Our long term goal is to elucidate the mechanisms of initiation and regulation of RNA polymerase II transcription. Our experimental approach is to reconstitute these processes with purified proteins from yeast, and bring the power of combined biochemical and genetic analysis to bear on the mechanisms. During the previous project period, we developed a fully defined, appropriately regulated transcription system. The sub cloning of genes for all components and expression of many of the proteins in recombinant form should enable the culmination of our efforts during the coming project period.
Specific aims of the proposed research are as follows: 1. Reconstitution and functional analysis of TFIIH. 2. Reconstitution and functional analysis of mediator. 3. Characterization of mammalian mediator. 4. Reconstitution of yeast TAF-dependent transcription in vitro. 5. Studies of RSC-nucleosome interaction.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37GM036659-17
Application #
6519222
Study Section
Molecular Biology Study Section (MBY)
Program Officer
Tompkins, Laurie
Project Start
1986-07-01
Project End
2003-06-30
Budget Start
2002-07-01
Budget End
2003-06-30
Support Year
17
Fiscal Year
2002
Total Cost
$464,044
Indirect Cost
Name
Stanford University
Department
Biology
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Lorch, Yahli; Maier-Davis, Barbara; Kornberg, Roger D (2018) Histone Acetylation Inhibits RSC and Stabilizes theĀ +1 Nucleosome. Mol Cell 72:594-600.e2
Nagai, Shigeki; Davis, Ralph E; Mattei, Pierre Jean et al. (2017) Chromatin potentiates transcription. Proc Natl Acad Sci U S A 114:1536-1541
Eagen, Kyle P; Aiden, Erez Lieberman; Kornberg, Roger D (2017) Polycomb-mediated chromatin loops revealed by a subkilobase-resolution chromatin interaction map. Proc Natl Acad Sci U S A 114:8764-8769
Robinson, Philip J; Trnka, Michael J; Bushnell, David A et al. (2016) Structure of a Complete Mediator-RNA Polymerase II Pre-Initiation Complex. Cell 166:1411-1422.e16
Lorch, Yahli; Kornberg, Roger D (2015) Chromatin-remodeling and the initiation of transcription. Q Rev Biophys 48:465-70
Guan, Shenheng; Trnka, Michael J; Bushnell, David A et al. (2015) Deconvolution method for specific and nonspecific binding of ligand to multiprotein complex by native mass spectrometry. Anal Chem 87:8541-6
Eagen, Kyle P; Hartl, Tom A; Kornberg, Roger D (2015) Stable Chromosome Condensation Revealed by Chromosome Conformation Capture. Cell 163:934-46
Fazal, Furqan M; Meng, Cong A; Murakami, Kenji et al. (2015) Real-time observation of the initiation of RNA polymerase II transcription. Nature 525:274-7
Lu, Jonathan; Trnka, Michael J; Roh, Soung-Hun et al. (2015) Improved Peak Detection and Deconvolution of Native Electrospray Mass Spectra from Large Protein Complexes. J Am Soc Mass Spectrom 26:2141-51
Murakami, Kenji; Mattei, Pierre-Jean; Davis, Ralph E et al. (2015) Uncoupling Promoter Opening from Start-Site Scanning. Mol Cell 59:133-8

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