Ku Regulates the Non-Homologous End Joining Pathway Choice of DNA Double-Strand Break Repair in Human Somatic Cells

被引:184
作者
Fattah, Farjana [1 ]
Lee, Eu Han [1 ]
Weisensel, Natalie [1 ]
Wang, Yongbao [1 ]
Lichter, Natalie [1 ]
Hendrickson, Eric A. [1 ]
机构
[1] Univ Minnesota, Sch Med, Dept Biochem Mol Biol & Biophys, Minneapolis, MN 55455 USA
来源
PLOS GENETICS | 2010年 / 6卷 / 02期
关键词
DEPENDENT PROTEIN-KINASE; MISMATCH REPAIR; POLY(ADP-RIBOSE) POLYMERASE; V(D)J RECOMBINATION; ERROR-PRONE; X-RAY; HOMOLOGOUS RECOMBINATION; IONIZING-RADIATION; GENOMIC STABILITY; TELOMERE LENGTH;
D O I
10.1371/journal.pgen.1000855
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The repair of DNA double-strand breaks (DSBs) is critical for the maintenance of genomic integrity and viability for all organisms. Mammals have evolved at least two genetically discrete ways to mediate DNA DSB repair: homologous recombination (HR) and non-homologous end joining (NHEJ). In mammalian cells, most DSBs are preferentially repaired by NHEJ. Recent work has demonstrated that NHEJ consists of at least two sub-pathways-the main Ku heterodimer-dependent or "classic'' NHEJ (C-NHEJ) pathway and an "alternative'' NHEJ (A-NHEJ) pathway, which usually generates microhomology-mediated signatures at repair junctions. In our study, recombinant adeno-associated virus knockout vectors were utilized to construct a series of isogenic human somatic cell lines deficient in the core C-NHEJ factors (Ku, DNA-PKcs, XLF, and LIGIV), and the resulting cell lines were characterized for their ability to carry out DNA DSB repair. The absence of DNA-PKcs, XLF, or LIGIV resulted in cell lines that were profoundly impaired in DNA DSB repair activity. Unexpectedly, Ku86-null cells showed wild-type levels of DNA DSB repair activity that was dominated by microhomology joining events indicative of A-NHEJ. Importantly, A-NHEJ DNA DSB repair activity could also be efficiently de-repressed in LIGIV-null and DNA-PKcs-null cells by subsequently reducing the level of Ku70. These studies demonstrate that in human cells C-NHEJ is the major DNA DSB repair pathway and they show that Ku is the critical C-NHEJ factor that regulates DNA NHEJ DSB pathway choice.
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页数:14
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