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首页> 外文期刊>Molecular and Cellular Biology >The EH-domain-containing protein Pan1 is required for normal organization of the actin cytoskeleton in Saccharomyces cerevisiae.
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The EH-domain-containing protein Pan1 is required for normal organization of the actin cytoskeleton in Saccharomyces cerevisiae.

机译：酿酒酵母中，肌动蛋白细胞骨架的正常组织需要含有EH结构域的蛋白Pan1。

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Normal cell growth and division in the yeast Saccharomyces cerevisiae involve dramatic and frequent changes in the organization of the actin cytoskeleton. Previous studies have suggested that the reorganization of the actin cytoskeleton in accordance with cell cycle progression is controlled, directly or indirectly, by the cyclin-dependent kinase Cdc28. Here we report that by isolating rapid-death mutants in the background of the Start-deficient cdc28-4 mutation, the essential yeast gene PAN1, previously thought to encode the yeast poly(A) nuclease, is identified as a new factor required for normal organization of the actin cytoskeleton. We show that at restrictive temperature, the pan1 mutant exhibited abnormal bud growth, failed to maintain a proper distribution of the actin cytoskeleton, was unable to reorganize actin the cytoskeleton during cell cycle, and was defective in cytokinesis. The mutant also displayed a random pattern of budding even at permissive temperature. Ectopic expression of PAN1 by the GAL promoter caused abnormal distribution of the actin cytoskeleton when a single-copy vector was used. Immunofluorescence staining revealed that the Pan1 protein colocalized with the cortical actin patches, suggesting that it may be a filamentous actin-binding protein. The Pan1 protein contains an EF-hand calcium-binding domain, a putative Src homology 3 (SH3)-binding domain, a region similar to the actin cytoskeleton assembly control protein Sla1, and two repeats of a newly identified protein motif known as the EH domain. These findings suggest that Pan1, recently recognized as not responsible for the poly(A) nuclease activity (A. B. Sachs and J. A. Deardorff, erratum, Cell 83:1059, 1995; R. Boeck, S. Tarun, Jr., M. Rieger, J. A. Deardorff, S. Muller-Auer, and A. B. Sachs, J. Biol. Chem. 271:432-438, 1996), plays an important role in the organization of the actin cytoskeleton in S. cerevisiae.
机译：酿酒酵母中正常细胞的生长和分裂涉及肌动蛋白细胞骨架组织的剧烈而频繁的变化。先前的研究表明，根据细胞周期进展，肌动蛋白细胞骨架的重组是直接或间接地由细胞周期蛋白依赖性激酶Cdc28控制的。在这里，我们报告说，通过在缺乏启动子的cdc28-4突变的背景中分离快速死亡突变体，先前被认为编码酵母多核糖核酸酶的必需酵母基因PAN1被确定为正常人必需的新因子。肌动蛋白细胞骨架的组织。我们表明，在限制性温度下，pan1突变体表现出异常的芽生长，未能维持肌动蛋白细胞骨架的适当分布，在细胞周期中无法重组肌动蛋白细胞骨架，并且在胞质分裂中存在缺陷。该突变体即使在允许的温度下也显示出随机的出芽模式。当使用单拷贝载体时，GAL启动子异位表达PAN1会导致肌动蛋白细胞骨架的异常分布。免疫荧光染色显示Pan1蛋白与皮质肌动蛋白斑块共定位，表明它可能是丝状肌动蛋白结合蛋白。 Pan1蛋白包含一个EF手钙结合结构域，一个假定的Src同源性3（SH3）结合结构域，一个类似于肌动蛋白细胞骨架装配控制蛋白Sla1的区域，以及两个新发现的称为EH的蛋白基序的重复序列域。这些发现表明Pan1最近被认为与poly（A）核酸酶活性无关（AB Sachs和JA Deardorff，勘误表，Cell 83：1059，1995; R.Boeck，S.Tarun，Jr.，M.Rieger， JA Deardorff，S.Muller-Auer和AB Sachs，J.Biol.Chem.271：432-438，1996）在酿酒酵母中肌动蛋白细胞骨架的组织中起重要作用。

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