National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
AI094386
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM071940
米国
National Institutes of Health/National Institute of Dental and Craniofacial Research (NIH/NIDCR)
DE025567
米国
National Institutes of Health/National Center for Research Resources (NIH/NCRR)
1S10RR23057
米国
National Institutes of Health/Office of the Director
1S10OD018111
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
U24GM116792
米国
National Science Foundation (NSF, United States)
DBI-1338135
米国
National Science Foundation (NSF, United States)
DMR-1548924
米国
引用
ジャーナル: Nat Commun / 年: 2019 タイトル: In situ structures of rotavirus polymerase in action and mechanism of mRNA transcription and release. 著者: Ke Ding / Cristina C Celma / Xing Zhang / Thomas Chang / Wesley Shen / Ivo Atanasov / Polly Roy / Z Hong Zhou / 要旨: Transcribing and replicating a double-stranded genome require protein modules to unwind, transcribe/replicate nucleic acid substrates, and release products. Here we present in situ cryo-electron ...Transcribing and replicating a double-stranded genome require protein modules to unwind, transcribe/replicate nucleic acid substrates, and release products. Here we present in situ cryo-electron microscopy structures of rotavirus dsRNA-dependent RNA polymerase (RdRp) in two states pertaining to transcription. In addition to the previously discovered universal "hand-shaped" polymerase core domain shared by DNA polymerases and telomerases, our results show the function of N- and C-terminal domains of RdRp: the former opens the genome duplex to isolate the template strand; the latter splits the emerging template-transcript hybrid, guides genome reannealing to form a transcription bubble, and opens a capsid shell protein (CSP) to release the transcript. These two "helicase" domains also extensively interact with CSP, which has a switchable N-terminal helix that, like cellular transcriptional factors, either inhibits or promotes RdRp activity. The in situ structures of RdRp, CSP, and RNA in action inform mechanisms of not only transcription, but also replication.