This proposal aims to enhance our understanding of the molecular interactions of the complexes involved in transcription initiation by RNA polymerase II. The transcription of DMA into RNA is a universal mechanism that is required of all cells for proper gene expression. Irregular initiation of transcription can lead to a variety of cellular maladies, ranging from apoptosis and aberrant differentiation to unchecked proliferation and growth. The protein complexes involved in transcription initiation are highly conserved among eukaryotes. In addition to the core RNA polymerase enzyme, the transcription machinery is also composed of associated complexes known as general transcription factors, including TFIIA, TFIIB, TFIID, TFIIF, and TFIIH. Using Saccharomyces cerevisiae, a tractable organism that can be utilized to easily obtain large quantities of these factors, this project will biochemically dissect how the general transcription factors and RNA polymerase II interact to not only form the initiation complex, but to regulate the transition to an active and processive transcription complex as well. The transcription factors, along with the core polymerase, will be purified either in recombinant forms or from the endogenous source. Taking advantage of a variety of molecular techniques, the role each factor plays during the initiation phase will be determined as well as methods of regulation in order to give us further insights as to how gene expression is coordinated.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31GM075383-02
Application #
7123852
Study Section
Special Emphasis Panel (ZRG1-F08 (29))
Program Officer
Toliver, Adolphus
Project Start
2005-09-01
Project End
2008-08-31
Budget Start
2006-09-01
Budget End
2007-08-31
Support Year
2
Fiscal Year
2006
Total Cost
$28,580
Indirect Cost
Name
Harvard University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115
Dermody, Jessica L; Buratowski, Stephen (2010) Leo1 subunit of the yeast paf1 complex binds RNA and contributes to complex recruitment. J Biol Chem 285:33671-9
Dermody, Jessica L; Dreyfuss, Jonathan M; Villen, Judit et al. (2008) Unphosphorylated SR-like protein Npl3 stimulates RNA polymerase II elongation. PLoS One 3:e3273