The Rev-mediated Dimerization of HIV RRE RNA

dc.contributor.advisorEpstein, Henry
dc.contributor.committeeMemberFox, Robert O
dc.contributor.committeeMemberBriggs, James
dc.contributor.committeeMemberCloyd, Miles
dc.contributor.committeeMemberIwahara, Junji
dc.creatorAllison, Jason 1974-
dc.date.accessioned2016-11-14T15:22:26Z
dc.date.available2016-11-14T15:22:26Z
dc.date.created2012-08
dc.date.submittedAugust 2012
dc.date.updated2016-11-14T15:22:27Z
dc.description.abstractThe HIV-1 Regulator of Virion Expression (Rev) a regulatory protein which is critical for the late-phase development of the Human Immunodeficiency Virus-1 (HIV-1). During early-phase development of HIV-1, Rev accumulates in the cytoplasm and is transported into the nucleus through interactions with Importin-β via its Nuclear Localization Sequence (NLS). Following sufficient accumulation in the nucleus, Rev recognizes and assembles on the Rev Response Element (RRE), a 351nt region contained within singly and unspliced HIV RNAs. This ribonucleoprotein (RNP) complex allows for the export of these RNA transcripts encoding viral structural proteins and genomic RNA to the cytoplasm. Of particular interest is the interaction of Rev with the Rev Response Element (RRE) RNA. It is known that Rev initially binds to a structured Stem Loop-IIB on the RRE and subsequently assembles along flanking sequences of the RRE; however, details of this interaction are not completely understood. RRE-containing tRNAs were designed for biophysical studies of Rev:RRE complexes. The tRNA chimeras employ natural tRNA from human lysyl tRNA, bacterial initiator methionyl tRNA, and baker’s yeast (S. cerevisiae) phenylalanyl tRNA for use as scaffolds to express RRE sequences. Using size-exclusion chromatography (SEC), analytical ultracentrifugation (AUC), and small angle X-ray scattering (SAXS), I demonstrate the formation of a discrete Rev:RRE RNA complex. Data analysis presented in this work suggests that Rev is mediating a dimer of RRE RNA.
dc.format.mimetypeapplication/pdf
dc.identifier.urihttp://hdl.handle.net/2152.3/826
dc.subjectHIV Rev Protein
dc.subjectchimeric tRNA:stem loop IIB:Rev Response Element
dc.titleThe Rev-mediated Dimerization of HIV RRE RNA
dc.typeThesis
dc.type.materialtext
thesis.degree.departmentBiochemistry and Molecular Biology
thesis.degree.disciplineMolecular Biophysics
thesis.degree.grantorThe University of Texas Medical Branch at Galveston
thesis.degree.levelDoctoral
thesis.degree.nameMolecular Biophysics Educational Track (Doctoral)

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