The global objective of the proposed research is to biochemically characterize the chromatin-specific mechanisms by which members of the pRB family repress transcription, pRB has been shown in vitro and in vivo to function in the repression of many genes required for cell cycle progression. Indeed, mutations in the RB gene occur frequently in many kinds of cancer, implicating its role as a global regulator of cell division and differentiation. Available evidence suggests pRB functions by at least two distinct mechanisms. In the first, pRB binds to E2F and prevents the subsequent recruitment of TFIID and TFIIA, thus preventing formation of a pre-initiation complex (PIC). Recent work has shown that pRB functions by another E2F-independent mechanism dubbed """"""""active"""""""" repression that appears to involve the repression of chromatin remodeling. To gain a better understanding of this mechanism the following aims will be undertaken: (1) analysis of chromatin-modifying and -remodeling mechanisms achieved through E2F, Sp 1, and pRB family members on synthetic promoters in vitro, (2) analysis of regulation of mouse E2fl promoter in vitro by E2F, Spl, and pRB family members, and (3) study contribution of mechanisms characterized in Aims 1 and 2 to mouse E2fl promoter in living cells. In vitro transcriptional analysis, Chromatin Immunoprecipitation (ChIP) analysis, and luminescence based transcriptional analysis will be the main technical methods utilized in these aims in order to bridge basic biochemical analysis of transcription with physiologically relevant mechanisms controlling cell cycle progression.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
1F32GM064971-01A1
Application #
6584321
Study Section
Special Emphasis Panel (ZRG1-F05 (20))
Program Officer
Tompkins, Laurie
Project Start
2003-07-02
Project End
Budget Start
2003-02-01
Budget End
2004-01-31
Support Year
1
Fiscal Year
2003
Total Cost
$46,420
Indirect Cost
Name
Harvard University
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
082359691
City
Cambridge
State
MA
Country
United States
Zip Code
02138
Ciaccio, Mark F; Chen, Vincent C; Jones, Richard B et al. (2015) The DIONESUS algorithm provides scalable and accurate reconstruction of dynamic phosphoproteomic networks to reveal new drug targets. Integr Biol (Camb) 7:776-91