Analysis of the c-MYC-associated protein ZNF281

原型c-myc癌基因编码一种转录因子，这是诱导细胞增殖所必需的，也是足够的。C-myc蛋白的活性受多种相互作用蛋白的调节和调节。通过蛋白质组学方法，我们最近鉴定了锌指蛋白ZNF281/ZBP-99是一种c-myc相互作用蛋白。ZNF281直接与具有重要功能的MYC-Box II结构域结合。由于ZNF281对c-myc有较高的亲和力，我们认为这种相互作用对c-myc的功能非常重要。有趣的是，异位ZNF281抑制c-myc介导的基因激活并抑制细胞增殖。在这项提议中，我们打算确定c-MYC/ZNF281相互作用在哪个分子背景下是功能相关的。因此，ZNF281的上游信号转导将通过蛋白质组鉴定和ZNF281相关蛋白的功能分析来确定。此外，还将分析MYC/ZNF281相互作用在基因调控中的作用。因此，我们将在全基因组水平上比较ZNF281、c-MYC或两者联合激活后ZNF281和c-MYC的差异基因调控和启动子占位情况。这种比较将使识别受这两个因素共同调控的基因成为可能。随后，将对这些基因进行功能分析，以确定它们在介导ZNF281/c-myc复合体对细胞的影响中所起的作用。

The prototypic c-MYC oncogene encodes a transcription factor which is required and sufficient for the induction of cellular proliferation. The activity of the c-MYC protein is regulated and mediated by numerous interacting proteins. Via a proteomic approach we recently identified the zinc-finger protein ZNF281/ZBP-99 as a c-MYC interacting protein. ZNF281 directly binds to the functionally important MYC-Box II domain. Since ZNF281 has a comparatively high affinity to c-MYC, we expect that this interaction is of great importance for the function of c-MYC. Interestingly, ectopic ZNF281 represses c-MYC-mediated gene activation and inhibits cell proliferation. In this proposal we intend to determine in which molecular context the c-MYC/ZNF281 interaction is functionally relevant. Therefore, the upstream signaling of ZNF281 will be characterized by proteomic identification and functional analysis of ZNF281-associated proteins. In addition, the role of the MYC/ZNF281 interaction in gene regulation will be analyzed. Therefore, differential gene regulation and promoter occupation of ZNF281 and c-MYC after activation of ZNF281, c-MYC or a combination of both will be compared on a genome-wide scale. This comparison will allow the identification of genes co-regulated by both factors. Subsequently, functional analysis of such genes will be performed to determine their role in mediating the effect of a ZNF281/c-MYC complex on cells.