Role of the protein SUMOylation pathway in Ras-driven oncogenesis

背景。通过RNAi合成致死筛选，我们已经确定了Ras突变癌细胞的生存和转化所需的蛋白质SUMOylation途径。目的。在这个项目中，我们的目标是解决以下问题：1)SUMO连接酶的抑制与KRAS癌基因合成致死的机制；2)哪些细胞蛋白在Ras突变细胞中被不同的SUMOylated；3)在Ras突变肿瘤中，这些蛋白SUMOylation状态的改变如何影响其功能。重要的材料和方法。1)靶向SUMO E1和E2连接酶及SUMO通路蛋白的shrna；2)表达SUMO蛋白和连接酶的稳定细胞系；3)鉴定细胞裂解物中SUMOylated蛋白的质谱方法。FY2015成就。我们发现KRAS突变的结直肠癌细胞表现出蛋白质SUMO化模式的改变，Ras驱动的转化需要SUMO连接酶活性。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.