Transcription initiation at a eukaryotic protein-encoding gene involves assembly of a multiprotein-DNA complex containing RNA polymerase II, SRB/Mediator components, six general transcription factors (GTFs), and promoter DNA. The fully assembled transcription initiation complex contains more than 50 distinct polypeptides and has a molecular mass of more than 3 MDa. The proposed work addresses the structure of the transcription initiation complex, the pathway of assembly of the complex, and the promoter sequence elements that govern stability, assembly, and function of the complex. The proposed work has four specific aims:
Specific Aim 1 : To use systematic site-specific protein-DNA photocrosslinking to map protein-DNA interactions within the transcription initiation complex.
Specific Aim 2 : To use kinetic site-specific protein-DNA photocrosslinking to define the kinetics of formation of protein-DNA interactions within the transcription initiation complex.
Specific Aim 3 : To use binding-site selection to define promoter sequence elements recognized by GTFs.
Specific Aim 4 : To use high-density-array expression analysis to define the functional significance of promoter sequence elements recognized by GTFs. The results to be obtained will contribute to understanding human transcription initiation, to understanding human transcription regulation, and to design and synthesis of low-molecular-weight inhibitors of protein-DNA interactions in human transcription initiation, for application in selective regulation of transcription initiation in vitro and in vivo, and possibly for application in selective antiviral and anticancer therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM053665-08
Application #
6636156
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Tompkins, Laurie
Project Start
1996-03-01
Project End
2005-02-28
Budget Start
2003-03-01
Budget End
2005-02-28
Support Year
8
Fiscal Year
2003
Total Cost
$172,926
Indirect Cost
Name
Rutgers University
Department
Type
Organized Research Units
DUNS #
001912864
City
New Brunswick
State
NJ
Country
United States
Zip Code
08901
Ho, Mary X; Hudson, Brian P; Das, Kalyan et al. (2009) Structures of RNA polymerase-antibiotic complexes. Curr Opin Struct Biol 19:715-23