An In Vitro Transcription System that Recapitulates Equine Infectious Anemia Virus Tat-Mediated Inhibition of Human Immunodeficiency Virus Type 1 Tat Activity Demonstrates a Role for Positive Transcription Elongation Factor b and Associated Proteins in the Mechanism of Tat Activation

Carlos Suñé; Aaron C Goldstrohm; Junmin Peng; David H Price; Mariano A Garcia-Blanco

doi:10.1006/viro.2000.0480

Back

An In Vitro Transcription System that Recapitulates Equine Infectious Anemia Virus Tat-Mediated Inhibition of Human Immunodeficiency Virus Type 1 Tat Activity Demonstrates a Role for Positive Transcription Elongation Factor b and Associated Proteins in the Mechanism of Tat Activation

Journal article

Open access

Peer reviewed

An In Vitro Transcription System that Recapitulates Equine Infectious Anemia Virus Tat-Mediated Inhibition of Human Immunodeficiency Virus Type 1 Tat Activity Demonstrates a Role for Positive Transcription Elongation Factor b and Associated Proteins in the Mechanism of Tat Activation

Carlos Suñé, Aaron C Goldstrohm, Junmin Peng, David H Price and Mariano A Garcia-Blanco

Virology (New York, N.Y.), Vol.274(2), pp.356-366

09/01/2000

DOI: 10.1006/viro.2000.0480

PMID: 10964778

Files and links (1)

url

https://doi.org/10.1006/viro.2000.0480View

Published (Version of record) Open Access

Abstract

Equine infectious anemia virus (EIAV) activates transcription via a Tat protein, a TAR element, and the equine elongation factor positive transcription elongation factor b (P-TEFb). In human cells, EIAV Tat (eTat) can inhibit the ability of human immunodeficiency virus type 1 (HIV-1) Tat (hTat) to activate transcription from the HIV-1 long terminal repeat, demonstrating that EIAV Tat can interact nonproductively with human P-TEFb. To study the mechanism of EIAV Tat and HIV-1 Tat activation, we developed an in vitro elongation assay that recapitulates EIAV Tat-mediated inhibition of HIV-1 Tat trans-activation. We found that eTat specifically inhibits activation of elongation by HIV-1 Tat while having no effect on basal transcription elongation. The competitive inhibition of hTat activation was reversed by an activity present in HeLa cell nuclear extracts, most likely a form of P-TEFb. Recombinant P-TEFb (cyclin T1 and CDK9) overcame the inhibition of transcription by eTat but in a nonspecific manner. EIAV Tat affinity chromatography was used to purify the activity present in nuclear extract that was capable of reversing eTat inhibition. We characterized the protein components of this activity, which include cyclin T1, CDK9, Tat-SF1, and at least three unidentified proteins. These data suggest that additional factors are involved in the mechanism of Tat activation.

Details

Title: Subtitle: An In Vitro Transcription System that Recapitulates Equine Infectious Anemia Virus Tat-Mediated Inhibition of Human Immunodeficiency Virus Type 1 Tat Activity Demonstrates a Role for Positive Transcription Elongation Factor b and Associated Proteins in the Mechanism of Tat Activation
Creators: Carlos Suñé - Departments of Genetics, Duke University Medical Center, Durham, North Carolina, 27710
Aaron C Goldstrohm - Microbiology, Duke University Medical Center, Durham, North Carolina, 27710
Junmin Peng - Department of Biochemistry, University of Iowa, Iowa City, Iowa, 52242
David H Price - Department of Biochemistry, University of Iowa, Iowa City, Iowa, 52242
Mariano A Garcia-Blanco - Departments of Genetics, Duke University Medical Center, Durham, North Carolina, 27710
Resource Type: Journal article
Publication Details: Virology (New York, N.Y.), Vol.274(2), pp.356-366
DOI: 10.1006/viro.2000.0480
PMID: 10964778
NLM abbreviation: Virology
ISSN: 0042-6822
eISSN: 1096-0341
Publisher: Elsevier Inc
Language: English
Date published: 09/01/2000
Academic Unit: Biochemistry and Molecular Biology
Record Identifier: 9984025256102771

Metrics

27 Record Views

14 Times Cited - Web of Science