The human sequence-specific transcription factor USF is utilized by adenovirus for expression of the major late transcription unit. We have purified two different forms of this transcription factor from HeLa cell nuclei and isolated cDNA clones corresponding to one of these forms. These studies have revealed that the expression of this transcription factor, as well as its activity, may be subject to multiple levels of control. We now propose to isolate cDNA and antibody probes to the other form of USF and carry out a series of experiments which have two major objectives. The first objective will be to determine the cellular function of the USF proteins by investigating where and when these transcription factors are expressed and how their expression is regulated. Second, we would like to gain a better understanding of the molecular mechanisms by which USF stimulates transcription initiation by RNA polymerase II. For this, we propose to: i) dissect within these proteins the various domains which are involved in specific DNA binding, trans-activating function, or interaction with the TATA box factor TFIID and ii) analyze in detail the involvement of USF in the various steps of the transcription reaction, using a combination of physical (footprinting) as well as functional (template challenge) methods of analysis. In addition, we propose to investigate whether the possible interaction of USF with some components of the transcriptional machinery, or with other regulatory proteins, can be unraveled by using the bacterially-produced transcription factor as an affinity matrix.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM038212-07
Application #
3294384
Study Section
Molecular Biology Study Section (MBY)
Project Start
1987-04-01
Project End
1994-11-30
Budget Start
1992-12-01
Budget End
1993-11-30
Support Year
7
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Texas MD Anderson Cancer Center
Department
Type
Other Domestic Higher Education
DUNS #
001910777
City
Houston
State
TX
Country
United States
Zip Code
77030
Luo, X; Sawadogo, M (1996) Functional domains of the transcription factor USF2: atypical nuclear localization signals and context-dependent transcriptional activation domains. Mol Cell Biol 16:1367-75
Luo, X; Sawadogo, M (1996) Antiproliferative properties of the USF family of helix-loop-helix transcription factors. Proc Natl Acad Sci U S A 93:1308-13
Sawadogo, M; Van Dyke, M W (1995) Indirect use of immobilized metal affinity chromatography for isolation and characterization of protein partners. Genet Eng (N Y) 17:53-65
Szentirmay, M N; Sawadogo, M (1994) Sarkosyl block of transcription reinitiation by RNA polymerase II as visualized by the colliding polymerases reinitiation assay. Nucleic Acids Res 22:5341-6
Lin, Q; Luo, X; Sawadogo, M (1994) Archaic structure of the gene encoding transcription factor USF. J Biol Chem 269:23894-903
Sirito, M; Lin, Q; Maity, T et al. (1994) Ubiquitous expression of the 43- and 44-kDa forms of transcription factor USF in mammalian cells. Nucleic Acids Res 22:427-33
Lu, T; Sawadogo, M (1994) Role of the leucine zipper in the kinetics of DNA binding by transcription factor USF. J Biol Chem 269:30694-700
Meier, J L; Luo, X; Sawadogo, M et al. (1994) The cellular transcription factor USF cooperates with varicella-zoster virus immediate-early protein 62 to symmetrically activate a bidirectional viral promoter. Mol Cell Biol 14:6896-906
Szentirmay, M N; Sawadogo, M (1993) Synthesis of reinitiated transcripts by mammalian RNA polymerase II is controlled by elongation factor SII. EMBO J 12:4677-84
Lu, T; Van Dyke, M; Sawadogo, M (1993) Protein-protein interaction studies using immobilized oligohistidine fusion proteins. Anal Biochem 213:318-22

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