Human Rad51C deficiency destabilizes XRCC3, impairs recombination, and radiosensitizes S/G2-phase Cells

Yi Ching Lio, David Schild, Mark A. Brenneman, J. Leslie Redpath, David J. Chen

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Abstract

The highly conserved Rad51 protein plays an essential role in repairing DNA damage through homologous recombination. In vertebrates, five Rad51 paralogs (Rad51B, Rad51C, Rad51D, XRCC2, and XRCC3) are expressed in mitotically growing cells and are thought to play mediating roles in homologous recombination, although their precise functions remain unclear. Among the five paralogs, Rad51C was found to be a central component present in two complexes, Rad51C-XRCC3 and Rad51B-Rad51C-Rad51D-XRCC2. We have shown previously that the human Rad51C protein exhibits three biochemical activities, including DNA binding, ATPase, and DNA duplex separation. Here we report the use of RNA interference to deplete expression of Rad51C protein in human HT1080 and HeLa cells. In HT1080 cells, depletion of Rad51C by small interfering RNA caused a significant reduction of frequency in homologous recombination. The level of XRCC3 protein was also sharply reduced in Rad51C-depleted HeLa cells, suggesting that XRCC3 is dependent for its stability upon heterodimerization with Rad51C. In addition, Rad51C-depleted HeLa cells showed hypersensitivity to the DNA-cross-linking agent mitomycin C and moderately increased sensitivity to ionizing radiation. Importantly, the radiosensitivity of Rad51C-deficient HeLa cells was evident in S and G2/M phases of the cell cycle but not in G1 phase. Together, these results provide direct cellular evidence for the function of human Rad51C in homologous recombinational repair.

Original languageEnglish (US)
Pages (from-to)42313-42320
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number40
DOIs
StatePublished - Oct 1 2004

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G2 Phase
S Phase
HeLa Cells
Genetic Recombination
Homologous Recombination
DNA
Recombinational DNA Repair
Proteins
Radiation Tolerance
Ionizing radiation
G1 Phase
Mitomycin
RNA Interference
Ionizing Radiation
Cell Division
Small Interfering RNA
DNA Damage
Adenosine Triphosphatases
Vertebrates
Cell Cycle

ASJC Scopus subject areas

  • Biochemistry

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Human Rad51C deficiency destabilizes XRCC3, impairs recombination, and radiosensitizes S/G2-phase Cells. / Lio, Yi Ching; Schild, David; Brenneman, Mark A.; Redpath, J. Leslie; Chen, David J.

In: Journal of Biological Chemistry, Vol. 279, No. 40, 01.10.2004, p. 42313-42320.

Research output: Contribution to journalArticle

Lio, Yi Ching ; Schild, David ; Brenneman, Mark A. ; Redpath, J. Leslie ; Chen, David J. / Human Rad51C deficiency destabilizes XRCC3, impairs recombination, and radiosensitizes S/G2-phase Cells. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 40. pp. 42313-42320.
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