Ras oncogene induced protein SUMOylation changes

研究背景通过RNAi合成致死筛选，我们已经鉴定了Ras突变癌细胞生存所需的蛋白SUMO化途径。在本项目中，我们旨在解决以下问题：1）抑制Ras突变的SUMO化途径是合成致死的机制; 2）哪些细胞蛋白在Ras突变细胞中被差异SUMO化;重要材料和方法1）靶向SUMO E1和E2连接酶以及SUMO途径蛋白的shRNAs; 2）表达SUMO蛋白和连接酶的稳定细胞系; 3）用于鉴定细胞裂解物中SUMO化蛋白质的质谱方案。FY 2012 ACCOMPLISHMENT 2011财年继续我们的努力，我们已经鉴定了许多在Ras WT和突变细胞之间差异SUMO化的候选蛋白质。我们目前正在研究这些SUMO化蛋白在KRAS突变细胞中的功能。我们还在研究SUMO化如何影响这些蛋白质的活性。

BACKGROUNDThrough an RNAi synthetic lethal screen we have identified the protein SUMOylation pathway to be required for the viability of Ras mutant cancer cells.PURPOSEIn this project we aim to address the following questions: 1) the mechanism by which the inhibition of SUMOylation pathway is synthetically lethal with Ras mutation; 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 METHODS1) 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 protocol for identifying SUMOylated proteins in cell lysates.FY2012 ACCOMPLISHMENTContinuing our effort from FY2011, we have identified a number of candidate proteins that are differentially SUMOylated between Ras WT and mutant cells. We are currently studying the functions of these SUMOylated proteins in KRAS mutant cells. We are also investigating how SUMOylation affects the activity of these proteins.