Human RPA activates BLM’s bidirectional DNA unwinding from a nick
Zhenheng Qin,
Lulu Bi,
Xi-Miao Hou,
Siqi Zhang,
Xia Zhang,
Ying Lu,
Ming Li,
Mauro Modesti,
Xu-Guang Xi,
Bo Sun
Affiliations
Zhenheng Qin
School of Life Science and Technology, ShanghaiTech University, Shanghai, China; Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China; University of Chinese Academy of Sciences, Beijing, China
Lulu Bi
School of Life Science and Technology, ShanghaiTech University, Shanghai, China
Xi-Miao Hou
College of Life Sciences, Northwest A&F University, Yangling, China
Siqi Zhang
School of Life Science and Technology, ShanghaiTech University, Shanghai, China; Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China; University of Chinese Academy of Sciences, Beijing, China
Xia Zhang
School of Life Science and Technology, ShanghaiTech University, Shanghai, China
Ying Lu
Institute of Physics, Chinese Academy of Sciences, Beijing, China
Ming Li
University of Chinese Academy of Sciences, Beijing, China; Institute of Physics, Chinese Academy of Sciences, Beijing, China
BLM is a multifunctional helicase that plays critical roles in maintaining genome stability. It processes distinct DNA substrates, but not nicked DNA, during many steps in DNA replication and repair. However, how BLM prepares itself for diverse functions remains elusive. Here, using a combined single-molecule approach, we find that a high abundance of BLMs can indeed unidirectionally unwind dsDNA from a nick when an external destabilizing force is applied. Strikingly, human replication protein A (hRPA) not only ensures that limited quantities of BLMs processively unwind nicked dsDNA under a reduced force but also permits the translocation of BLMs on both intact and nicked ssDNAs, resulting in a bidirectional unwinding mode. This activation necessitates BLM targeting on the nick and the presence of free hRPAs in solution whereas direct interactions between them are dispensable. Our findings present novel DNA unwinding activities of BLM that potentially facilitate its function switching in DNA repair.
