title: Structural Insight into the Specific DNA Template Binding to DnaG primase in Bacteria.
abstract: Bacterial primase initiates the repeated synthesis of short RNA primers that are extended by DNA polymerase to synthesize Okazaki fragments on the lagging strand at replication forks. It remains unclear how the enzyme recognizes specific initiation sites. In this study, the DnaG primase from Bacillus subtilis (BsuDnaG) was characterized and the crystal structure of the RNA polymerase domain (RPD) was determined. Structural comparisons revealed that the tethered zinc binding domain plays an important role in the interactions between primase and specific template sequence. Structural and biochemical data defined the ssDNA template binding surface as an L shape, and a model for the template ssDNA binding to primase is proposed. The flexibility of the DnaG primases from B. subtilis and G. stearothermophilus were compared, and the results implied that the intrinsic flexibility of the primase may facilitate the interactions between primase and various partners in the replisome. These results shed light on the mechanism by which DnaG recognizes the specific initiation site.
authors.affiliation: Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China.|Key Laboratory of Environmental Microbiology of Sichuan Province, Chengdu, 610041, China.|University of Chinese Academy of Sciences, Beijing, 100049, China.
authors.forename: Yingqin
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China.|Key Laboratory of Environmental Microbiology of Sichuan Province, Chengdu, 610041, China.|University of Chinese Academy of Sciences, Beijing, 100049, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Luo
authors.affiliation: Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China.|Key Laboratory of Environmental Microbiology of Sichuan Province, Chengdu, 610041, China.
authors.forename: Zhongchuan
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China.|Key Laboratory of Environmental Microbiology of Sichuan Province, Chengdu, 610041, China.|University of Chinese Academy of Sciences, Beijing, 100049, China.
authors.forename: Mu
authors.initials: M
authors.lastname: Yang
authors.affiliation: National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Pang
authors.affiliation: National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
authors.forename: Fei
authors.identifier: http://orcid.org/0000-0002-0351-5144
authors.initials: F
authors.lastname: Sun
authors.affiliation: Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China. wanggg@cib.ac.cn.|Key Laboratory of Environmental Microbiology of Sichuan Province, Chengdu, 610041, China. wanggg@cib.ac.cn.
authors.forename: Ganggang
authors.initials: G
authors.lastname: Wang
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism; D001419:Bacteria / Q000235:genetics* / Q000378:metabolism*; D001665:Binding Sites; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D019915:DNA Primase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D011485:Protein Binding; D059748:Proteolysis; D013329:Structure-Activity Relationship
chemical_list: D004247:DNA; D019915:DNA Primase