Abstract

The objective of the application is to investigate the molecular mechanisms that underlie Tat and Rev regulation of HIV expression. The Sharp laboratory has recently identified a cellular factor, Tat-SF, that is required for Tat function. It was purified biochemically, using its intrinsic activity to monitor purifications. It has been microsequenced and the corresponding cDNA cloned. Tat-SF appears to assist in processivity of elongation by RNAP II in the presence of Tat, and it appears to interact with both Tat and TAR. The application proposes to further characterize Tat-SF-Tat interaction, and the role of these proteins in the processiveness of transcription in vitro and in vivo. The kinase that phosphorylates Tat-SF, and the components that further increase Tat-responsiveness in vitro will be further purified. The laboratory has also shown that the pol II elongation factor SIII increases polymerase processivity, and reduces Tat activation, suggesting that Tat may function through facilitation in the activity of SIII. The role of this factor in Tat activation will be further examined. Finally, the application also proposes to further study Rev regulation of RNA splicing, and the interaction of Rev with the nucleoporin protein Rab.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI032486-06
Application #
2003769
Study Section
AIDS and Related Research Study Section 3 (ARRC)
Project Start
1992-02-01
Project End
2002-01-31
Budget Start
1997-02-01
Budget End
1998-01-31
Support Year
6
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
Massachusetts Institute of Technology
Department
Type
Organized Research Units
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139

  Publications

Stewart, Sheila A; Dykxhoorn, Derek M; Palliser, Deborah et al. (2003) Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 9:493-501
Ge, Qing; McManus, Michael T; Nguyen, Tam et al. (2003) RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription. Proc Natl Acad Sci U S A 100:2718-23
Lee, Keng-Boon; Wang, Dong; Lippard, Stephen J et al. (2002) Transcription-coupled and DNA damage-dependent ubiquitination of RNA polymerase II in vitro. Proc Natl Acad Sci U S A 99:4239-44
Tuschl, T; Sharp, P A; Bartel, D P (2001) A ribozyme selected from variants of U6 snRNA promotes 2',5'-branch formation. RNA 7:29-43
Kim, J B; Sharp, P A (2001) Positive transcription elongation factor B phosphorylates hSPT5 and RNA polymerase II carboxyl-terminal domain independently of cyclin-dependent kinase-activating kinase. J Biol Chem 276:12317-23
Blencowe, B J; Bauren, G; Eldridge, A G et al. (2000) The SRm160/300 splicing coactivator subunits. RNA 6:111-20
Gilbert, S L; Sharp, P A (1999) Promoter-specific hypoacetylation of X-inactivated genes. Proc Natl Acad Sci U S A 96:13825-30
Mitsui, A; Sharp, P A (1999) Ubiquitination of RNA polymerase II large subunit signaled by phosphorylation of carboxyl-terminal domain. Proc Natl Acad Sci U S A 96:6054-9
Page-McCaw, P S; Amonlirdviman, K; Sharp, P A (1999) PUF60: a novel U2AF65-related splicing activity. RNA 5:1548-60
Kim, J B; Yamaguchi, Y; Wada, T et al. (1999) Tat-SF1 protein associates with RAP30 and human SPT5 proteins. Mol Cell Biol 19:5960-8

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