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PMID: 7729429 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Absolute dependence on kappa B responsive elements for initiation and Tat-mediated amplification of HIV transcription in blood CD4 T lymphocytes.

The EMBO journal ·Vol. 14 ·No. 7 ·1995-04-03 ·Pages 1552-60

Alcamí J, Laín de Lera T, Folgueira L, Pedraza MA, Jacqué JM, Bachelerie F, Noriega AR, Hay RT, Harrich D, Gaynor RB

Abstract

The role of NF-kappa B-dependent signals in activating the transcriptional activity of the HIV regulatory region (LTR) was analyzed by systematic comparison of HIV LTR activity in human CD4 T cells purified from peripheral blood and a transformed lymphoblastoid T cell line. In normal CD4 T cells we also analyzed the role played by the viral kappa B responsive elements in HIV replication. Analysis of nuclear extracts of resting, normal T lymphocytes revealed the presence of the p50, but not the p65, NF-kappa B subunit and the induction by phorbol esters of bona fide (p50-p65) NF-kappa B complexes. In parallel, we observed clear enhancer-dependent HIV LTR transactivation comparable in intensity with that observed in lymphoblastoid cells. We show that unstimulated CD4 T lymphocytes offer a cellular environment of very low permissivity to HIV LTR functioning. This was in sharp contrast to the high spontaneous LTR activity observed in lymphoblastoid T cells, where LTR activity was essentially independent of kappa B-responsive elements. Due to the low basal LTR activity in resting T lymphocytes, NF-kappa B-dependent transactivation was a sine qua non event for induction of the HIV LTR. Surprisingly, even the function of HIV Tat in resting CD4 T lymphocytes was found to be absolutely dependent on LTR kappa B responsive elements. The relevance of these observations obtained in transient transfections was confirmed by the incapacity of blood CD4 T lymphocytes infected with an HIV infectious provirus carrying critical point mutations in the kappa B responsive elements to show any detectable transcriptional activity upon cell activation and prolonged culture in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)

MeSH Terms
Base Sequence CD4-Positive T-Lymphocytes/physiology,virology Cell Line, Transformed Cells, Cultured DNA, Viral/biosynthesis Enhancer Elements, Genetic Gene Expression Regulation, Viral Gene Products, tat/biosynthesis,metabolism HIV/genetics,metabolism Humans Kinetics Molecular Sequence Data NF-kappa B/metabolism Proviruses/genetics,physiology RNA, Messenger/biosynthesis RNA, Viral/biosynthesis Signal Transduction T-Lymphocytes/physiology,virology Transcription, Genetic Transfection tat Gene Products, Human Immunodeficiency Virus
Chemicals
DNA, Viral Gene Products, tat NF-kappa B RNA, Messenger RNA, Viral tat Gene Products, Human Immunodeficiency Virus
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Alcamí J
Servicio de Microbiología, Hospital 12 de Octubre, Madrid, Spain.
Laín de Lera T
Folgueira L
Pedraza M A
Jacqué J M
Bachelerie F
Noriega A R
Hay R T
Harrich D
Gaynor R B
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-04-03
Pages
1552-60
Language
English
Region
England
NLM ID
8208664
PMCID
PMC398242
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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