Different types of V(D)J recombination and end-joining defects in DNA double-strand break repair mutant mammalian cells

Verkaik, Nicole S.; Lange, Rebecca Esveldt-van; Heemst, Diana van; Brüggenwirth, Hennie T.; Hoeijmakers, Jan H.J.; Zdzienicka, Małgorzata Z.; Gent, Dik C. van

doi:10.1002/1521-4141(200203)32:3<701::aid-immu701>3.0.co;2-t

Cited by 171 publications

(126 citation statements)

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“…Canonical NHEJ involves the successive intervention of the KU80-KU70 heterodimer, DNA-PKcs-Artemis, and, finally, ligase IV (Lig4) associated with its cofactors XRCC4 and Cernunnos/Xlf (2,3). KU-independent NHEJ (KU-alt) has been described in vitro in both acellular extracts and cultured cells (1,(4)(5)(6).…”

mentioning

confidence: 78%

“…Alternative, XRCC4-independent DSB repair pathways (XRCC4-alt) have also been described, using episomic plasmid in cultured cells, using pulse field gel electrophoresis, or in in vitro biochemical experiments (5)(6)(7)(8)(9)(10). One hypothesis could propose that XRCC4 and KU are implicated in the same canonical NHEJ pathway, whereas the alternate pathway is independent of both KU and XRCC4.…”

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confidence: 80%

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Defects in XRCC4 and KU80 differentially affect the joining of distal nonhomologous ends

Guirouilh‐Barbat

Rass

Plo

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

152

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XRCC4-null mice have a more severe phenotype than KU80-null mice. Here, we address whether this difference in phenotype is connected to nonhomologous end-joining (NHEJ). We used intrachromosomal substrates to monitor NHEJ of two distal doublestrand breaks (DSBs) targeted by I-SceI, in living cells. In xrcc4-defective XR-1 cells, a residual but significant end-joining process exists, which primarily uses microhomologies distal from the DSB. However, NHEJ efficiency was strongly reduced in xrcc4-defective XR-1 cells versus complemented cells, contrasting with KU-deficient xrs6 cells, which showed levels of end-joining similar to those of complemented cells. Nevertheless, sequence analysis of the repair junctions indicated that the accuracy of end-joining was strongly affected in both xrcc4-deficient and KU-deficient cells. More specifically, these data showed that the KU80/XRCC4 pathway is conservative and not intrinsically error-prone but can accommodate non-fully complementary ends at the cost of limited mutagenesis.double-strand break repair ͉ genome rearrangements D NA double-strand breaks (DSBs) are harmful lesions generated by a variety of endogenous or exogenous stresses, potentially leading to genomic rearrangement. Nonhomologous endjoining (NHEJ) is a prominent pathway for DSB repair (1). Canonical NHEJ involves the successive intervention of the KU80-KU70 heterodimer, DNA-PKcs-Artemis, and, finally, ligase IV (Lig4) associated with its cofactors XRCC4 and Cernunnos/Xlf (2, 3). KU-independent NHEJ (KU-alt) has been described in vitro in both acellular extracts and cultured cells (1, 4-6).Alternative, XRCC4-independent DSB repair pathways (XRCC4-alt) have also been described, using episomic plasmid in cultured cells, using pulse field gel electrophoresis, or in in vitro biochemical experiments (5-10). One hypothesis could propose that XRCC4 and KU are implicated in the same canonical NHEJ pathway, whereas the alternate pathway is independent of both KU and XRCC4. However, in transgenic mice, the inactivation of XRCC4 or Lig4 results in a more severe phenotype than the inactivation of KU (11), suggesting that XRCC4 might have an additional essential function; however, studies show that XRCC4 and Lig4 do not have roles outside of NHEJ, whereas in contrast, KU acts in other processes such as transcription, apoptosis, and responses to the cell microenvironment (12)(13)(14).Alternatively, these varying phenotypes in mice may actually result from differences in DSB repair efficiencies, indicating that defects in XRCC4 might be more deleterious for DSB repair than defects in KU. What challenges this hypothesis, however, is that substantial class switch recombination (CSR) has recently been shown to occur in mouse B cells without XRCC4, whereas no CSR was recorded in cells devoid of KU (15-18).The relative contributions of XRCC4 and KU80 versus the XRCC4-alt and KU-alt pathways, respectively, to DSB repair remain unclear in wild-type cells. Contrasting results were obtained in living cells, using an episomic plasm...

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confidence: 78%

mentioning

confidence: 80%

Defects in XRCC4 and KU80 differentially affect the joining of distal nonhomologous ends

Guirouilh‐Barbat

Rass

Plo

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

152

183

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“…This backup system relies mainly on patches of a few base pairs of (micro)homology, that may assist in alignment of the ends (Kabotyanski et al, 1998;Verkaik et al, 2002). Although microhomology-mediated end-joining is not very efficient for V(D)J recombination and repair of blunt DSBs (van Heemst et al, 2004), this backup pathway can mediate Ig heavy chain class switch recombination in B cells rather efficiently (Yan et al, 2007).…”

Section: Alternative End-joining Pathwaysmentioning

confidence: 99%

“…The precise analysis of V(D)J recombination products is also very informative. V(D)J recombination substrates can be transfected into patient cells (primary fibroblasts) or residual junctions from bone marrow precursor B-cells can be amplified (Table 2) (Verkaik et al, 2002;van der Burg et al, 2006van der Burg et al, , 2007. Artemis deficiency causes normal signal joint formation, but a strong reduction in the numbers of coding joints, which are characterized by long stretches of P-nucleotides due to aberrant hairpin opening (Rooney et al, 2003;van der Burg et al, 2007).…”

Section: Intersection Of Nhej With Other Cellular Processesmentioning

confidence: 99%

Non-homologous end-joining, a sticky affair

2007

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Rejoining of broken chromosomes is crucial for cell survival and prevention of malignant transformation. Most mammalian cells rely primarily on the non-homologous end-joining pathway of DNA double-strand break (DSB) repair to accomplish this task. This review focuses both on the core non-homologous end-joining machinery, which consists of DNA-dependent protein kinase and the ligase IV/XRCC4 complex, and on accessory factors that facilitate rejoining of a subset of the DSBs. We discuss how the ATM protein kinase and the Mre11/Rad50/Nbs1 complex might function in DSB repair and what role ionizing radiation-induced foci may play in this process.

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“…Additionally, WRN exonuclease activity is required to fully compliment a Werner syndrome DNA-end joining phenotype, in an in vivo plasmid based assay . The in vivo data did not define a specific cellular pathway, but the elevated microhomology-mediated repair observed in WRN exonuclease deficient cells is similar to the phenotypes associated with essential NHEJ proteins (Melek et al, 1998,Verkaik et al, 2002, possibly linking WRN to this pathway. However, Werner syndrome cells have mild radiation sensitivity, which rules out WRN as an essential DSB repair protein, but WRN exonuclease may nevertheless be used for resolution of a limited class of DSBs.…”

Section: Double-strand Breaks Base Excision Repair and Wrnmentioning

confidence: 99%

Developing master keys to brain pathology, cancer and aging from the structural biology of proteins controlling reactive oxygen species and DNA repair

2007

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This review is focused on proteins with key roles in pathways controlling either reactive oxygen species or DNA damage responses, both of which are essential for preserving the nervous system. An imbalance of reactive oxygen species or inappropriate DNA damage response likely causes mutational or cytotoxic outcomes, which may lead to cancer and/or aging phenotypes. Moreover, individuals with hereditary disorders in proteins of these cellular pathways have significant neurological abnormalities. Mutations in a superoxide dismutase, which removes oxygen free radicals, may cause the neurodegenerative disease amyotrophic lateral sclerosis. Additionally, DNA repair disorders that affect the brain to varying extents include ataxia-telangiectasia-like disorder, Cockayne syndrome or Werner syndrome. Here, we highlight recent advances gained through structural biochemistry studies on enzymes linked to these disorders and other related enzymes acting within the same cellular pathways. We describe the current understanding of how these vital proteins coordinate chemical steps and integrate cellular signaling and response events. Significantly, these structural studies may provide a set of master keys to developing a unified understanding of the survival mechanisms utilized after insults by reactive oxygen species and genotoxic agents, and also provide a basis for developing an informed intervention in brain tumor and neurodegenerative disease progression.

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Different types of V(D)J recombination and end-joining defects in DNA double-strand break repair mutant mammalian cells

Cited by 171 publications

References 45 publications

Defects in XRCC4 and KU80 differentially affect the joining of distal nonhomologous ends

Defects in XRCC4 and KU80 differentially affect the joining of distal nonhomologous ends

Non-homologous end-joining, a sticky affair

Developing master keys to brain pathology, cancer and aging from the structural biology of proteins controlling reactive oxygen species and DNA repair

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