Dynamic localization of protein phosphatase type 1 in the mitotic cell cycle of Saccharomyces cerevisiae.
Dynamic localization of protein phosphatase type 1 in the mitotic cell cycle of Saccharomyces cerevisiae.
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DOI：
10.1083/jcb.149.1.125
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发表时间：
2000-04-03
期刊：
影响因子：
--
通讯作者：
Tatchell K
中科院分区：
文献类型：
--
作者：
Bloecher A;Tatchell K
蛋白磷酸酶I（PP 1），由酿酒酵母中的单一必需基因GLC 7编码，在多种细胞过程中发挥作用。为了鉴定这些过程发生的体内亚细胞位置，我们使用了功能性绿色荧光蛋白（GFP）-Glc 7 p融合蛋白。延时荧光显微镜显示GFP-Glc 7 p主要定位于整个有丝分裂细胞周期的细胞核中，在核仁中浓度最高。GFP-Glc 7 p也在芽颈处的环中观察到，这依赖于功能性隔蛋白。Glc 7 -129突变体显示随机出芽模式，支持Glc 7 p在芽位选择中的作用。在α-因子处理的细胞中，GFP-Glc 7 p位于交配投射的基部，再次以隔蛋白依赖性方式。在分裂后期开始时，GFP-Glc 7 p在纺锤体极体处积累并保持在那里直到胞质分裂。后期后，GFP-Glc 7 p集中在与肌动球蛋白环共定位的环中。GFP-Glc 7 -129融合体在芽颈和SPB的定位上有缺陷。总之，这些结果确定了Glc 7 p功能的位点，并表明Glc 7 p活性通过其位置的动态变化来调节。
Protein phosphatase type I (PP1), encoded by the single essential gene GLC7 in Saccharomyces cerevisiae, functions in diverse cellular processes. To identify in vivo subcellular location(s) where these processes take place, we used a functional green fluorescent protein (GFP)–Glc7p fusion protein. Time-lapse fluorescence microscopy revealed GFP–Glc7p localizes predominantly in the nucleus throughout the mitotic cell cycle, with the highest concentrations in the nucleolus. GFP–Glc7p was also observed in a ring at the bud neck, which was dependent upon functional septins. Supporting a role for Glc7p in bud site selection, a glc7-129 mutant displayed a random budding pattern. In α-factor treated cells, GFP–Glc7p was located at the base of mating projections, again in a septin-dependent manner. At the start of anaphase, GFP–Glc7p accumulated at the spindle pole bodies and remained there until cytokinesis. After anaphase, GFP–Glc7p became concentrated in a ring that colocalized with the actomyosin ring. A GFP–Glc7-129 fusion was defective in localizing to the bud neck and SPBs. Together, these results identify sites of Glc7p function and suggest Glc7p activity is regulated through dynamic changes in its location.
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