Hapke Robert, Venton Lindsay, Rose Kristie Lindsay, Sheng Quanhu, Reddy Anupama, Prather Rebecca, Jones Angela, Rathmell W Kimryn, Haake Scott M
Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Department of Genetic Medicine, Vanderbilt University Medical Center, Nashville, TN, USA.
Kidney Cancer J. 2022;6(3):179-193. doi: 10.3233/kca-220009. Epub 2022 Nov 18.
SET domain-containing protein 2 () is commonly mutated in renal cell carcinoma. SETD2 methylates histone H3 as well as a growing list of non-histone proteins.
Initially, we sought to explore SETD2-dependent changes in lysine methylation of proteins in proximal renal tubule cells. Subsequently, we focused on changes in lysine methylation of the translation elongation factor eEF1A1.
To accomplish these objectives, we initially performed a systems-wide analysis of protein lysine-methylation and expression in wild type (WT) and -knock out (KO) kidney cells and later focused our studies on eEF1A1 as well as the expression of lysine methyltransferases that regulate its lysine methylation.
We observed decreased lysine methylation of the translation elongation factor eEF1A1. and are known to methylate eEF1A1, and we show here that their expression is dependent on SET-domain function of Globally, we observe differential expression of hundreds of proteins in WT versus -KO cells, including increased expression of many involved in protein translation. Finally, we observe decreased progression free survival and loss of EEF1AKMT2 gene expression in -mutated tumors predicted to have loss of function of the SET domain.
Overall, these data suggest that -mutated ccRCC, via loss of enzymatic function of the SET domain, displays dysregulation of protein translation as a potentially important component of the transformed phenotype.
含SET结构域蛋白2（SETD2）在肾细胞癌中常见突变。SETD2可使组蛋白H3以及越来越多的非组蛋白发生甲基化。
最初，我们试图探究近端肾小管细胞中SETD2依赖性蛋白质赖氨酸甲基化的变化。随后，我们聚焦于翻译延伸因子eEF1A1赖氨酸甲基化的变化。
为实现这些目标，我们首先对野生型（WT）和SETD2基因敲除（KO）肾细胞中的蛋白质赖氨酸甲基化和表达进行全系统分析，之后将研究重点放在eEF1A1以及调节其赖氨酸甲基化的赖氨酸甲基转移酶的表达上。
我们观察到翻译延伸因子eEF1A1的赖氨酸甲基化减少。已知SETD2和KMT2C可使eEF1A1甲基化，我们在此表明它们的表达依赖于SETD2的SET结构域功能。总体而言，我们观察到WT细胞与SETD2基因敲除细胞中数百种蛋白质的差异表达，包括许多参与蛋白质翻译的蛋白质表达增加。最后，我们观察到在预测SET结构域功能丧失的SETD2突变肿瘤中，无进展生存期缩短且EEF1AKMT2基因表达缺失。
总体而言，这些数据表明，SETD2突变的透明细胞肾细胞癌通过SET结构域酶功能丧失，表现出蛋白质翻译失调，这可能是转化表型的一个重要组成部分。
