FUNCTIONAL ANALYSIS OF THE TAT AND TRS GENES OF HUMAN IMMUNODEFICIENCY VIRUS BY SITE-DIRECTED MUTAGENESIS

Important note: Information in this article was accurate in 1989. The state of the art may have changed since the publication date.

FUNCTIONAL ANALYSIS OF THE TAT AND TRS GENES OF HUMAN IMMUNODEFICIENCY VIRUS BY SITE-DIRECTED MUTAGENESIS

Vaccines 88. New Chemical and Genetic Approaches to Vaccination: Prevention of AIDS and Other Viral, Bacterial, and Parasitic Diseases. Ginsberg H et al, eds. New York, Cold Spring Harbor Laboratory, p. 303-10, 1988.. Unique Identifier : AIDSLINE ICDB/89649773
Sadaie MR; Josephs SF; Rappaport J; Willis R; Gallo RC; Wong-Staal F; Benter T; Lab. of Tumor Cell Biology, NCI, Bethesda, MD 20892

Abstract: Missense mutants were constructed using a subclone of HIV containing the tat- and trs-coding regions; the subclone was then substituted into the parent wild-type vector, HXB2gpt. To generate the mutants, site-directed mutagenesis was targeted to the three major structural motifs of the functional first coding region of the tat gene: the amino-terminal proline-rich region, the middle cysteine-rich region, and the carboxy-terminal basic amino acid region. Alterations in the last of these regions affected the trs-coding sequence as well. The mutants were examined for tat activity, mRNA expression of wild-type and mutant proviruses was compared, and the effect of mutations at the level of protein and virus expression was determined. The tat protein trans-activates viral transcription, whereas the trs protein has an opposing effect. trs also is required for the appearance of stable mRNAs coding for structural proteins. Missense mutations within the putative metal-binding finger of tat revealed that this region is crucial for virus replication. One additional mutant outside this region had little effect on transcriptional activation; however, virus replication was abolished due to the alteration of a newly identified splice-acceptor site. The present data define critical sites necessary for tat/trs function and provide a basis for future studies directed toward the design of antiviral agents that would interrupt the trans-activation pathway. (8 Refs)

Keywords: Amino Acid Sequence Cloning, Molecular Gene Expression Regulation *Genes, Viral Human HIV/*GENETICS Molecular Sequence Data *Mutation Proviruses/GENETICS RNA, Messenger/GENETICS Transcription Factors/*GENETICS Transcription, Genetic Viral Proteins/*GENETICS MEETING PAPER

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