Initiation of RNA polymerization and polymerase encapsidation by a Picobirnavirus

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dc.contributor.advisorTao, Yizhi Jane
dc.contributor.committeeMemberStewart, Charles R
dc.contributor.committeeMemberOlson, John S
dc.contributor.committeeMemberPrasad, B.V. V
dc.creatorCollier, Aaron Michael
dc.date.accessioned2016-01-07T17:23:01Z
dc.date.available2016-01-07T17:23:01Z
dc.date.created2014-12
dc.date.issued2015-01-15
dc.date.submittedDecember 2014
dc.date.updated2016-01-07T17:23:01Z
dc.description.abstractDuring 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 these RdRPs molecules are packaged within the virion and how they function within the confines of an intact capsid are intriguing questions that have highly variable answers depending on the virus family being examined. In this study, we have determined a 2.4 Å resolution structure of an RdRP from a human infecting strain of picobirnavirus (PBV). In addition to a conserved polymerase fold, the PBV RdRP possesses a unique, highly flexible 24-aa loop structure (aa495-518) located near the C-terminus of the protein that is inserted into its active site. In vitro RNA polymerization assays have shown that the wild-type RdRP is capable of initiating RNA synthesis using a de novo mechanism, while a mutant RdRP lacking the loop structure could only synthesize RNA through back-priming, suggesting that the loop likely functions as a platform for the priming nucleotide to bind. Unexpectedly, co-expression of the PBV RdRP with its respective capsid protein (CP) indicated that the PBV RdRP could not be incorporated into recombinant capsids in the absence of the viral genome. Additionally, the PBV RdRP exhibited a high affinity towards the conserved 5’-terminal sequence of the viral genome, suggesting that PBV RdRP molecules are likely packaged through their specific binding to viral RNA during assembly.
dc.format.mimetypeapplication/pdf
dc.identifier.citationCollier, Aaron Michael. "Initiation of RNA polymerization and polymerase encapsidation by a Picobirnavirus." (2015) Diss., Rice University. <a href="https://hdl.handle.net/1911/87757">https://hdl.handle.net/1911/87757</a>.
dc.identifier.urihttps://hdl.handle.net/1911/87757
dc.language.isoeng
dc.rightsCopyright is held by the author, unless otherwise indicated. Permission to reuse, publish, or reproduce the work beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder.
dc.subjectdouble-stranded RNA virus
dc.subjectRNA-dependent RNA polymerase
dc.subjectvirus assembly
dc.subjectpicobirnavirus
dc.subjectgastroenteritis
dc.subjectRNA replication
dc.subjectRNA transcription
dc.subjectinitiation of RNA polymerization
dc.subjectback-priming
dc.subjectterminal nucleotidyl transferase
dc.titleInitiation of RNA polymerization and polymerase encapsidation by a Picobirnavirus
dc.typeThesis
dc.type.materialText
thesis.degree.departmentBiochemistry and Cell Biology
thesis.degree.disciplineNatural Sciences
thesis.degree.grantorRice University
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy
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