Characterization and function of the poly(ADP-ribose) polymerase PARP-10 in the control of cell behavior

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Poly(ADP-ribose) polymerases (PARP) are a small family of enzymes that use NAD+ to ADPribosylate substrate proteins. PARP enzymes control different aspects of gene transcription, DNA repair, genetic stability and cell proliferation. We have identified with PARP-10 a novel member of the family through its interaction with the proto-oncoprotein c-Myc, a transcriptional regulator that controls a broad spectrum of cellular activities. PARP-10 interferes with Myc-dependent transformation of primary cells. This is dependent on its ability to be exported from the nucleus but not on its PARP activity. To address the function of PARP-10 in more detail and ultimately to understand its contribution to Myc function, we intend to identify and characterize PARP-10-associated proteins using the TAP-tagging system in combination with mass spectrometry. Identified associated proteins will be analyzed for their role in the regulation and function of PARP-10 and Myc. Since nuclear/cytoplasmic shuttling of PARP-10 is functionally important we will address the mechanism and the regulation of this process. For this we will use different approaches, including Fluorescence Loss In Photobleaching or FLIP. With these studies we expect to define basic characteristics of PARP-10 and to contribute to the better understanding of Myc.