Vasicova Pavla, Rinnerthaler Mark, Haskova Danusa, Novakova Lenka, Malcova Ivana, Breitenbach Michael, Hasek Jiri
Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.
Department of Cell Biology, Division of Genetics, University of Salzburg, Salzburg, Austria.
Microb Cell. 2016 Apr 12;3(5):206-214. doi: 10.15698/mic2016.05.499.
Actin filaments form cortical patches and emanating cables in fermenting cells of . This pattern has been shown to be depolarized in glucose-depleted cells after formaldehyde fixation and staining with rhodamine-tagged phalloidin. Loss of actin cables in mother cells was remarkable. Here we extend our knowledge on actin in live glucose-depleted cells co-expressing the marker of actin patches (Abp1-RFP) with the marker of actin cables (Abp140-GFP). Glucose depletion resulted in appearance of actin patches also in mother cells. However, even after 80 min of glucose deprivation these cells showed a clear network of actin cables labeled with Abp140-GFP in contrast to previously published data. In live cells with a mitochondrial dysfunction (rho cells), glucose depletion resulted in almost immediate appearance of Abp140-GFP foci partially overlapping with Abp1-RFP patches in mother cells. Residual actin cables were clustered in patch-associated bundles. A similar overlapping "patchy" pattern of both actin markers was observed upon treatment of glucose-deprived rho cells with FCCP (the inhibitor of oxidative phosphorylation) and upon treatment with formaldehyde. While the formaldehyde-targeted process stays unknown, our results indicate that published data on yeast actin cytoskeleton obtained from glucose-depleted cells after fixation should be considered with caution.
肌动蛋白丝在……的发酵细胞中形成皮质斑和延伸的束状结构。在甲醛固定并用罗丹明标记的鬼笔环肽染色后，已证明这种模式在葡萄糖耗尽的细胞中会去极化。母细胞中肌动蛋白束状结构的丧失很明显。在这里，我们扩展了对共表达肌动蛋白斑标记物（Abp1-RFP）和肌动蛋白束状结构标记物（Abp140-GFP）的活的葡萄糖耗尽细胞中肌动蛋白的认识。葡萄糖耗尽也导致母细胞中出现肌动蛋白斑。然而，与先前发表的数据相反，即使在葡萄糖剥夺80分钟后，这些细胞仍显示出由Abp140-GFP标记的清晰的肌动蛋白束状网络。在具有线粒体功能障碍的活细胞（rho细胞）中，葡萄糖耗尽导致母细胞中几乎立即出现与Abp1-RFP斑部分重叠的Abp140-GFP焦点。残余的肌动蛋白束状结构聚集在与斑相关的束中。在用FCCP（氧化磷酸化抑制剂）处理葡萄糖剥夺的rho细胞以及用甲醛处理后，观察到两种肌动蛋白标记物都有类似的重叠“斑状”模式。虽然甲醛靶向的过程尚不清楚，但我们的结果表明，从固定后的葡萄糖耗尽细胞中获得的关于酵母肌动蛋白细胞骨架的已发表数据应谨慎考虑。
