Replication Protein A Physically Interacts with the Bloom's Syndrome Protein and Stimulates Its Helicase Activity

Robert M. Brosh, Ji-Liang Li, Mark K. Kenny, Julia K. Karow, Marcus P. Cooper, Raichal P. Kureekattil, Ian D. Hickson, Vilhelm A. Bohr
2000 Journal of Biological Chemistry  
Bloom's syndrome is a rare autosomal recessive disorder characterized by genomic instability and predisposition to cancer. BLM, the gene defective in Bloom's syndrome, encodes a 159-kDa protein possessing DNAstimulated ATPase and ATP-dependent DNA helicase activities. We have examined mechanistic aspects of the catalytic functions of purified recombinant BLM protein. Through analyzing the effects of different lengths of DNA cofactor on ATPase activity, we provide evidence to suggest that BLM
more » ... nslocates along singlestranded DNA in a processive manner. The helicase reaction catalyzed by BLM protein was examined as a function of duplex DNA length. We show that BLM catalyzes unwinding of short DNA duplexes (<71 base pairs (bp)) but is severely compromised on longer DNA duplexes (>259-bp). The presence of the human singlestranded DNA-binding protein (human replication protein A (hRPA)) stimulates the BLM unwinding reaction on the 259-bp partial duplex DNA substrate. Heterologous single-stranded DNA-binding proteins fail to stimulate similarly the helicase activity of BLM protein. This is the first demonstration of a functional interaction between BLM and another protein. Consistent with a functional interaction between hRPA and the BLM helicase, we demonstrate a direct physical interaction between the two proteins mediated by the 70-kDa subunit of RPA. The interactions between BLM and hRPA suggest that the two proteins function together in vivo to unwind DNA duplexes during replication, recombination, or repair.
doi:10.1074/jbc.m001557200 pmid:10825162 fatcat:hfbbhxsnsjh4vetg6jbzf2dbcu