Sequence-specific interaction of Tat protein and Tat peptides with the transactivation-responsive sequence element of human immunodeficiency virus type 1 in vitro.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 22 ·1990-11-00 ·Pages 8985-9

Cordingley MG, LaFemina RL, Callahan PL, Condra JH, Sardana VV, Graham DJ, Nguyen TM, LeGrow K, Gotlib L, Schlabach AJ

Abstract

Bacterially expressed Tat protein of human immunodeficiency virus type 1 binds selectively to short RNA transcripts containing the viral transactivation-responsive element (TAR). Sequences sufficient for Tat interaction map to the distal portion of the TAR stem-loop. We show that critical sequences for Tat binding are located in the single-stranded "bulge," but no requirement for specific "loop" sequences could be demonstrated. TAR RNA competed for complex formation, and TAR mutants exhibited up to 10-fold reduced affinity for Tat. Synthetic peptides containing the basic region of Tat bound selectively to TAR RNA and exhibited the same sequence requirements and similar relative affinities for mutant TAR RNA as the intact protein. These results suggest that Tat contains a small RNA-binding domain capable of recognizing TAR and implicate functional relevance for direct Tat-TAR interaction in transactivation.

MeSH Terms

Amino Acid Sequence Binding, Competitive DNA Mutational Analysis Gene Products, tat/metabolism HIV Long Terminal Repeat/genetics HIV-1/genetics In Vitro Techniques Molecular Sequence Data Peptides/chemical synthesis,metabolism RNA, Viral/genetics,metabolism Recombinant Proteins Regulatory Sequences, Nucleic Acid Structure-Activity Relationship tat Gene Products, Human Immunodeficiency Virus

Chemicals

Gene Products, tat Peptides RNA, Viral Recombinant Proteins tat Gene Products, Human Immunodeficiency Virus

Authors & Affiliations

10 authors, click to expand affiliations / ORCID

Cordingley M G

Department of Virus and Cell Biology, Merck Sharp and Dohme Research Laboratories, West Point, PA 19486.

LaFemina R L

Callahan P L

Condra J H

Sardana V V

Graham D J

Nguyen T M

LeGrow K

Gotlib L

Schlabach A J

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.87.22.8985

Published

1990-11-00

Pages

8985-9

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC55085

Subset

External Links

PubMed source DOI: 10.1073/pnas.87.22.8985 PMC Full Text View journal details

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Sequence-specific interaction of Tat protein and Tat peptides with the transactivation-responsive sequence element of human immunodeficiency virus type 1 in vitro.

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