The essential yeast RNA binding protein Np13p is methylated.
The essential yeast RNA binding protein Np13p is methylated.
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DOI：
10.1073/pnas.93.24.13641
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发表时间：
1996-11
影响因子：
11.1
通讯作者：
Christian W. Siebel;Christine Guthrie
中科院分区：
文献类型：
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作者：
Christian W. Siebel;Christine Guthrie
精氨酸甲基化是许多RNA结合蛋白中普遍存在的修饰，但对其功能后果知之甚少。使用单克隆抗体，1 E4，我们已经表明，酵母NPL 3基因产物Np 13 p，一个重要的RNA结合蛋白与重复RGG基序，是在体内甲基化的精氨酸。1 E4表位可以通过将重组Np 13 p与部分纯化的牛精氨酸甲基转移酶孵育来产生，阻断该反应。Np 13 p甲基化需要S-腺苷-L-甲硫氨酸，也发生在酵母提取物中。缺乏RGG结构域的Np 13 p缺失突变体不是甲基化的底物，这表明甲基化位点位于RGG基序内。精氨酸甲基化在遗传上易处理的生物体中的发现为理解这种修饰的功能提供了一个强有力的入口，特别是考虑到Np 13 p在RNA加工和转运中的许多作用。最近发现的磷酸化丝氨酸残基内的RGG域提出了一种假说，其中的分子开关由甲基化和磷酸化调节的Np 13 p RGG域的生化特性。
Arginine methylation is a prevalent modification found in many RNA binding proteins, yet little is known about its functional consequences. Using a monoclonal antibody, 1E4, we have shown that the yeast NPL3 gene product Np13p, an essential RNA binding protein with repeated RGG motifs, is arginine-methylated in vivo. The 1E4 epitope can be generated by incubating recombinant Np13p with partially purified bovine arginine methyltransferase block this reaction. Np13p methylation requires S-adenosyl-L-methionine and also occurs in yeast extracts. An Np13p deletion mutant lacking the RGG domain is not a substrate for methylation, suggesting that the methylation sites lie within the RGG motifs. The discovery of arginine methylation in a genetically tractable organism provides a powerful entrée to understanding the function of this modification, particularly in view of the many roles postulated for Np13p in RNA processing and transport. The recent discovery of phosphorylated serine residues within the RGG domain suggests a hypothesis in which a molecular switch governed by methylation and phosphorylation regulates the biochemical properties of the Np13p RGG domain.