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

聚(ADP-核糖)聚合酶(PARP)是一小类利用NAD+对底物蛋白质进行ADPriBosylate的酶。PARP酶控制基因转录、DNA修复、遗传稳定性和细胞增殖的不同方面。我们通过与原癌蛋白c-Myc的相互作用鉴定出PARP-10是该家族的一个新成员，原癌蛋白c-Myc是一种转录调节因子，控制着广泛的细胞活动。PARP-10干扰原代细胞Myc依赖性转化。这取决于它从细胞核输出的能力，而不是它的PARP活性。为了更详细地研究PARP-10的功能并最终了解其对Myc功能的贡献，我们打算使用TAP-标签系统结合质谱学来鉴定和表征PARP-10相关蛋白。已确定的相关蛋白将被分析它们在PARP-10和Myc的调节和功能中的作用。由于PARP-10的核/细胞质穿梭在功能上很重要，我们将讨论这一过程的机制和调控。为此，我们将使用不同的方法，包括光漂白或翻转中的荧光损失。通过这些研究，我们希望确定PARP-10的基本特征，并有助于更好地理解Myc。

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.