18. GOALS FOR FELLOWSHIP TRAINING AND CAREER Training goals under this fellowship are two-tiered. The first consists of tile development of analytical paradigms required to organize a successful research project. The second consists of development of specific subject knowledge and technical expertise related to eukaryotic transcription analysis culminating in the better understanding of eukaryotic transcription and cell cycle regulation. My intermediate- to long-term career goal is to obtain a faculty position at a research university. This goal will require not only a well-developed ability to design project proposals and goals but also the technical expertise to predict the outcomes and troubleshoot problems encountered during the research. The opportunity to gain advanced biochemical training will complement the molecular biological training in RNA processing obtained during my graduate work and should allow me to synergistically combine the approaches to study gene expression mechanisms. 19. NAME AND DEGREE(S) Brian D. D__ynlacht '_, POSffION/RANK Associate Professor RESEARCH INTERESTS/AREAS Mechanisms of transcriptional regulation by members of the pRB and E2F families of proteins 22. DESCRIPTION (Do not exceed space provided) The global objective of the proposed research is to biochemically characterize the chrolnatin-specific mechanisms by which lnembers 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. PHS 416-1 (Rev, 12/98) Form Page 2 BB cc Individual NRSA Application NAME (Last, first, middle initial) Table of Contents ========================================Section End===========================================
| 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 |
