Coordinated Assembly of Ku and p460 Subunits of the DNA-dependent Protein Kinase on DNA Ends is Necessary for XRCC4–ligase IV Recruitment

Calsou, Patrick; Delteil, Christine; Frit, Philippe; Drouet, J; Salles, Bernard

doi:10.1016/s0022-2836(02)01328-1

Cited by 108 publications

(89 citation statements)

References 65 publications

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“…Therefore, we conclude that DNA-PKcs functions independently of Artemis in CSR. In this regard, DNA-PK might have an Artemis-independent NHEJ function in CSR and other processes, for example, by acting as scaffold for the XRCC4͞ ligase IV complex (20,(48)(49)(50)(51)(52). In addition, DNA-PK also may have functions outside of NHEJ in CSR (15,16).…”

Section: Discussionmentioning

confidence: 99%

Artemis-independent functions of DNA-dependent protein kinase in Ig heavy chain class switch recombination and development

Rooney

Alt

Sekiguchi

et al. 2005

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

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Assembly of Ig genes in B lineage cells involves two distinct DNA rearrangements. In early B cell development, site-specific double strand breaks (DSBs) at germ-line V, D, and J gene segments are joined via nonhomologous end-joining (NHEJ) to form variable region exons. Activated mature B cells can change expressed Ig heavy chain constant region exons by class switch recombination (CSR), which also involves DSB intermediates. Absence of any known NHEJ factor severely impairs joining of cleaved V, D, and J segments. In NHEJ, DNA-dependent protein kinase (DNA-PK), which is comprised of the Ku70͞80 end-binding heterodimer and the catalytic subunit (DNA-PKcs), activates Artemis to generate a nuclease that processes DSBs before ligation. Because inactivation of DNA-PKcs components also severely affects CSR, we tested whether DNA-PK also functions in CSR via activation of Artemis. To obviate the requirement for V(D)J recombination, we generated DNA-PKcs-and Artemis-deficient B cells that harbored preassembled Ig heavy chain and -light chain ''knock-in'' (HL) alleles. We found that Artemis-deficient HL B cells undergo robust CSR, indicating that DNA-PKcs functions in CSR via an Artemis-independent mechanism. To further elucidate potential Artemis-independent functions of DNA-PKcs, we asked whether the embryonic lethality associated with double-deficiency for DNA-PKcs and the related ataxia-telangiectasia-mutated (ATM) kinase was also observed in mice doubly deficient for ATM and Artemis. We found that ATM͞Artemis double-deficient mice were viable and born in normal Mendelian numbers. Therefore, we conclude that DNA-PKcs has Artemis-independent functions in CSR and normal development.ATM ͉ DNA-PKcs P recursor B and T cells assemble antigen receptor genes from component, germ-line V, D, and J segments via V(D)J recombination (1). V(D)J recombination is initiated by the lymphoid-specific RAG1͞2 endonuclease (RAG), which introduces DNA double strand breaks (DSBs) between V(D)J coding sequences and f lanking recombination signal (RS) sequences. RAG cleavage generates two distinct intermediates: hairpin coding ends and blunt RS ends, which are then fused via the nonhomologous end-joining (NHEJ) pathway of DSB repair (2). There are six known mammalian NHEJ factors (2).

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Section: Discussionmentioning

confidence: 99%

Artemis-independent functions of DNA-dependent protein kinase in Ig heavy chain class switch recombination and development

Rooney

Alt

Sekiguchi

et al. 2005

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

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“…All cells were grown in a humidified atmosphere, at 37°C with 5% CO 2 . Nuclear protein extracts were prepared as previously described (18), except that the final dialysis was performed for 3 h at 4°C in an excess volume of dialysis buffer as follows: 50 mM Tris-HCl, pH 7.5, 10% glycerol, 100 mM potassium glutamate, 1 mM EDTA, 1 mM dithiothreitol. After preparation, all of the extracts were immediately frozen and stored at Ϫ80°C.…”

Section: Methodsmentioning

confidence: 99%

“…One of the kinase functions is to regulate DNA end access to processing enzymes by means of an autophosphorylation operation (13)(14)(15). The XRCC4⅐DNA ligase IV complex is responsible for the ligation step (16,17), and both Ku and DNA-PKcs components are necessary to load this complex to the site of the break (18,19). Recently, a new core NHEJ factor with structural similarity to XRCC4, Cernunnos-XLF, has been identified concomitantly as deficient in a human RS-scid syndrome (20) and as an XRCC4-interacting protein (21).…”

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confidence: 99%

Interplay between Ku, Artemis, and the DNA-dependent Protein Kinase Catalytic Subunit at DNA Ends

Drouet

Frit

Delteil

et al. 2006

Journal of Biological Chemistry

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“…Although the role of the kinase activity remains to be fully elucidated, current evidence suggests that it regulates the process and facilitates processing of DNA ends (see below) [8]. The assembled DNA/DNA-PK complex recruits the second complex, which includes DNA ligase IV [9]. Until recently, this complex was thought to encompass two tightly associated proteins, DNA ligase IV and Xrcc4.…”

Section: Nhejmentioning

confidence: 99%

Contribution of DNA repair and cell cycle checkpoint arrest to the maintenance of genomic stability

2006

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Article (Unspecified) http://sro.sussex.ac.uk Jeggo, P. A. and Lobrich, M. (2006) Contribution of DNA repair and cell cycle checkpoint arrest to the maintenance of genomic stability. DNA Repair, 5 (9-10). pp. 1192 -1198 . ISSN 1568 This version is available from Sussex Research Online: http://sro.sussex.ac.uk/587/ This document is made available in accordance with publisher policies and may differ from the published version or from the version of record. If you wish to cite this item you are advised to consult the publisher's version. Please see the URL above for details on accessing the published version. Copyright and reuse:Sussex Research Online is a digital repository of the research output of the University.Copyright and all moral rights to the version of the paper presented here belong to the individual author(s) and/or other copyright owners. To the extent reasonable and practicable, the material made available in SRO has been checked for eligibility before being made available.Copies of full text items generally can be reproduced, displayed or performed and given to third parties in any format or medium for personal research or study, educational, or not-for-profit purposes without prior permission or charge, provided that the authors, title and full bibliographic details are credited, a hyperlink and/or URL is given for the original metadata page and the content is not changed in any way. DNA damage response mechanisms encompass pathways of DNA repair, cell cycle checkpoint arrest and apoptosis. Together, these mechanisms function to maintain genomic stability in the face of exogenous and endogenous DNA damage. ATM is activated in response to double strand breaks and initiates cell cycle checkpoint arrest. Recent studies in human fibroblasts have shown that ATM also regulates a mechanism of end-processing that is required for a component of double strand break repair. Human fibroblasts rarely undergo apoptosis after ionising radiation and, therefore, apoptosis is not considered in our review. The dual function of ATM raises the question as to how the two processes, DNA repair and checkpoint arrest, interplay to maintain genomic stability. In this review, we consider the impact of ATM's repair and checkpoint functions to the maintenance of genomic stability following irradiation in G2. We discuss evidence that ATM's repair function plays little role in the maintenance of genomic stability following exposure to ionising radiation. ATM's checkpoint function has a bigger impact on genomic stability but strikingly the two damage response pathways co-operate in a more than additive manner. In contrast, ATM's repair function is important for survival post irradiation.Introduction.

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Coordinated Assembly of Ku and p460 Subunits of the DNA-dependent Protein Kinase on DNA Ends is Necessary for XRCC4–ligase IV Recruitment

Cited by 108 publications

References 65 publications

Artemis-independent functions of DNA-dependent protein kinase in Ig heavy chain class switch recombination and development

Artemis-independent functions of DNA-dependent protein kinase in Ig heavy chain class switch recombination and development

Interplay between Ku, Artemis, and the DNA-dependent Protein Kinase Catalytic Subunit at DNA Ends

Contribution of DNA repair and cell cycle checkpoint arrest to the maintenance of genomic stability

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