Identification and characterization of a novel nuclear structure containing members of the homologous recombination and DNA damage response pathways

Abstract

The human RAD9A protein is required for successful execution of the G2/M DNA damage checkpoint. Along with RAD1 and HUS1, RAD9A exists in a heterotrimeric ring-shaped complex which is necessary for activation of the CHK1 checkpoint kinase. RAD9A is also required for proper localization of both TopBP1 and the Claspin adaptor protein during the DNA damage response. We have shown large, RAD9A-dense nuclear foci containing several members of the homologous recombination pathway as well as BRCA1 and the DNA damage marker γH2AX. This RAD9A-dense body is closely associated with the inactive X in HeLa cells but not in other cell types analyzed including a Klinefelter's syndrome-derived line containing multiple Xi. We have also shown these foci disappear after cell synchronization but are enriched after treatment with the homologous recombination inhibitor pentoxifylline. We conclude these foci are the result of an active process, suspended in perturbed cells, that involves interaction between the cell cycle checkpoint and homologous recombination machinery.

Keywords: 911 complex; DNA repair; Homologous recombination; RAD9A.