Ionizing Radiation Triggers Chromatin-bound kin17 Complex Formation in Human Cells*

The human DNA-binding HSAkin17 protein cross-reacts with antibodies raised against the stress-activated Escherichia coli RecA protein. We show here that HSAkin17 protein is directly associated with chromosomal DNA as judged by cross-linking experiments on living cells. We detected increased amounts of DNA-bound HSAkin17 protein 24 h after γ irradiation, with 2.6-fold moreHSAkin17 molecules after 6 Gy of irradiation (46,000–117,000 molecules). At this time we observed that highly proliferating RKO cells displayed the concentration and co-localization of HSAkin17 and replication protein A in nucleoplasmic foci. Our results suggest that 24 h post-irradiationHSAkin17 protein may localize at the sites of unrepaired DNA damages. RKO clones expressing an HSA KIN17antisense transcript (RASK.5 and RASK.13 cells) revealed that reducedHSAkin17 protein levels are correlated with a decrease in clonogenic cell growth and cell proliferation, as well as an accumulation of cells in early and mid-S phase. Taken together our observations support the idea that HSAkin17 protein is a DNA maintenance protein involved in the cellular response to the presence of DNA damage and suggest that it helps to overcome the perturbation of DNA replication produced by unrepaired lesions.

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