The long-term aim of this project is to elucidate the activity of Drosophila Retinoblastoma family (Rbf) proteins and associated cofactors in a developmental setting. Rbf proteins are key regulators of cell cycle control and play pivotal, although poorly understood, roles in development. Rbf proteins are differentially deployed during development and are thought to have distinct functions and effects on gene expression. The functional properties of Rbf proteins are dictated by the recruiting of distinct cofactors, and we have identified a novel association between the COP9 signalosome complex and Rbf2. Our preliminary data indicates that the COP9 signalosome may function in two important ways to influence Rbf function. First, the COP9 complex regulates Rbf protein stability in the developing embryo. Second, the COP9 complex may serve as a transcriptional co-repressor to enact patterns of transcriptional repression by Rbf proteins. This project will employ powerful biochemical and genetic tools that are available in Drosophila to develop a comprehensive picture of the COP9 signalosome in target gene regulation by Drosophila Rbf proteins. First, we will perform a detailed examination of the mechanism defining the association between the COP9 signalosome and Rbf proteins, as a vital pre-requisite to our subsequent functional assays. Second, we will examine the physiological role of the Rbf-associated COP9 complex using chromatin immunoprecipitation to determine when and where this putative cofactor is associated with Rbf proteins at target gene promoters. Third, the significance of Rbf protein instability and the role for the COP9 complex in this process during developmentally controlled gene repression patterns will be examined. Fourth, we will determine the physiological role of the COP9 signalosome as transcriptional co-regulatory factor for Rbf- mediated gene repression. The function of the COP9 complex as a co-repressor will be examined by means of genetic assays that test Rbf transcriptional regulatory activity during development and by transcription assays to determine the importance of individual COP9 subunits in Rbf regulation of specific genes in the fly. Retinoblastoma proteins have critical roles in development and are key regulators mutated in a high percentage of human tumors. Elucidation of the previously unidentified COP9 function for Rbf gene regulation, its mode of action, and its role in developmental gene regulation will provide important insight into RB action that will facilitate the design of therapeutic interventions in a wide spectrum of human cancers.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM079098-04
Application #
7777838
Study Section
Cellular Signaling and Dynamics Study Section (CSD)
Program Officer
Tompkins, Laurie
Project Start
2007-04-01
Project End
2011-03-31
Budget Start
2010-04-01
Budget End
2011-03-31
Support Year
4
Fiscal Year
2010
Total Cost
$275,881
Indirect Cost
Name
Michigan State University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
193247145
City
East Lansing
State
MI
Country
United States
Zip Code
48824
Payankaulam, Sandhya; Yeung, Kelvin; McNeill, Helen et al. (2016) Regulation of cell polarity determinants by the Retinoblastoma tumor suppressor protein. Sci Rep 6:22879
Wei, Yiliang; Mondal, Shamba S; Mouawad, Rima et al. (2015) Genome-Wide Analysis of Drosophila RBf2 Protein Highlights the Diversity of RB Family Targets and Possible Role in Regulation of Ribosome Biosynthesis. G3 (Bethesda) 5:1503-15
Sengupta, Satyaki; Lingnurkar, Raj; Carey, Timothy S et al. (2015) The Evolutionarily Conserved C-terminal Domains in the Mammalian Retinoblastoma Tumor Suppressor Family Serve as Dual Regulators of Protein Stability and Transcriptional Potency. J Biol Chem 290:14462-75
Sengupta, Satyaki; Henry, R William (2015) Regulation of the retinoblastoma-E2F pathway by the ubiquitin-proteasome system. Biochim Biophys Acta 1849:1289-97
Elenbaas, Jared S; Mouawad, Rima; Henry, R William et al. (2015) Role of Drosophila retinoblastoma protein instability element in cell growth and proliferation. Cell Cycle 14:589-97
Zhang, Liang; Wei, Yiliang; Pushel, Irina et al. (2014) Integrated stability and activity control of the Drosophila Rbf1 retinoblastoma protein. J Biol Chem 289:24863-73
Gjidoda, Alison; Henry, R William (2013) RNA polymerase III repression by the retinoblastoma tumor suppressor protein. Biochim Biophys Acta 1829:385-92
Raj, Nitin; Zhang, Liang; Wei, Yiliang et al. (2012) Rbf1 degron dysfunction enhances cellular DNA replication. Cell Cycle 11:3731-8
Raj, Nitin; Zhang, Liang; Wei, Yiliang et al. (2012) Ubiquitination of retinoblastoma family protein 1 potentiates gene-specific repression function. J Biol Chem 287:41835-43
Acharya, Pankaj; Negre, Nicolas; Johnston, John et al. (2012) Evidence for autoregulation and cell signaling pathway regulation from genome-wide binding of the Drosophila retinoblastoma protein. G3 (Bethesda) 2:1459-72

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