The Rev-mediated Dimerization of HIV RRE RNA

The 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.

Name thesis:degree_name

Molecular Biophysics Educational Track (Doctoral)

Level thesis:degree_level

Doctoral

Discipline thesis:degree_discipline

Molecular Biophysics

Grantor

The University of Texas Medical Branch at Galveston

Author dc:creator

• Allison, Jason 1974-

Advisor dc:contributor.advisor

• Epstein, Henry

Committee members dc:contributor.committeemember

• Fox, Robert O
• Briggs, James
• Cloyd, Miles
• Iwahara, Junji

• HIV Rev Protein
• chimeric tRNA:stem loop IIB:Rev Response Element

Handle dc:identifier.uri

http://hdl.handle.net/2152.3/826

OAI identifier oai:identifier

oai:utmb-ir.tdl.org:2152.3/826