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Title: Initiation of RNA Polymerization and Polymerase Encapsidation by a Small dsRNA Virus

Journal Article · · PLoS Pathogens
 [1];  [2];  [1];  [1];  [2];  [1]
  1. Rice Univ., Houston, TX (United States)
  2. Univ. of Helsinki (Finland)
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During the replication cycle of double-stranded (ds) RNA viruses, the viral RNA-dependent RNA polymerase (RdRP) replicates and transcribes the viral genome from within the viral capsid. How the RdRP molecules are packaged within the virion and how they function within the confines of an intact capsid are intriguing questions with answers that most likely vary across the different dsRNA virus families. In this study, we have determined a 2.4 Å resolution structure of an RdRP from the human picobirnavirus (hPBV). In addition to the conserved polymerase fold, the hPBV RdRP possesses a highly flexible 24 amino acid loop structure located near the C-terminus of the protein that is inserted into its active site. In vitro RNA polymerization assays and site-directed mutagenesis showed that: (1) the hPBV RdRP is fully active using both ssRNA and dsRNA templates; (2) the insertion loop likely functions as an assembly platform for the priming nucleotide to allow de novo initiation; (3) RNA transcription by the hPBV RdRP proceeds in a semi-conservative manner; and (4) the preference of virus-specific RNA during transcription is dictated by the lower melting temperature associated with the terminal sequences. Co-expression of the hPBV RdRP and the capsid protein (CP) indicated that, under the conditions used, the RdRP could not be incorporated into the recombinant capsids in the absence of the viral genome. Additionally, the hPBV RdRP exhibited higher affinity towards the conserved 5’-terminal sequence of the viral RNA, suggesting that the RdRP molecules may be encapsidated through their specific binding to the viral RNAs during assembly.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
Gulf Coast Consortia; Viikki Doctoral Programme in Molecular Biosciences; National Institutes of Health (NIH); Welch Foundation; Academy of Finland; Sigrid Jusélius Foundation
Grant/Contract Number:
AIO67638 to YJT; C-1565; 250113; 256069; 283192
OSTI ID:
1438860
Journal Information:
PLoS Pathogens, Vol. 12, Issue 4; ISSN 1553-7374
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 36 works
Citation information provided by
Web of Science

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Cited By (5)

RNA Dependent RNA Polymerases: Insights from Structure, Function and Evolution journal February 2018
In situ structures of rotavirus polymerase in action and mechanism of mRNA transcription and release journal May 2019
Synthesis and Translation of Viral mRNA in Reovirus-Infected Cells: Progress and Remaining Questions journal November 2018
Detection and Molecular Characterization of Picobirnaviruses (PBVs) in the Mongoose: Identification of a Novel PBV Using an Alternative Genetic Code journal January 2020
Dual Role of a Viral Polymerase in Viral Genome Replication and Particle Self-Assembly journal October 2018

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59 BASIC BIOLOGICAL SCIENCES
Polymerases
Viral packaging
RNA synthesis
dsRNA viruses
Viral structure
Sequence motif analysis
RNA structure
Viral genomics