DOI: 10.1101/gad.11.20.2633. P-TEFb kinase is required for HIV Tat transcriptional activation in vivo and in vitro

P-TEFb kinase is required for HIV Tat transcriptional activation in vivo and in vitro

10.1101/gad.11.20.2633

Crossref journal-article

Cold Spring Harbor Laboratory

Genes & Development (246)

Abstract

To identify novel inhibitors of transcriptional activation by the HIV Tat protein, we used a combination of in vitro and in vivo Tat-dependent transcription assays to screen >100,000 compounds. All compounds identified blocked Tat-dependent stimulation of transcriptional elongation. Analysis of a panel of structurally diverse inhibitors indicated that their target is the human homolog ofDrosophila positive transcription elongation factor b (P-TEFb). Loss of Tat transactivation in extracts depleted of the kinase subunit of human P-TEFb, PITALRE, was reversed by addition of partially purified human P-TEFb. Transfection experiments with wild-type or kinase knockout PITALRE demonstrated that P-TEFb is required for Tat function. Our results suggest that P-TEFb represents an attractive target for the development of novel HIV therapeutics.

Bibliography

Mancebo, H. S. Y., Lee, G., Flygare, J., Tomassini, J., Luu, P., Zhu, Y., Peng, J., Blau, C., Hazuda, D., Price, D., & Flores, O. (1997). P-TEFb kinase is required for HIV Tat transcriptional activation in vivo and inâvitro. Genes & Development, 11(20), 2633â2644.

Authors 11

Helena S.Y. Mancebo (first)
Gary Lee (additional)
John Flygare (additional)
Joanne Tomassini (additional)
Percy Luu (additional)
Yuerong Zhu (additional)
Junmin Peng (additional)
Carol Blau (additional)
Daria Hazuda (additional)
David Price (additional)
Osvaldo Flores (additional)

References 45 Referenced 448

10.1038/377557a0
{'key': '2021111418475564000_11.20.2633.2', 'first-page': '4.7.1', 'article-title': 'RNase protection assay.', 'volume': '1', 'author': 'Ausubel', 'year': '1995', 'journal-title': 'Current protocols in molecular biology.'} / Current protocols in molecular biology. / RNase protection assay. by Ausubel (1995)
10.1128/MCB.16.5.2044 / Mol. Cell. Biol. / Three functional classes of transcriptional activation domains. by Blau (1996)
10.1016/S0076-6879(96)75021-8 / Methods Enzymol. / A system to analyze and identify inhibitors of HIV-1 gene regulation using a defective integrated provirus. by Byrnes (1996)
10.1016/S0021-9258(20)80751-0 / J. Biol. Chem. / RAP30/74 (transcription factor IIF) is required for promoter escape by RNA polymerase II. by Chang (1993)
10.1016/S0021-9258(18)94169-4 / J. Biol. Chem. / 5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole inhibits transcription elongation by RNA polymerase II in vitro. by Chodosh (1989)
10.1074/jbc.271.32.19009
10.1073/pnas.88.9.4045
10.1016/S0021-9258(18)45947-9 / J. Biol. Chem. / Factors involved in specific transcription by mammalian RNA polymerase II. Identification and characterization of factor IIH. by Flores (1992)
10.1073/pnas.76.6.2571
10.1093/emboj/16.10.2836
10.1016/0092-8674(94)90242-9
10.1038/352689a0
10.1006/viro.1993.1634
10.1128/jvi.69.3.1612-1620.1995 / J. Virol. / Lentivirus Tat proteins specifically associate with a cellular protein kinase, TAK, that hyperphosphorylates the carboxyl-terminal domain of the large subunit of RNA polymerase II: Candidate for a Tat cofactor. (1995)
10.1042/bst0200525
10.1146/annurev.bi.63.070194.003441
{'key': '2021111418475564000_11.20.2633.18', 'first-page': '1238', 'article-title': 'In vivo and in vitro analysis of transcriptional activation mediated by the human cytomegalovirus major immediate-early proteins.', 'volume': '13', 'author': 'Klucher', 'year': '1993', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / In vivo and in vitro analysis of transcriptional activation mediated by the human cytomegalovirus major immediate-early proteins. by Klucher (1993)
10.1101/gad.9.5.559
10.1016/0092-8674(89)90290-0
10.1101/gad.4.12b.2397
10.1073/pnas.77.6.3297
10.1073/pnas.93.6.2570
10.1073/pnas.88.22.10004
10.1002/j.1460-2075.1991.tb04997.x / EMBO J. / HIV-1 Tat protein promotes formation of more-processive elongation complexes. by Marciniak (1991)
10.1016/0092-8674(90)90145-5
10.1074/jbc.271.43.27176
10.1021/jm00017a003
10.1128/jvi.64.9.4565-4567.1990 / J. Virol. / Human chromosome 12 encodes a species-specific factor which increases human immunodeficiency virus type 1 Tat-mediated transactivation in rodent cells. by Newstein (1990)
10.1038/384375a0
10.1038/354369a0
10.1016/S0021-9258(18)61506-6 / J. Biol. Chem. / Factors involved in specific transcription in mammalian RNA polymerase II. Functional analysis of initiation factors IIA and IID and identification of a new factor operating at sequences downstream of the initiation site. by Reinberg (1987)
10.1101/gad.5.12b.2508
10.1073/pnas.76.11.5750
10.1093/nar/9.14.3307
10.1128/jvi.67.4.2349-2354.1993 / J. Virol. / The blocks to human immunodeficiency virus type 1 Tat and Rev functions in mouse cell lines are independent. by Winslow (1993)
10.1128/jvi.70.7.4576-4584.1996 / J. Virol. / The human immunodeficiency virus Tat proteins specifically associate with TAK in vivo and require the carboxyl-terminal domain of RNA polymerase II for function. by Yang (1996)
10.1016/0092-8674(94)90058-2
10.1128/MCB.16.7.3291 / Mol. Cell. Biol. / TFIIH functions in regulating transcriptional elongation by RNA polymerase II in Xenopus oocytes. by Yankulov (1996)
10.1016/S0021-9258(17)42674-3 / J. Biol. Chem. / Inhibitory effect of 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole on a protein kinase. by Zandomeni (1984)
10.1016/S0021-9258(17)35799-X / J. Biol. Chem. / Casein kinase type II is involved in the inhibition by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole of specific RNA polymerase II transcription. by Zandomeni (1986)
10.1002/j.1460-2075.1995.tb07006.x / EMBO J. / Novel mechanism and factor for regulation by HIV-1 Tat. by Zhou (1995)
10.1126/science.274.5287.605
Zhu, Y., T. Pe’ery, J. Peng, Y. Ramanathan, N. Marshall, T. Marshall, B. Amendt, M.B. Mathews, and D.H. Price. 1997. Transcription elongation factor P-TEFb is required for HIV-1 Tat transactivation in vitro.Genes & Dev. (this issue).. (10.1101/gad.11.20.2622)
10.1021/jm960157v

Dates

Type	When
Created	17 years, 6 months ago (Feb. 20, 2008, 5:15 p.m.)
Deposited	3 years, 9 months ago (Nov. 14, 2021, 10:10 p.m.)
Indexed	2 weeks ago (Aug. 19, 2025, 6:22 a.m.)
Issued	27 years, 10 months ago (Oct. 15, 1997)
Published	27 years, 10 months ago (Oct. 15, 1997)
Published Online	27 years, 10 months ago (Oct. 15, 1997)
Published Print	27 years, 10 months ago (Oct. 15, 1997)

Funders 0

None

BibTeX

@article{Mancebo_1997, title={P-TEFb kinase is required for HIV Tat transcriptional activation in vivo and in vitro}, volume={11}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.11.20.2633}, DOI={10.1101/gad.11.20.2633}, number={20}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={Mancebo, Helena S.Y. and Lee, Gary and Flygare, John and Tomassini, Joanne and Luu, Percy and Zhu, Yuerong and Peng, Junmin and Blau, Carol and Hazuda, Daria and Price, David and Flores, Osvaldo}, year={1997}, month=oct, pages={2633–2644} }

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