human syndromes linked to defects in the particular DNA repair pathways are listed. Defective repair of SSBs can lead to ataxia with oculomotor apraxia 1 (AOA1), AOA2 or spinocerebellar ataxia with axonal neuropathy 1 (SCAN1), whereas defects in nucleotide excision repair can lead to xeroderma pigmentosum (XP), Cockayne Syndrome (CS) or trichothiodystrophy (TTD). Defective responses to DSBs can lead to ataxia telangiectasia (AT), AT-like disease (ATLD) or Nijmegen breakage syndrome (NBS).

Non-homologous end joining (NHEJ). One of two distinct biochemical pathways for the repair of DNA DSBs. NHEJ modifies non-compatible termini and directly ligates the broken DNA ends.

Homologous recombination (HR). One of two distinct biochemical pathways for the repair of DNA DSBs. HR functions in S or G2 of the cell cycle and uses sister chromatid DNA as an undamaged template.

Hypomorphic mutation A mutation that does not fully eliminate the function of a gene product and that typically results in a less severe phenotype than a loss-of-function mutation.

link between faulty responses to DNA DSbs and neuro­ degeneration. Although this connection was recognized in the mid 1970s32, it was not until 1995 that AT was found to result from the mutation of a single gene — ataxia telangiectasia, mutated (ATM), which encodes a kinase33. Subsequently, the DNA DSb signalling pathway controlled by this protein kinase was elucidated34,35. DNA DSB repair pathways. DNA DSbs initiate a sig­ nalling cascade that leads to the repair and resolution of the break or to apoptosis. The repair of DNA DSbs occurs through either non-homologous end joining (NHeJ) or homologous recombination (HR) (FIG. 3). These two major repair pathways maintain the integrity of DNA after DSbs caused by endogenous or exogenous agents, and inactivation of either NHeJ or HR results in marked defects in nervous system development14. Although both pathways repair DSbs, they are biochemically distinct: HR operates in proliferating cells whereas NHeJ can function in both proliferating and differentiating cells. HR is an error­free process that uses a sister chroma­ tid as a template to achieve precise repair 36,37. Homology­ directed repair initially involves DSb processing by

the MRe11–RAD50–Nijmegen breakage syndrome 1 (NbS1; also known as NbN) (MRN) complex in col­ laboration with bRCA1 C­terminal interacting protein (CTIP; also known as RbbP8). This generates 3′ sin­ gle­stranded DNA that becomes bound by replication protein A (RPA) and the recombinase RAD51, which facilitates homology search and strand invasion, a pro­ cess whereby template DNA from a sister chromatid is inserted into the damaged chromatid and acts as an error­free template38–40, and effects recombinational repair in collaboration with accessory factors 36,37,41 (FIG. 3). During development, this pathway maintains the genomic integrity of neural progenitor cells. In non­replicating cells, such as those in the mature nervous system, DNA DSbs are repaired by NHeJ42 (FIG. 3). In contrast to HR, NHeJ modifies the two DNA ends so that they are compatible for direct ligation42–44. ligation of the DNA ends is catalysed by DNA ligase IV (lIG4) in conjunction with its binding partner XRCC4 after end modification by the DNA protein kinase com­ plex (DNA­PKCS (also known as PRKDC), KU70 (also known as XRCC6) and KU80 (also known as XRCC5)) and associated end­processing factors44. An additional factor, Cernunnos (also known as NHeJ1 and Xlf), has been identified as a binding partner of the lIG4– XRCC4 complex and is also necessary for efficient liga­ tion through NHeJ45,46. Hypomorphic mutations in lIG4 that lead to attenuated ligase activity have been associ­ ated with a disorder termed lIG4 syndrome47, which is characterized by radiosensitivity, immunodeficiency and microcephaly 48, and defective Cernunnos also leads to microcephaly46 (TABLE 1). The severity of lIG4 syndrome is related to the amount of residual lIG4 activity 49. DSB signalling. Coincident with DNA DSb repair is the activation of specific signalling events that initiate cell cycle arrest to stop proliferation and allow DNA repair to proceed. ATM is a key protein kinase that functions at the apex of a signalling pathway that coordinates cell cycle arrest after DNA damage34. ATM­dependent phos­ phorylation of cell cycle checkpoint effectors, such as p53, checkpoint kinase 2 (CHeK2), structural maintenance of chromosomes 1A (SMC1A), breast cancer associated 1 (bRCA1) and NbS1, activates G1, intra­S and G2–M checkpoints (FIG. 4). DNA DSbs also induce rapid phos­ phorylation of histone H2AX, facilitating the retention of numerous proteins that assemble at the break, including NbS1, mediator of DNA damage checkpoint 1 (MDC1) and p53 binding protein 1 (P53bP1)50–52. Detailed bio­ chemical descriptions of ATM signalling after DNA damage are available35,52–57. In the developing nervous system, apoptosis is a common outcome after DNA DSbs and might in fact be preferable to DNA repair, as cells can easily be replaced from the abundant progenitor population that is present during neural development 14. ATM is an important determinant of DSb­induced apoptosis in neural pro­ genitor cells exiting the cell cycle58, as it signals to the apoptosis machinery through CHeK2 and p53 to acti­ vate pro­apoptotic gene expression and apoptosis59–61. Thus, the absence of ATM can result in the viability of

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REVIEWS Table 1 | Human DNA repair deficiency syndromes Disease or syndrome

Gene

Neurological symptoms

extraneurological symptoms

DNA DSB repair deficiency Ataxia telangiectasia

ATM

Ataxia, neurodegeneration, telangiectasia and dysarthria

Immunological defects, malignancy and sterility

Ataxia telangiectasia-like disorder

MRE11

Ataxia, neurodegeneration, dysarthria and oculomotor apraxia

Mild immunological defects

Nijmegen breakage syndrome

NBS1

Microcephaly

Immunological defects and lymphoid malignancy

ATR-Seckel syndrome

ATR

Microcephaly and mental retardation

Growth defects

LIG4 syndrome

LIG4

Microcephaly

Developmental/growth delay, immunodeficiency and lymphoma

Human immunodeficiency with microcephaly

Cernunnos

Microcephaly

Immunodeficiency

Fanconi anaemia

BRCA2

Microcephaly and medulloblastoma

Bone marrow and congenital defects

TDP1

Ataxia, neurodegeneration, peripheral axonal motor Hypercholesterolaemia and and sensory neuropathy, and muscle weakness hypoalbuminaemia

Ataxia with oculomotor apraxia 1 APTX

Ataxia, neurodegeneration, oculomotor apraxia and Hypercholesterolaemia and peripheral neuropathy hypoalbuminaemia

DNA SSB repair deficiency Spinocerebellar ataxia with axonal neuropathy

NER deficiency Xeroderma pigmentosum

XPA–XPG

Neurodegeneration and microcephaly

UV sensitivity and skin cancer

Cockayne syndrome

CSA, CSB, XPB, XPD and XPG

Microcephaly and dysmyelination

Progeria and otherwise variable presentation

Trichothiodystrophy

XPD, XPB and TTD-A

Neurodevelopmental defects and dysmyelination

Brittle hair and otherwise variable presentation

FAA–FAL

Microcephaly and medulloblastoma (brain tumours in FANCD2 and FANCN subtypes)

Anaemia, developmental defects and cancer

Werner syndrome

WRN

?

Severe progeria and cancer

Rothmund Thomson syndrome

RTS

?

Cancer

Bloom syndrome

BLM

?

Proportional dwarfism and cancer

Ataxia, neurodegeneration and oculomotor apraxia

Absent or minimal

DNA cross link repair Fanconi anaemia

Helicase deficiency

Ataxia with oculomotor apraxia 2 SETX

APTX, aprataxin; ATM, ataxia telangiectasia, mutated; ATR, ataxia telangiectasia and RAD3-related; BRCA2, breast cancer associated 2; DSB, double strand break; LIG4, ligase IV, NHEJ1, non-homologous end joining 1; SSB, single strand break; TDP1, tyrosyl-DNA phosphodiesterase 1; UV, ultraviolet; ?, neurological deficits are not clearly defined.

Cell cycle checkpoint A biochemical signalling event, activated following stimuli such as DNA damage, that pauses the cell cycle to allow time for recovery from the insult and to maintain cellular homeostasis.

DNA­damaged cells that would normally have been elim­ inated by apoptosis. This feature of ATM loss might be linked to the neuropathology of AT, as a failure to eliminate these damaged cells could later lead to cell loss. An ATM­related protein kinase, ataxia telangiecta­ sia and RAD3­related (ATR) also participates in DNA damage signalling 62. ATM and ATR are both crucial for nervous system function and phosphorylate many common substrates, including p53, NbS1, bRCA1 and CHeK1, in response to DNA damage34,35,52,55, and recent estimates suggest that these kinases can phosphorylate 700 or more proteins in response to ionizing radiation63. However, ATM and ATR also have non­redundant physiological roles. In the mouse, inactivation of ATR is lethal during early development 64,65, whereas mice that lack ATM are viable and, although tumour­prone, can often have a normal lifespan. Hypomorphic mutations of ATR can lead to ATR­Seckel syndrome, a disease that

presents with developmental defects and microcephaly, rather than the progressive neurodegeneration that is characteristic of AT25,30,48. Nonetheless, both kinases can cooperatively respond to DSbs during S phase62,66. In addition to ATM and ATR, another related kinase — the catalytic subunit of DNA­PK (DNA­PKCS), a central factor in NHeJ — is also important for signal­ ling after DNA damage67. Although NHeJ is clearly important in the nervous system, roles for DNA­PKCS outside of NHeJ are not clear. However, it may have some functional redundancy with ATM, as only dual inactivation of ATM and DNA­PKCS is lethal early during embryogenesis68. Disorders related to defective DSB repair and signalling. The hallmark feature of AT is neurodegeneration, and individuals with AT are typically wheelchair­bound by their early teen years. other features of AT include

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REVIEWS Double strand breaks

a Homologous recombination

Single strand breaks

b Non-homologous end joining

DNA DSB

c SSB repair or BER

DNA DSB

RPA

Damaged base

DNA SSB

DNA glycosylase

RAD51 KU70 and KU80 PARP1

APTX

Sister chromatid APE1 PARP1

BRCA2 XRCC2

XLF XRCC4

Polβ

TDP1

XRCC1

LIG4

LIG3

DNA-PK

FEN1

XR CC 1

LIG

3

Polδ/ε

PC

Short patch repair

NA

LIG1

LIG

1

Sister chromatid

Long patch repair

Figure 3 | repairing DNA strand breaks. The repair of DNA double strand breaks (DSBs) and single strand breaks (SSBs) Nature Reviews | Neuroscience occurs by distinct biochemical pathways. Two different repair pathways can deal with a DSB, and which does depends on the proliferative state of the cell. a | Homologous recombination can be used in proliferating cells. Replication protein A (RPA) coats the resected single stranded DNA and recruits RAD51 recombinase which, together with a multitude of other factors36,37,41, including breast cancer associated 2 (BRCA2) and XRCC2 (REFS 36,37,41), repairs the DNA through a processes in which template DNA from the sister chromatid is inserted into the damaged chromatid and acts as an error-free template. Ligation of the break requires DNA ligase I (LIG1). b | Non-homologous end joining (NHEJ) can function in proliferating and non-proliferating cells. Heterodimeric KU70 (also known as XRCC6) and KU80 (also known as XRCC5) bind DNA ends and recruit DNA-PK (the catalytic subunit of the DNA-dependent kinase) which, together with XRCC4, Cernunnos (also known as NHEJ1) and DNA ligase IV (LIG4), reseals the DNA ends after suitable end processing from various nucleases42–44. Because some nucleotides may be lost in this process, NHEJ is often referred to as error-prone repair. However, DNA breaks are repaired, and so these errors might generally be of little consequence in non-dividing cells such as neurons. c | SSBs can arise from damaged bases being removed by specific DNA glycosylases (referred to as base excision repair (BER)) or from direct backbone damage that severs the DNA strand. In both cases the stand break leads to poly ADP-ribose polymerase 1 (PARP1) accumulation, which facilitates recruitment of the XRCC1 scaffolding protein that is important for promoting repair. XRCC1 recruits repair factors that modify the DNA ends for ligation. Depending on the nature of the DNA ends present at the SSB, tyrosyl DNA phosphodiesterase 1 (TDP1) (which acts on 3′ modified ends or topoisomerase I DNA adducts) or aprataxin (APTX) (which acts on 5′ ends resulting from abortive ligation events) may be required to modify the damaged DNA for repair. Repair can involve the removal of a single nucleotide (short-patch repair) or a longer patch of nucleotides (long-patch repair) by PCNA and LIG1. APE1, apurinic/apyrimidinic endonuclease 1; FEN1, flap structure-specific endonuclease 1; LIG3, DNA ligase III; pol, DNA polymerase.

immune dysfunction, sterility, extreme radiosensitivity and cancer predisposition29–31. Patients with AT mani­ fest muscle hypotonia, truncal swaying while sitting or standing and abnormalities of eye movement, features that are typical of cerebellar dysfunction69. Atrophy of the cerebellar folia, widespread loss of Purkinje cells, granule cell loss and significant thinning of the molecular layer

are characteristic defects found in the AT cerebellum70–72. MRI and computed tomography (CT) studies also show cerebellar atrophy that is especially prevalent in ver­ mal regions73–75. Patients with AT also exhibit marked alterations in deep tendon reflexes, loss of the ability to sense vibration, reduction in sensory conduction veloc­ ity and axonal degeneration of peripheral nerves69,76,77.

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REVIEWS widespread neurodegeneration also occurs in other regions of the nervous system, but this is variable and usually late­stage69. Two syndromes that phenotypically overlap with AT are associated with mutations of components of the MRN complex (FIG. 4). These are ataxia telangiectasia­like dis­ ease (ATlD) and Nijmegen breakage syndrome (NbS), which result from mutations in MRe11 and NbS1, respectively 78–80. In contrast to AT, the disease­causing mutations in ATlD and NbS are hypomorphic (as the MRN complex is indispensable for DNA replication and cellular survival), and although they disable some impor­ tant MRN functions, the protein nonetheless retains the essential activity that supports DNA replication. Some individuals originally diagnosed with AT were later shown to carry hypomorphic mutations in MRe11 (REFS 79,80) and therefore to have ATlD. This find­ ing demonstrated the interrelationship between ATM and the MRN complex, and the role of DNA damage in the aetiology of AT. The clinical features of ATlD are similar to those of AT, although patients with ATlD do not have telangiectasia and have a later­onset ataxia phenotype with a slower progression of neurodegenera­ tion80. NbS was described as an AT variant because of its close similarity to AT81,82. However, NbS is character­ ized by microcephaly rather than neurodegeneration83,84, with occasional medulloblastoma brain tumours85,86. Mutations of the NBS1 gene in NbS lead to the synthe­ sis of a truncated protein missing the amino terminus87 that can interact with MRe11 and RAD50 and can carry out most of the functions of NbS1 that are required for normal embryonic development. The interrelationship of AT, ATlD and NbS results from the requirement of the MRN complex for normal ATM activation after DNA DSbs88,89 (FIG. 4). Although the exact details of ATM activation by the MRN complex are not fully understood, the function of MRN as a sensor of DNA damage provides insight. MRN binds to a DNA break and undergoes a conformational change that leads to RAD50 binding and forming a tether between MRN complexes bound to opposing DNA strands, facilitat­ ing DNA repair 90–92. The ability of the MRN complex to tether broken DNA ends provides a physical basis for the recruitment of crucial signalling kinases, such as ATM. The association of ATM with the MRN complex involves it interacting with the NbS1 C­terminal region93–95, and thus NbS1 is required for the localization of activated ATM at sites of DSbs96.

SSB repair pathways and related disease A mechanistic counterpoint to the repair of DSbs is the use of distinct biochemical factors that deal with damage to only one DNA strand (FIG. 3). DNA SSbs are common DNA lesions and can occur owing to the direct effects of reactive oxygen species or indirectly as an interme­ diate during beR22,97; each of these lesions is repaired by similar core machinery 98. The repair of a damaged base involves the action of a lesion­specific DNA glyco­ sylase and an endonuclease that initiates base excision, followed by apurinic site incision by the APeX1 endo­ nuclease, recruitment of repair factors through XRCC1

DNA damage S NBS1 SMC1 FANCD2 ATM G1

p53 p21

MRN

CHEK1 BRCA1 CDC25C

G2

p53 and CHEK2 M Apoptosis

Figure 4 | ATM signalling in response to DNA damage. Ataxia telangiectasia, mutatedNature (ATM) is a serine/threonine Reviews | Neuroscience protein kinase modulated by the MRE11–RAD50– Nijmegen breakage syndrome 1 (NBS1; also known as NBN) (MRN) complex that is critically important for responding to DNA double strand breaks (DSBs), and loss of these factors can lead to profound neuropathology. Many ATM substrates are important cell cycle and apoptotic regulators. A primary function of ATM after DNA damage is checkpoint activation. At each phase of the cell cycle ATM activates checkpoint proteins, among which are Fanconi anaemia group D2 (FANCD2), structural maintenance of chromosomes 1 (SMC1), NBS1, cell division cycle 25C (CDC25C), checkpoint kinase1 (CHEK1) and breast cancer associated 1 (BRCA1); for a full list of ATM substrates, see REFS 34,35. Key ATM substrates are p53 and CHEK2, which are responsible for activating the G1 checkpoint proteins and apoptosis. A main function of ATM in the nervous system may be to modulate DNA damage-induced apoptosis. This is because the prolific cell division that occurs in the developing nervous system allows damaged cells to be easily replaced, and so ATM has an important genome-monitoring function as immature cells exit the cell cycle, whereby cells with DNA damage are eliminated in an ATM-dependent manner14.

and poly ADP­ribose polymerase (PARP), end modifica­ tion and gap filling by DNA polymerase, and ligation by ligase III (short­patch repair) or ligase I in collaboration with PCNA (long­patch repair)22,98–100. Defects in DNA SSb repair are associated with ataxia with oculomotor apraxia 1 (AoA1) and spinocerebellar ataxia with axonal neuropathy (SCAN1), neurodegen­ erative syndromes that are similar to AT26,101–105 (TABLE 1). The neurological presentation of AoA1 is also almost identical to that of AT, leading to AoA1 initially being classified as an AT variant106. However, SSb repair defects lead almost exclusively to neuropathology without the extraneurological phenotypes that are associated with DSb repair deficiency. This is probably due to the avail­ ability of backup repair pathways in proliferating and other tissues107. The phenotypes associated with AoA1 and SCAN1 are similar, but oculomotor apraxia is absent from SCAN1 (REFS 104,108).

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REVIEWS Complementation group A subgroup of a disease in which the causative defective multiprotein complex can be functionally corrected by reintroducing a missing or defective component from cellular extracts of another individual with the same disease.

The genes that are mutated in AoA1 and SCAN1 are aprataxin (APTX) and tyrosyl­DNA phosphodiesterase 1 (TDP1), respectively. TDP1 repairs altered 3′ DNA ends arising from oxidative damage and topoisomerase 1– DNA covalent complexes22,107,109–113. Neural cells from a Tdp1–/– mouse have a pronounced defect in repairing camptothecin­induced topoisomerase 1–DNA cleavage and hydrogen peroxide­induced SSbs114,115. APTX is a member of the histidine triad superfamily of nucleo­ tide hydrolases and possesses an AMP–lysine hydro­ lase activity, which is required for repairing 5′­AMP intermediates that arise from failed DNA ligation reactions26,116,117. Although APTX could in principal be involved in DSb repair 118, the lack of extraneurologi­ cal features in AoA1 strongly suggests that it is caused by a primary defect in SSb repair 26,101,103, as its pheno­ type contrasts with syndromes associated with defec­ tive DSb responses, such as AT or lIG4 syndrome, which are characterized by multi­systemic defects. furthermore, the fact that oxidative stress, commonly present in the brain, results in many more SSbs than DSbs also suggests that SSbs are a likely aetiological agent causing AoA1. Although it is rare, AoA1 is the most common reces­ sive ataxia in Japan and the second most common of all autosomal recessive ataxias in Portugal103. These SSb repair deficiency syndromes highlight the unique sus­ ceptibility of the nervous system to DNA damage, and despite the lack of extraneurological features, the neuro­ pathology in these diseases is severe106,119. However, the interrelationship between DNA repair deficiency and neuropathology in these diseases is intricate and the full details remain to be elucidated (BOX 1).

Repairing other types of DNA damage The NER pathway. In contrast to the DNA strand­break repair mechanisms discussed above, the NeR pathway Box 1 | Neuropathology and DNA repair defects Studies of the neuropathology of DNA repair deficiency syndromes aim to understand the cause and consequence of DNA damage and thereby provide a rational basis for therapies to ameliorate the neurological deficits. In general, loss of DNA repair capacity affects development, and neurological issues are often apparent early in childhood. One brain region that is particularly susceptible to many DNA repair deficiency syndromes is the cerebellum. Notably, the cerebellum continues its development for approximately a year after birth. During this period cerebellar neurogenesis is substantial, and the granule neurons generated become the most numerous neuronal type in the brain7. Diseases such as ataxia telangiectasia (AT), ataxia with oculomotor apraxia 1 (AOA1), AOA2 and spinocerebellar ataxia with axonal neuropathy 1 (SCAN1) all profoundly affect the cerebellum. The underlying mutations in each disease affect the repair of DNA single or double strand breaks (see main text for details). Although the neuropathologies of these diseases have parallels, the phenotypes indicate differences in the timing or types of DNA damage that presumably reflect the specific activities of the affected enzyme. Analysis of AT reveals cerebellar atrophy with widespread loss of purkinje and granule cells30. In AOA1, cerebellar atrophy is present but the loss of purkinje cells is restricted to the cerebellar flocculus116. Features of AT and AOA1 are oculomotor apraxia and early onset, whereas in SCAN1 and AOA2 disease onset is later, and in SCAN1 oculomotor apraxia is absent. We will need to know how specific types of DNA damage spatiotemporally affect different neural cell types before we can understand the molecular basis of the phenotypic range of DNA repair deficiency syndromes.

resolves DNA­distorting lesions, typified by those that are generated by ultraviolet (UV) radiation. This repair pathway is functionally linked to transcription, as the basal TfIIH transcription and repair complex, which is required for RNA polymerase II­dependent tran­ scription, is partly comprised of proteins that are also required for NeR23,120–122. NeR repair of DNA during transcription is termed transcription­coupled repair (TCR), whereas repair that occurs independently of tran­ scription is termed global genomic repair (GGR) (FIG. 5). These pathways use common repair factors and differ only in the mode of DNA damage recognition. During GGR, it is the XPC or XPe (also known as DDb1) com­ plexes that initially recognize DNA damage28,122–124. If the lesion occurs in a transcribed gene, it is sensed as a blockage to RNA polymerase II and requires CSb (also known as eRCC6) (and CSA (also known as eRCC8)) to initiate repair 23. for both pathways, DNA damage verification and DNA unwinding is carried out by TfIIH (complexed with the XPb (also known as eRCC3) and XPD (also known as eRCC2) helicases) together with XPG (also known as eRCC5), in a pre­incision com­ plex that also involves XPA125–129. Incisions are then made either side of the damage site by the nuclease action of XPf and XPG130,131, and excision of the lesion is followed by gap filling by DNA polymerase δ or ε and strand rejoining by ligase I or III. Detailed reviews outlining the biochemistry of this pathway are available23,28,122,132,133. The distinction between these two branches of NeR is important because neuropathology in NeR syndromes is associated with disabled TCR132,134. Xeroderma pigmentosum and related disorders. Mutations in various components of the NeR pathway can lead to the human syndromes xeroderma pigmen­ tosum (XP), Cockayne syndrome (CS), and trichothio­ dystrophy (TTD) 132,133,135. Although these diseases are clinically distinct, they do show some phenotypic similarities. In fact, in some cases different mutations in the same NeR protein can lead to different syndromes (FIG. 5). eight different XP complementation groups (arising from mutations in different XPA–XPG NeR compo­ nents or from expression of a variant polymerase, XPV) are known. Around 30% of individuals with XP develop neurodegeneration that results in global brain atrophy 135,136. Neurological symptoms appear from around 4 years of age and involve cognitive impairment followed by cerebellum­related problems, such as dys­ arthria, balance disturbances and sensorineural hearing loss; progressive neuropathy is evident from the second decade135,137,138. Increased cognitive impairment and corticospinal degeneration occur progressively with age, and at death almost all XPA individuals have severe neurological problems137,139. This neuropathology is observed in individuals from XP subgroups in which the mutations affect TCR132,134. Notably, XPC individu­ als who have defective GGR but not TCR have little overt neurological impairment, although mild brain atrophy is present 137 and a brain tumour has been reported140.

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REVIEWS Transcription-coupled repair

Global genomic repair

CSB

Damage recognition

+CSA CSB

XPE

XPC

Pre-incision complex forms

CSA Damage verification and helix unwinding TFIIH XPB XPD Pol II TTDA XPG RPA XPA

Xeroderma pigmentosum XPA–XPG

Damage excision TFIIH XPB XPD XPG XRF RPA XPA

Cockayne syndrome CSA,CSB, XPB, XPD and XPG Trichothiodystrophy XPB, XPD and TTDA

Repair synthesis Polδ/ε

LIG1/3

Figure 5 | Nucleotide excision repair and related diseases. Defects in nucleotide excision repair (NER) lead to at least three human syndromesNature characterized Reviews | by Neuroscience neurodegeneration: xeroderma pigmentosum, Cockayne syndrome and trichothiodystrophy (TABLE 1). NER can involve transcription-coupled repair (TCR), which occurs when the transcription complex encounters a damaged template, and the more general repair pathway global genomic repair (GGR), which repairs lesions present in non-transcribed DNA. The difference between these pathways is at the step of damage detection. The sequence of events in NER involves DNA damage recognition followed by an incision, damage removal and repair synthesis. DNA damage recognition in TCR involves initiation by CSB (also known as ERCC6) after the RNA polymerase II complex encounters damage. In GGR damage is recognized by XPE (also known as DDB1) or XPC. Pre-incision events involve XPA or replication protein A (RPA). The TFIIH complex, including the XPB (also known as ERCC3) and XPD (also known as ERCC2) helicases, unwinds the DNA helix, resulting in an open conformation. XPG (also known as ERCC5) is then recruited to the complex and binds to TFIIH and RPA. The Cockayne syndrome proteins, CSA and CSB, are also involved in TCR and might shift TFIIH from a transcription to a repair complex. Dual incision involves XPG performing the initial cleavage 3′ to the site of DNA damage, followed by 5′ cleavage through the action of the XPF nuclease. Repair synthesis is performed by DNA polymerase (pol) δ or ε with PCNA, and ligation to seal the gap involves DNA ligase I (LIG1). Three main classes of diseases resulting from disruption of NER are listed, and the pathway components that are disrupted in the respective diseases are represented in corresponding colours. TTDA, trichothiodystrophy group A protein.

DNA repair defects in CS involve the TCR compo­ nent of NeR, and individuals with CS show progres­ sive and severe neuropathology. It should be noted that the neuropathology in CS is distinct from that which is seen in XP. The neurological problems that are present in CS include profound microcephaly, mental retardation and progressive brain atrophy. Importantly, dysmyelination and calcification of the basal ganglia are also characteristic of CS but not XP141,142. Two CS complementation groups result from mutations in two different genes, CSA and CSB, which promote the

repair of DNA lesions encountered by the polymer­ ase II complex. both CSA, which encodes a wD­ repeat­containing protein143, and CSb, which encodes a helicase144, interact with TfIIH121,143. CS can also result from certain mutations in XPb, XPD or XPG28,135,145. because the neuropathology differs between XP and CS, it is likely that something additional to defective TCR underlies one of the diseases. The mutations that lead to CS probably result in more general and substantial transcriptional defects than those that are involved in XP. Another possibility is that in CS there is a defect in a sub­branch of GGR termed domain­ associated repair, which is important for repair of the non­transcribed strand of genes and has been suggested to be particularly important in neurons133,146. ongoing studies will elaborate the interplay between NeR factors and the basis for the resultant phenotypes when specific components are disrupted. TTD is also associated with defective TCR. It arises from certain mutations of the XPD helicase (or, more rarely, XPb or trichothiodystrophy group A (TTDA) protein) and is characterized by sulphur­deficient brit­ tle hair and phenotypic similarities to XP and CS135. Sensitivity to UV light is observed in ~50% of TTD individuals but, similar to CS, cancer predisposition is absent. The neurological abnormalities observed in TTD include microcephaly, mental retardation, deafness and ataxia147. like with CS, a striking aspect of the neuro­ pathology of TTD is dysmyelination135,141,148. Recently, the neurological defects associated with TTD were shown to involve deregulated expression of brain­specific thyroid hormone receptor­dependent genes134. These results suggested that defects in the thyroid hormone pathway are a major cause of TTD, which is consistent with defi­ cits associated with hypomyelination, learning disorders and motor dysfunction. The pleiotropic nature of TTD­ associated neurological defects probably results from impairment of other transcription factors in addition to thyroid hormone signalling. further insights into the impact of NeR on brain function have come from mouse models in which NeR genes have been disrupted149. Inactivation of XPG caused a loss of cerebellar Purkinje cells150, and dual inactiva­ tion of CSb and XPA led to cerebellar atrophy and loss of granule cells151, underscoring the importance of this pathway for brain function.

Other disorders related to DNA repair deficits Various other syndromes, including fanconi anaemia (fA), werner syndrome (wS), bloom syndrome (blM) and Rothmund Thomson syndrome (RTS), also result from DNA repair defects (TABLE 1). fA can result from disruption to any one of a large number of DNA repair factors required to remove cross links in DNA strands, and currently there are 12 fA complementation groups152–154. fA presents with similar neuropathology to NbS, with microcephaly as a hallmark feature. Some subgroups of fA (fANC D1 and fANC N) result from mutations in bRCA2 or its interacting partner PAlb1 (also known as TTR) that render DNA DSb repair defec­ tive152,155,156. furthermore, brain tumours are a feature of

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REVIEWS the D1 subgroup157. Given the link between DNA repair deficiency and tumourigenesis, it is somewhat surpris­ ing that brain tumours do not occur more frequently in DNA repair deficiency syndromes (BOX 2). blM, wS and RTS result from defective ReCQl4­ related helicase function 158–161. The resultant neuro­ pathology in these syndromes is not well characterized, although the syndromes produce quite dramatic and varying pathologies outside of the nervous system, including cancer, progeria and proportional body size defects. In the case of wS, a neurological involve­ ment has been reported but is controversial162–164. why mutations in these helicases do not more substantially affect the nervous system is not clear. The role of the helicases in replication and their link to HR suggest that they have an important function in genomic­integrity checks during neural precursor proliferation. Perhaps neurological disease is avoided in these syndromes by apoptotic elimination of damaged neural progenitors during development, or perhaps the helicases are not central to neural stem and progenitor replication and maintenance. However, mutations in another helicase — the ortho­ logue of the yeast Sen1p helicase, termed senataxin — result in AoA2, a syndrome characterized by pro­ nounced cerebellar degeneration, oculomotor apraxia and sensory motor neuropathy 165–170. Additionally, dif­ ferent mutations in this gene can lead to a form of amyo­ trophic lateral sclerosis171. AoA2 is characterized by a specific defect in the response to certain types of DNA DSbs172, but is not sensitive to ionizing radiation173. Although distinct DNA repair pathways respond to specific DNA lesions, interplay between the different Box 2 | Neurodegeneration versus brain tumours Given the link between DNA damage and tumourigenesis, it might be expected that DNA repair deficiency syndromes that affect the nervous system would also present with brain tumours. Although this does happen85,86,157, it is more frequent in these syndromes that tumours occur in tissues outside of the brain. This probably reflects the tissue-specific consequences of gene loss. For example, xeroderma pigmentosum (Xp) results from defects in different genes in the nucleotide excision repair (NER) pathway, which can lead to cancer predisposition, neurodegeneration or both. Defects that lead to neuropathology generally affect transcription-coupled repair132,134, whereas in tumour-prone individuals with Xp, NER defects affect global genomic repair (GGR). During nervous system development, DNA damage that would normally be repaired by GGR, which in individuals with Xp could lead to tumours, might instead be eliminated by apoptosis. However, brain tumours can occur at low frequency in Xp140. Syndromes resulting from DNA strand-break response defects, such as ataxia telangiectasia (AT), reflect the tissue-specific signalling role for ataxia telangiectasia, mutated (ATM), in which a primary function is to activate apoptosis of non-replicating DNA-damaged neural cells58. In the immune system, which is where tumours mostly arise in AT, ATM maintains the fidelity of immunoglobin DNA rearrangements, preventing oncogenic rearrangements53. However, other DNA repair deficiency syndromes, such as Fanconi anaemia (the FANC D1 subgroup with mutations in breast cancer associated 2), develop medulloblastoma brain tumours (located in the cerebellum)157, reflecting the occurrence of DNA damage in rapidly proliferating cells of the developing cerebellum. This is also the case for Nijmegen breakage syndrome (NBS), in which loss of NBS1 leads to microcephaly and can also result in medulloblastoma85,86. In these cases, medulloblastoma might occur because the DNA repair defect is quite substantial and affects the rapidly proliferating and abundant granule neuron precursors, which have a relatively high chance of acquiring mutations leading to transformation.

DNA repair pathways is likely to fine­tune the DNA damage response. for example, pathways that deal with DNA interstrand cross links to prevent fA can also be involved in the repair of DSbs154, and SSb repair factors also interact with DSb repair factors174, suggesting that there are important interactions and crosstalk between DNA repair pathways. Unravelling the biology of DNA repair pathways and their role in disease prevention will benefit from ongoing work using mouse genetics. The biochemical defect for many human diseases characterized by ataxia or microcephaly is unknown175, although in some cases sensitivity to DNA damaging agents suggests that the cause is a defect in DNA repair 176. Thus, it is likely that the range of DNA repair deficiency diseases will continue to expand, as the respective disease­causing mutations are uncovered.

Tissue specificity of repair pathways Historically, information about DNA repair pathways that affect the nervous system has been obtained from individuals with DNA repair deficiency syndromes. Using cells derived from patients with AT, XP or fA, defects in the response to ionizing radiation, UV light and cross­linking agents were identified, and these cells have continued to provide important biochemi­ cal insights into these DNA repair pathways. However, with an increased understanding of the molecular basis of the DNA damage response, and with the develop­ ment of mouse models of DNA repair deficiency, it has become apparent that many of the key signalling path­ ways that underpin the response to DNA damage exhibit a striking tissue or cell­type specificity 177. for example, differential radiosensitivity was found in the brains of mice lacking ATM: immature postmitotic neural cells were resistant to infrared radiation whereas proliferat­ ing progenitors were susceptible to it 58,60. Inactivation of NbS1 in the brain markedly affected the cerebel­ lum178, whereas loss of MDC1, despite its central role in the DNA damage response179,180, resulted in a rela­ tively modest phenotype181. These findings illustrate the challenges of translating in vitro biochemical data to a physiological setting. Neural sensitivity to DNA damage The nervous system is very sensitive to DNA damage, particularly in comparison with other tissues that con­ tain non­replicating cell types. for example, in DNA SSb repair deficiencies the neurological symptoms are almost the exclusive presentation of the disease. There are some unique properties of the nervous system that may account for this relative sensitivity. The brain metabolizes ~20% of consumed oxygen but has a lower capacity than other body parts to neutralize reactive oxy­ gen species, and neurons are particularly susceptible to oxidative stress182. This high oxidative load presumably generates free radicals, which will increase DNA damage, particularly DNA SSbs, in the mature nervous system. Therefore, cells with defective SSb repair will be more susceptible to this insult. As SSbs could subsequently interfere with the transcriptional machinery, this could eventually result in cell death16,132,183.

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REVIEWS The cerebellum is a prominent target of many human DNA repair deficiency syndromes, raising the question of why this organ is so uniquely sensitive to DNA damage (BOX 1). A notable feature of the cerebellum is its extended postnatal development 7. This period of rapid cell prolif­ eration will generate replication stress­associated DNA damage that might affect granule neurons and, perhaps indirectly, other cerebellar cell types. Purkinje cells in the cerebellum are also sensitive to oxidative stress184,185, and antioxidants can promote the survival of cultured ATM­deficient Purkinje cells186. There are many unresolved issues regarding the apparent vulnerability of the nervous system. because DNA repair deficiency syndromes are congenital it is important to know what periods during neural develop­ ment are vulnerable and what the ongoing requirements for DNA repair pathways in mature neural compartments are. Some DNA repair deficiency syndromes have a later onset, and it is unknown whether this reflects a require­ ment for DNA repair in postmitotic neurons or whether it is a result of accrued developmental damage.

Mitochondrial DNA repair The mitochondria have a key role in nervous system function187,188. Mitochondrial DNA (mtDNA) encodes 13 proteins that are part of the respiratory chain and 24 RNAs, although proteins encoded by nuclear genes are also present in this organelle. Damage to the mtDNA can readily occur, as the mitochondrial genome is located on the inner membrane, a major site of reac­ tive oxygen species generation189. The primary DNA repair mechanism that is operative in the mitochondria is the beR and SSb repair pathway, and there are many specific mitochondrial versions of components of this pathway (which are encoded by nuclear DNA) 190,191. Various mtDNA lesions, including point mutations and large­scale deletions of mtDNA, can lead to mito­ chondrial dysfunction and cellular apoptosis191,192. In the nervous system, compromised mitochondrial func­ tion has been linked to neurodegeneration, including Alzheimer’s and Parkinson’s disease1,187,189,191. Therefore, DNA repair defects that affect mtDNA could conceiv­ ably be a significant harbinger of neurological disease. It will therefore be important to determine the con­ tribution of mitochondrial dysfunction to the neuro­ pathology of human syndromes resulting from DNA repair defects. Insights from neuropathology Generally speaking, the consequence of DNA repair defects is microcephaly or neurodegeneration. Although both pathologies ultimately result from cell loss, the fun­ damental causes are different. Microcephaly results from cell loss or proliferation defects during neurogenesis. These could reflect increased apoptosis resulting from a failure to repair replication errors or DNA damage incurred during differentiation. Data from mouse mod­ els indicate that defects in DSb repair, such as those that result from lIG4 or bRCA2 deficiency, markedly affect neurogenesis by causing widespread apoptosis during development that can result in microcephaly12,13,193. This is

probably the case in human microcephaly — for example, that which is a feature of lIG4 syndrome48. DNA repair is important for the maintenance of stem cells, and loss of these could also contribute to microcephaly 194,195. Stem cells might be a particularly susceptible population in other syndromes that cause microcephaly, such as ATR­ Seckel syndrome, as defects in ATR are known to affect replication, leading to cell loss or cell cycle arrest62,194–196. This could also be the case for fA­associated microceph­ aly, as the fA pathway is required for the maintenance of neural stem and progenitor cells during development and in the adult197. The neurodegenerative phenotype that is typified by AT and other diseases with similar clinical presentation, such as AoA1 and ATlD, contrasts with the microceph­ aly that is seen in ATR­Seckel syndrome and fA. early neurological problems in AT point to a requirement for ATM during nervous system development. This may relate to the need for ATM to remove cells with DNA damage early after cell cycle exit to prevent their incor­ poration into the nervous system58,60. Diseases such as AoA2 and SCAN1 generally have a later onset, pos­ sibly because backup DNA repair protects cells during development whereas postmitotic neurons succumb to unrepaired damage, probably as a result of transcriptional interference16,22. Additionally, some subgroups of XP (notably XPA) also show severe neurodegeneration, but in many cases are also associated with microcephaly 137. This neuropathology might reflect developmental neuro­ genesis defects coupled with ongoing cell loss in the mature brain. Thus, cell loss arising from DNA damage during development leads to microcephaly, whereas the progressive degenerative syndromes might reflect cell loss after birth as a consequence of accumulated damage during development, or progressive loss from ongoing DNA damage that affects transcription.

Conclusions and perspectives Many different cell types are required for the func­ tion of the nervous system, and this diversity presents a challenge for understanding how the dynamics of the DNA damage response ensure homeostasis in the brain. DNA repair deficiency syndromes are congenital, and thus they raise important developmental questions, such as how much of the eventual neuropathology is already set in place at birth. The consequences of DNA dam­ age and associated neuropathology may involve other aspects of cellular homeostasis. for example, mitochon­ drial dysfunction is increasingly being linked to neuro­ logical disease, and as mitochondrial DNA is susceptible to damage, DNA repair defects could substantially affect this organelle in the brain192,198. Additionally, the mecha­ nism of cell death in neurodegenerative syndromes is not fully understood and could involve abortive cell cycle re­ entry, as has been suggested for cell loss in Alzheimer’s disease and as has been reported for AT199–201. Therefore, these additional aspects of DNA damage may be impor­ tant contributors to neuropathology in DNA repair deficiency syndromes. future challenges include finding answers to the following questions: are neurons and glia equally

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REVIEWS susceptible to the loss of DNA damage responses? what is the basis for the differential impact of defective repair in specific cell types and different neural tissues? what is the ongoing requirement for DNA repair and what are the relevant DNA repair pathways in mature neu­ ral compartments? If oxidative stress is important as an

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Acknowledgements

I thank the US National Institutes of Health and the American Lebanese Syrian Associated Charities of St Jude Children’s Research Hospital for financial support, and laboratory members for discussions and comments. I also thank an anonymous reviewer for providing helpful suggestions regarding NER diseases. Space constraints limited the number of primary research papers cited.

DATABASES Entrez Gene: http://www.ncbi.nlm.nih.gov/entrez/query. fcgi?db=gene APEX1 | APTX | ATM | ATR | Cernunnos | CSA | CSB | CTIP | DNA-PKCS | KU70 | KU80 | LIG4 | ligase I | ligase III | MDC1 | MRE11 | NBS1 | P53BP1 | PALB1 | PARP | PCNA | RAD50 | RAD51 | RECQL4 | RPA | senataxin | TDP1 | XPA | XPB | XPC | XPD | XPE | XPG | XRCC1 | XRCC4 OMIM: http://www.ncbi.nlm.nih.gov/entrez/query. fcgi?db=OMIM Alzheimer’s disease | AOA1 | AOA2 | AT | ATLD | LIG4 syndrome | NBS | Parkinson’s disease | SCAN1

FURTHER INFORMATION Information on AT from the Neuromuscular Disease Center at Washington University: http://neuromuscular.wustl.edu/ataxia/dnarep.html Database of allelic variations in XP genes: http://xpmutations.org Database of mutations associated with Werner syndrome: http://www.pathology.washington.edu/research/werner/ database/index.html AT children’s project: http://www.atcp.org National organization for rare disorders: http://www.rarediseases.org/ Bloom’s syndrome foundation: http://www.milogladsteinfoundation.org/#foundersjump Fanconi Anaemia Research Fund: http://www.fanconi.org/ Xeroderma Pigmentosum (XP) Society: http://www.xps.org/ Cockayne Syndrome Network: http://cockaynesyndrome.net/main/ All liNks Are AcTive iN The oNliNe pDf

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