Division of Gene Structure and Function, Research Center for Genomic Medicine, Saitama Medical University, 1397-1 Yamane, Hidaka-shi, Saitama-ken 350-1241, Japan.
Biochem Biophys Res Commun. 2011 Jan 28;404(4):991-6. doi: 10.1016/j.bbrc.2010.12.097. Epub 2010 Dec 25.
TLS (Translocated in LipoSarcoma), also termed FUS, is a multifunctional protein implicated in diverse cellular events such as maintaining genome integrity and regulating gene expression. We have focused on the role of TLS as a coregulator in transcriptional regulation. In the process of investigating TLS-binding proteins, we found that PRMT1 (protein arginine methyltransferase 1) was in complex with TLS. We analyzed the methylation status of endogenous TLS and demonstrated that TLS was arginine-methylated by PRMT1. Using mass spectrometry, we identified that four arginine residues within TLS (R216, R218, R242 and R394) were consistently dimethylated. We performed luciferase reporter assays to assess the functional consequence of TLS arginine methylation in transcriptional regulation and, interestingly, observed that TLS and PRMT1 synergistically coactivated transcription at the survivin promoter. Further analysis using a catalytic-dead PRMT1 or methylation inhibitor both showed that the synergistic transcriptional activation was mediated by TLS arginine-methylation. These results revealed a cooperative role of TLS and PRMT1 in transcriptional regulation.
TLS（易位性脂肪肉瘤），也称为 FUS，是一种多功能蛋白，参与多种细胞事件，如维持基因组完整性和调节基因表达。我们专注于 TLS 作为转录调节核心调节剂的作用。在研究 TLS 结合蛋白的过程中，我们发现 PRMT1（蛋白质精氨酸甲基转移酶 1）与 TLS 形成复合物。我们分析了内源性 TLS 的甲基化状态，并证实 PRMT1 将 TLS 精氨酸甲基化。通过质谱分析，我们鉴定出 TLS 内的四个精氨酸残基（R216、R218、R242 和 R394）被一致地二甲基化。我们进行了荧光素酶报告基因检测，以评估 TLS 精氨酸甲基化在转录调节中的功能后果，有趣的是，观察到 TLS 和 PRMT1 协同激活存活素启动子的转录。进一步使用催化失活的 PRMT1 或甲基化抑制剂进行分析表明，协同转录激活是由 TLS 精氨酸甲基化介导的。这些结果揭示了 TLS 和 PRMT1 在转录调节中的协同作用。
