Conservative transcription in three steps visualized in a double-stranded RNA virus

Endogenous RNA transcription characterizes double-stranded RNA (dsRNA) viruses in the Reoviridae, a family that is exemplified by its simple, single-shelled member cytoplasmic polyhedrosis virus (CPV). Because of the lack of in situ structures of the intermediate stages of RNA-dependent RNA polymera...

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Published inNature structural & molecular biology Vol. 26; no. 11; pp. 1023 - 1034
Main Authors Cui, Yanxiang, Zhang, Yinong, Zhou, Kang, Sun, Jingchen, Zhou, Z Hong
Format Journal Article
LanguageEnglish
Published United States Nature Publishing Group 01.11.2019
Subjects
Cryoelectron Microscopy
DNA-directed RNA polymerase
Double-stranded RNA
Elongated structure
Elongation
Genetic aspects
Genetic research
Genetic transcription
Humans
Models, Molecular
Reoviridae - chemistry
Reoviridae - genetics
Reoviridae - ultrastructure
Reoviridae Infections - virology
RNA polymerase
RNA polymerases
RNA viruses
RNA, Double-Stranded - chemistry
RNA, Double-Stranded - genetics
RNA, Double-Stranded - ultrastructure
RNA, Viral - chemistry
RNA, Viral - genetics
RNA, Viral - ultrastructure
RNA-Dependent RNA Polymerase - chemistry
RNA-Dependent RNA Polymerase - ultrastructure
RNA-directed RNA polymerase
Structure
Transcription elongation
Transcription, Genetic
Viral Proteins - chemistry
Viral Proteins - ultrastructure
Viruses
United States
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Summary:Endogenous RNA transcription characterizes double-stranded RNA (dsRNA) viruses in the Reoviridae, a family that is exemplified by its simple, single-shelled member cytoplasmic polyhedrosis virus (CPV). Because of the lack of in situ structures of the intermediate stages of RNA-dependent RNA polymerase (RdRp) during transcription, it is poorly understood how RdRp detects environmental cues and internal transcriptional states to initiate and coordinate repeated cycles of transcript production inside the capsid. Here, we captured five high-resolution (2.8-3.5 Å) RdRp-RNA in situ structures-representing quiescent, initiation, early elongation, elongation and abortive states-under seven experimental conditions of CPV. We observed the 'Y'-form initial RNA fork in the initiation state and the complete transcription bubble in the elongation state. These structures reveal that de novo RNA transcription involves three major conformational changes during state transitions. Our results support an ouroboros model for endogenous conservative transcription in dsRNA viruses.
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Z.H.Z. and J.S. conceived, designed and oversaw the project; J.S. and Y.Z. prepared and carried out experimental reactions; Y.C. prepared cryoEM samples, acquired cryoEM movies and performed data processing; Y.Z. built atomic models with assistance from K.Z.; Y.Z., Y.C. and Z.H.Z. interpreted the structures; Z.H.Z. and Y.Z. wrote the initial draft of the paper and all authors edited and approved the paper.
Author contributions
ISSN:1545-9993
1545-9985
DOI:10.1038/s41594-019-0320-0