Functional redundancy between repair factor XLF and damage response mediator 53BP1 in V(D)J recombination and DNA repair.
The classical nonhomologous DNA end-joining (C-NHEJ) double-strand break (DSB) repair pathway in mammalian cells maintains genome stability and is required for V(D)J recombination and lymphocyte development. Mutations in the XLF C-NHEJ factor or ataxia telangiectasia-mutated (ATM) DSB response protein cause radiosensitivity and immunodeficiency in humans. Although potential roles for XLF in C-NHEJ are unknown, ATM activates a general DSB response by phosphorylating substrates, including histone H2AX and 53BP1, which are assembled into chromatin complexes around DSBs. In mice, C-NHEJ, V(D)J recombination, and lymphocyte development are, at most, modestly impaired in the absence of XLF or ATM, but are severely impaired in the absence of both. Redundant functions of XLF and ATM depend on ATM kinase activity; correspondingly, combined XLF and H2AX deficiency severely impairs V(D)J recombination, even though H2AX deficiency alone has little impact on this process. These and other findings suggest that XLF may provide functions that overlap more broadly with assembled DSB response factors on chromatin. As one test of this notion, we generated mice and cells with a combined deficiency for XLF and 53BP1. In this context, 53BP1 deficiency, although leading to genome instability, has only modest effects on V(D)J recombination or lymphocyte development. Strikingly, we find that combined XLF/53BP1 deficiency in mice severely impairs C-NHEJ, V(D)J recombination, and lymphocyte development while also leading to general genomic instability and growth defects. We conclude that XLF is functionally redundant with multiple members of the ATM-dependent DNA damage response in facilitating C-NHEJ and discuss implications of our findings for potential functions of these factors.
Department of Genetics, Harvard Medical School, Children's Hospital, Immune Disease Institute, Howard Hughes Medical Institute, Boston, MA 02115, USA.
Valentyn
V
Oksenych
Frederick W
FW
Alt
Vipul
V
Kumar
Bjoern
B
Schwer
Duane R
DR
Wesemann
Erica
E
Hansen
Harin
H
Patel
Arthur
A
Su
Chunguang
C
Guo
D000818:Animals; D002868:Chromosomal Proteins, Non-Histone / Q000502:physiology*; D004249:DNA Damage; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000502:physiology*; D051379:Mice; D000071857:Tumor Suppressor p53-Binding Protein 1; D060152:V(D)J Recombination
D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; C498302:Trp53bp1 protein, mouse; D000071857:Tumor Suppressor p53-Binding Protein 1; C519508:XLF protein, mouse