Role of the protein SUMOylation pathway in Ras-driven oncogenesis

背景资料。通过RNAi合成致死筛选，我们已经确定了RAS突变癌细胞存活和转化所需的蛋白质SUMO化途径。目的。在这个项目中，我们旨在解决以下问题：1)抑制相扑连接酶与KRAS癌基因合成致死的机制；2)在RAS突变细胞中，哪些细胞蛋白发生差异SUMO化；以及3)在RAS突变肿瘤的背景下，这些蛋白的SUMO化状态的变化如何影响它们的功能。重要的材料和方法。1)针对相扑E1和E2连接酶和相扑途径蛋白的shRNA；2)稳定表达相扑蛋白和连接酶的细胞系；3)鉴定细胞裂解产物中SUMO化蛋白的质谱学方法。2015财年成就。我们发现，KRAS突变的结直肠癌细胞表现出蛋白SUMO化模式的改变，而RAS驱动的转化需要相扑连接酶的活性。这项研究发表于2015年。

BACKGROUND. Through a RNAi synthetic lethal screen we have identified the protein SUMOylation pathway to be required for the viability and transformation of Ras mutant cancer cells. PURPOSE. In this project we aim to address the following questions: 1) the mechanism by which inhibition of SUMO ligases is synthetically lethal with the KRAS oncogene; 2) which cellular proteins are differentially SUMOylated in Ras mutant cells; and 3) How the changes in SUMOylation status of these proteins affect their function in the context of Ras mutant tumors. SIGNIFICANT MATERIALS AND METHODS. 1) shRNAs that target the SUMO E1 and E2 ligases and SUMO pathway proteins; 2) Stable cell lines expressing SUMO proteins and ligases; 3) mass-spectrometry methodology for identifying SUMOylated proteins in cell lysates. FY2015 ACCOMPLISHMENT. We discovered that KRAS mutant colorectal cancer cells exhibit altered patterns of protein SUMOylation and Ras driven transformation requires SUMO ligase activity. This study was published in 2015.