Human immunodeficiency virus rev protein recognizes a target sequence in rev-responsive element RNA within the context of RNA secondary structure [published erratum appears in J Virol 1992 Feb;66(2):1288]

Important note: Information in this article was accurate in 1991. The state of the art may have changed since the publication date.

J Virol. 1990 Dec;64(12):5966-75. Unique Identifier : AIDSLINE MED/91056566
Holland SM; Ahmad N; Maitra RK; Wingfield P; Venkatesan S; Laboratory of Molecular Microbiology, National Institute of; Allergy and Infectious Diseases, National Institutes of Health,; Bethesda, Maryland 20892.

Abstract: Human immunodeficiency virus type 1 Rev protein modulates the distribution of viral mRNAs from the nucleus to the cytoplasm by interaction with a highly structured viral RNA sequence, the Rev-responsive element (RRE). To identify the minimal functional elements of RRE, we evaluated mutant RREs for Rev binding in vitro and Rev response in vivo in the context of a Gag expression plasmid. The critical functional elements fold into a structure composed of a stem-loop A, formed by the ends of the RRE, joined to a branched stem-loop B/B1/B2, between bases 49 and 113. The 5' 132 nucleotides of RRE, RREDDE, which possessed a similar structure, bound Rev efficiently but were nonfunctional in vivo, implying separate binding and functional domains within the RRE. Excision of stem-loop A reduced Rev binding significantly and abolished the in vivo Rev response. The B2 branch could be removed without severe impairment of binding, but deletions in the B1 branch significantly reduced binding and function. However, deletion of 12 nucleotides, including the 5' strand of stem B, abolished both binding and function, while excision of the 3' strand of stem B only reduced them. Maintenance of the native RRE secondary structure alone was not sufficient for Rev recognition. Many mutations that altered the primary structure of the critical region while preserving the original RNA conformation were Rev responsive. However, mutations that changed a 5'..CACUAUGGG..3' sequence in the B stem, without affecting the overall structure abolished both in vitro Rev binding and the in vivo Rev response.

Keywords: Base Sequence Chromosome Deletion Cloning, Molecular Escherichia coli/GENETICS Gene Products, rev/*METABOLISM Human HIV-1/*GENETICS/METABOLISM Kinetics Molecular Sequence Data Mutagenesis, Site-Directed Nucleic Acid Conformation Polymerase Chain Reaction Protein Binding Recombinant Proteins/METABOLISM Repetitive Sequences, Nucleic Acid Restriction Mapping RNA, Messenger/*GENETICS RNA, Viral/BIOSYNTHESIS/*GENETICS Support, U.S. Gov't, P.H.S. Transcription, Genetic Transfection JOURNAL ARTICLE
910330
M9130547

AEGiS is a 501(c)3, not-for-profit, tax-exempt, educational corporation. AEGiS is made possible through unrestricted funding from Boehringer Ingelheim, Bridgestone/Firestone Charitable Trust, Bristol-Myers Squibb Company, Elton John AIDS Foundation, Gill Foundation, the National Library of Medicine, Quest Diagnostics, Roche and Trimeris, and donations from users like you. Always watch for outdated information. This article first appeared in 1991. This material is designed to support, not replace, the relationship that exists between you and your doctor.

AEGiS presents published material, reprinted with permission and neither endorses nor opposes any material. All information contained on this website, including information relating to health conditions, products, and treatments, is for informational purposes only. It is often presented in summary or aggregate form. It is not meant to be a substitute for the advice provided by your own physician or other medical professionals. Always discuss treatment options with a doctor who specializes in treating HIV.

Copyright ©1980, 1991. AEGiS. All materials appearing on AEGiS are protected by copyright as a collective work or compilation under U.S. copyright and other laws and are the property of AEGiS, or the party credited as the provider of the content. .