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PMID: 9121429 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The human immunodeficiency virus transactivator Tat interacts with the RNA polymerase II holoenzyme.

Molecular and cellular biology ·Vol. 17 ·No. 4 ·1997-04-00 ·Pages 1817-23

Cujec TP, Cho H, Maldonado E, Meyer J, Reinberg D, Peterlin BM

Abstract

The human immunodeficiency virus (HIV) encodes a transcriptional transactivator (Tat), which binds to an RNA hairpin called the transactivation response element (TAR) that is located downstream of the site of initiation of viral transcription. Tat stimulates the production of full-length viral transcripts by RNA polymerase II (pol II). In this study, we demonstrate that Tat coimmunoprecipitates with the pol II holoenzyme in cells and that it binds to the purified holoenzyme in vitro. Furthermore, Tat affinity chromatography purifies a holoenzyme from HeLa nuclear extracts which, upon addition of TBP and TFIIB, supports Tat transactivation in vitro, indicating that it contains all the cellular proteins required for the function of Tat. By demonstrating that Tat interacts with the holoenzyme in the absence of TAR, our data suggest a single-step assembly of Tat and the transcription complex on the long terminal repeat of HIV.

MeSH Terms
Animals Base Sequence Binding Sites/genetics COS Cells DNA Primers/genetics Gene Products, tat/genetics,metabolism HIV Long Terminal Repeat HIV-1/genetics,metabolism Humans In Vitro Techniques Mutagenesis, Site-Directed Polymerase Chain Reaction RNA Polymerase II/metabolism Rats Transcription, Genetic tat Gene Products, Human Immunodeficiency Virus
Chemicals
DNA Primers Gene Products, tat tat Gene Products, Human Immunodeficiency Virus RNA Polymerase II
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Cujec T P
Department of Medicine, University of California at San Francisco, 94143-0724, USA.
Cho H
Maldonado E
Meyer J
Reinberg D
Peterlin B M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.4.1817
Published
1997-04-00
Pages
1817-23
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232028
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.17.4.1817 PMC Full Text View journal details
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