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首页> 外文期刊>Molecular and Cellular Biology >Genome-Wide Analysis Identifies MYND-Domain Protein Mub1 as an Essential Factor for Rpn4 Ubiquitylation
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Genome-Wide Analysis Identifies MYND-Domain Protein Mub1 as an Essential Factor for Rpn4 Ubiquitylation

机译：全基因组分析确定MYND域蛋白Mub1为Rpn4泛素化的必要因素。

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The proteasome homeostasis in Saccharomyces cerevisiae is regulated by a negative feedback circuit in which the Rpn4 transcription factor upregulates the proteasome genes and is rapidly degraded by the proteasome. Previous work has identified Ubr2 and Rad6 as the cognate E3 and E2 enzymes for Rpn4 ubiquitylation. However, our recent attempts to ubiquitylate Rpn4 using purified Ubr2 and Rad6 proteins in a reconstitution system have been unsuccessful, suggesting that an additional factor is required for Rpn4 ubiquitylation. Here, we screened the entire collection of the single-gene-deletion yeast mutants generated by the Saccharomyces Genome Deletion Project and identified the mub1Δ mutant defective in ubiquitin-dependent degradation of Rpn4. An in vitro reconstitution ubiquitylation assay confirms that Mub1 is the missing factor for Rpn4 ubiquitylation. We further show that Mub1 directly interacts with Ubr2 and Rpn4. The MYND domain of Mub1 may play an important role in Rpn4 ubiquitylation. Interestingly, Mub1 itself is a short-lived protein and its degradation is dependent on the Ubr2/Rad6 ubiquitin ligase. Together, these data suggest that Mub1 and Ubr2 cooperate to transfer ubiquitin to Rpn4 from Rad6 and that Mub1 may switch from a partner to a substrate of the Ubr2/Rad6 ubiquitin ligase.
机译：酿酒酵母（Saccharomyces cerevisiae）中的蛋白酶体稳态由负反馈电路调节，其中Rpn4转录因子上调了蛋白酶体基因，并被蛋白酶体迅速降解。先前的工作已将Ubr2和Rad6识别为Rpn4泛素化的同源E3和E2酶。但是，我们最近在重组系统中使用纯化的Ubr2和Rad6蛋白泛素化Rpn4的尝试仍未成功，这表明Rpn4泛素化需要其他因子。在这里，我们筛选了由 Saccharomyces 基因组缺失计划产生的单基因缺失酵母突变体的整个集合，并鉴定了在泛素依赖性降解中的 mub1 Δ突变体。 Rpn4。体外重组泛素化测定证实Mub1是Rpn4泛素化的缺失因子。我们进一步证明Mub1与Ubr2和Rpn4直接交互。 Mub1的MYND域可能在Rpn4泛素化中起重要作用。有趣的是，Mub1本身是一种寿命短的蛋白质，其降解取决于Ubr2 / Rad6泛素连接酶。在一起，这些数据表明Mub1和Ubr2协同将泛素从Rad6转移到Rpn4，并且Mub1可能从伴侣转变为Ubr2 / Rad6泛素连接酶的底物。

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