Inhibition of homologous recombination by variants of the catalytic subunit of the DNA-dependent protein kinase (DNA-PKcs).
Two major DNA double-strand break repair pathways exist in all eukaryotes, nonhomologous DNA end joining (NHEJ) and homologous recombination (HR). Although both pathways can function throughout the cell cycle, NHEJ predominates in G0/G1) (when a replicated sister chromatid is unavailable), whereas HR makes a more substantial contribution in S and G2. How a cell chooses between these two important DNA repair pathways is largely unknown. DNA-dependent protein kinase (DNA-PK) is critical for NHEJ. Here, we describe two conserved splice variants of a catalytic subunit of DNA-PK (DNA-PKcs) that are expressed predominately in nondividing cells. Although both encode stable products, neither reverses the NHEJ defects in DNA-PKcs-deficient cells. In fact, cells expressing one of the DNA-PKcs variants are slightly more radiosensitive than cells completely deficient in DNA-PKcs. We investigated whether cells expressing the DNA-PKcs variants had any other DNA repair deficits and found that these cells are considerably more sensitive to both etoposide and mitomycin C than cells that express no DNA-PKcs at all. Because repair of DNA damage induced by these two agents requires intact HR, we tested whether the NHEJ-defective variants of DNA-PKcs inhibit double-strand break-induced HR in an integrated substrate. In cells expressing the NHEJ-defective variants, HR was markedly reduced. Because the splice variants are expressed highly only in nondividing cells, quiescent cells would be afforded a mechanism to inhibit repair by means of HR when sister chromatids are not available as templates for accurate repair with low risk of genome rearrangement, thereby enhancing genome stability.
Convery
Erin
E
Department of Pathobiology and Diagnostic Investigation, Michigan State University, East Lansing, MI 48824, USA.
Shin
Euy Kyun
EK
Ding
Qi
Q
Wang
Wei
W
Douglas
Pauline
P
Davis
Laurie S
LS
Nickoloff
Jac A
JA
Lees-Miller
Susan P
SP
Meek
Katheryn
K
D017398:Alternative Splicing; D020134:Catalytic Domain; D002453:Cell Cycle / Q000235:genetics; D002455:Cell Division; D002460:Cell Line; D045744:Cell Line, Tumor; D017124:Conserved Sequence; D051747:DNA-Activated Protein Kinase; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000528:radiation effects; D014644:Genetic Variation; D006801:Humans; D009687:Nuclear Proteins; D010957:Plasmids; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000528:radiation effects; D021122:Protein Subunits / Q000235:genetics; D011839:Radiation, Ionizing; D011995:Recombination, Genetic; D014162:Transfection
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D021122:Protein Subunits; D051747:DNA-Activated Protein Kinase; C497263:PRKDC protein, human; D017346:Protein Serine-Threonine Kinases