Conserved Mode of Interaction between Yeast Bro1 Family V Domains and YP(X)nL Motif-Containing Target Proteins
Conserved Mode of Interaction between Yeast Bro1 Family V Domains and YP(X)nL Motif-Containing Target Proteins
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DOI：
10.1128/ec.00091-15
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发表时间：
2015-10-01
期刊：
影响因子：
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通讯作者：
Tanaka, Keiji
中科院分区：
文献类型：
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作者：
Kimura, Yoko;Tanigawa, Mirai;Tanaka, Keiji
酵母菌Bro1和Rim20属于一个蛋白质家族，具有Bro1和V结构域的共同结构。Alix和His结构域蛋白酪氨酸磷酸酶(HD-PTP)是哺乳动物Bro1家族蛋白，通过其V结构域与靶蛋白中的Yp(X)NL(n=1~3)基序结合。在Alix中，位于V结构域疏水槽中的Phe残基是与YP(X)NL基序结合的关键。虽然哺乳动物和酵母V结构域之间的总序列不是高度保守的，但我们表明酵母Bro1 V结构域中保守的Phe残基对于与其含有YP(X)NL的靶蛋白Rfu1结合是重要的。此外，我们还发现Rim20通过Rim20的V结构域与Rim101的YPIKL基序相互作用而与其靶蛋白Rim101结合。Rim20V结构域的关键Phe残基或Rim101的YPIKL基序的突变影响了Rim20介导的Rim101的加工。这些结果表明，V结构域与YP(X)NL基序蛋白之间的相互作用从酵母到哺乳动物细胞都是保守的。此外，每个V结构域对其目标蛋白的特异性表明，未知元件决定了结合特异性。
Yeast Bro1 and Rim20 belong to a family of proteins which possess a common architecture of Bro1 and V domains. Alix and His domain protein tyrosine phosphatase (HD-PTP), mammalian Bro1 family proteins, bind YP(X)nL (n = 1 to 3) motifs in their target proteins through their V domains. In Alix, the Phe residue, which is located in the hydrophobic groove of the V domain, is critical for binding to the YP(X)nL motif. Although the overall sequences are not highly conserved between mammalian and yeast V domains, we show that the conserved Phe residue in the yeast Bro1 V domain is important for binding to its YP(X)nL-containing target protein, Rfu1. Furthermore, we show that Rim20 binds to its target protein Rim101 through the interaction between the V domain of Rim20 and the YPIKL motif of Rim101. The mutation of either the critical Phe residue in the Rim20 V domain or the YPIKL motif of Rim101 affected the Rim20-mediated processing of Rim101. These results suggest that the interactions between V domains and YP(X)nL motif-containing proteins are conserved from yeast to mammalian cells. Moreover, the specificities of each V domain to their target protein suggest that unidentified elements determine the binding specificity.