Department of Biochemistry, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan.
Proc Natl Acad Sci U S A. 2014 Jan 14;111(2):699-704. doi: 10.1073/pnas.1316433111. Epub 2013 Dec 27.
Currently, there is no method to distinguish between the roles of a subunit in one multisubunit protein complex from its roles in other complexes in vivo. This is because a mutation in a common subunit will affect all complexes containing that subunit. Here, we describe a unique method to discriminate between the functions of a common subunit in different multisubunit protein complexes. In this method, a common subunit in a multisubunit protein complex is genetically fused to a subunit that is specific to that complex and point mutated. The resulting phenotype(s) identify the specific function(s) of the subunit in that complex only. Histone H2B is a common subunit in nucleosomes containing H2A/H2B or Htz1/H2B dimers. The H2B was fused to H2A or Htz1 and point mutated. This strategy revealed that H2B has common and distinct functions in different nucleosomes. This method could be used to study common subunits in other multisubunit protein complexes.
目前,尚无方法可以区分一个多亚基蛋白复合物中一个亚基的作用与其在体内其他复合物中的作用。这是因为常见亚基中的突变会影响所有包含该亚基的复合物。在这里,我们描述了一种独特的方法,可以区分不同多亚基蛋白复合物中常见亚基的功能。在该方法中,将多亚基蛋白复合物中的常见亚基基因融合到该复合物特有的且定点突变的亚基上。由此产生的表型仅确定该亚基在该复合物中的特定功能。组蛋白 H2B 是含有 H2A/H2B 或 Htz1/H2B 二聚体的核小体中的常见亚基。将 H2B 融合到 H2A 或 Htz1 并进行定点突变。该策略揭示了 H2B 在不同核小体中有共同和独特的功能。该方法可用于研究其他多亚基蛋白复合物中的常见亚基。