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首页> 美国卫生研究院文献>Cell Regulation >SCFCdc4-mediated Degradation of the Hac1p Transcription Factor Regulates the Unfolded Protein Response in Saccharomyces cerevisiae
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SCFCdc4-mediated Degradation of the Hac1p Transcription Factor Regulates the Unfolded Protein Response in Saccharomyces cerevisiae

机译：SCFCdc4介导的Hac1p转录因子的降解调节酿酒酵母中未折叠的蛋白质反应。

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摘要

The Saccharomyces cerevisiae basic leucine zipper transcription factor Hac1p is synthesized in response to the accumulation of unfolded polypeptides in the lumen of the endoplasmic reticulum (ER), and it is responsible for up-regulation of ∼5% of all yeast genes, including ER-resident chaperones and protein-folding catalysts. Hac1p is one of the most short-lived yeast proteins, having a half-life of ∼1.5 min. Here, we have shown that Hac1p harbors a functional PEST degron and that degradation of Hac1p by the proteasome involves the E2 ubiquitin-conjugating enzyme Ubc3/Cdc34p and the SCFCdc4 E3 complex. Consistent with the known nuclear localization of Cdc4p, rapid degradation of Hac1p requires the presence of a functional nuclear localization sequence, which we demonstrated to involve basic residues in the sequence 29RKRAKTK35. Two-hybrid analysis demonstrated that the PEST-dependent interaction of Hac1p with Cdc4p requires Ser146 and Ser149. Turnover of Hac1p may be dependent on transcription because it is inhibited in cell mutants lacking Srb10 kinase, a component of the SRB/mediator module of the RNA polymerase II holoenzyme. Stabilization of Hac1p by point mutation or deletion, or as the consequence of defects in components of the degradation pathway, results in increased unfolded protein response element-dependent transcription and improved cell viability under ER stress conditions.
机译：酿酒酵母碱性亮氨酸拉链转录因子Hac1p是根据内质网（ER）内腔中未折叠多肽的积累而合成的，它负责上调约5％的所有酵母基因，包括ER-常驻分子伴侣和蛋白质折叠催化剂。 Hac1p是寿命最短的酵母蛋白之一，半衰期约为1.5分钟。在这里，我们表明Hac1p带有功能性PEST degron，蛋白酶体降解Hac1p涉及E2泛素结合酶Ubc3 / Cdc34p和SCF Cdc4 E3复合物。与已知的Cdc4p核定位一致，Hac1p的快速降解需要功能性核定位序列的存在，我们证明了该序列涉及29RKRAKTK35序列中的基本残基。两杂交分析表明，Hac1p与Cdc4p的PEST依赖性相互作用需要Ser146和Ser149。 Hac1p的营业额可能取决于转录，因为它在缺少Srb10激酶的细胞突变体中受到抑制，而Srb10激酶是RNA聚合酶II全酶的SRB /介体模块的组成部分。 Hac1p通过点突变或缺失或降解途径组分缺陷的结果而稳定，导致内质网应激条件下未折叠的蛋白反应元件依赖性转录增加，细胞活力增强。

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