a Cato Research Ltd. , Durham , NC , USA.
b Institute of Metabolic Disorders, Genesis Biotechnology Group , Hamilton , NJ , USA.
Cell Cycle. 2017;16(22):2192-2203. doi: 10.1080/15384101.2017.1329065. Epub 2017 Sep 7.
Ceramides and sphingolipid intermediates are well-established regulators of the cell cycle. In the budding yeast Saccharomyces cerevisae, the complex sphingolipid backbone, ceramide, comprises a long chain sphingoid base, a polar head group, and a very long chain fatty acid (VLCFA). While ceramides and long chain bases have been extensively studied as to their roles in regulating cell cycle arrest under multiple conditions, the roles of VLCFAs are not well understood. Here, we used the yeast elo2 and elo3 mutants, which are unable to elongate fatty acids, as tools to explore if maintaining VLCFA elongation is necessary for cell cycle arrest in response to yeast mating. We found that both elo2 and elo3 cells had severely reduced mating efficiencies and were unable to form polarized shmoo projections that are necessary for cell-cell contact during mating. They also lacked functional MAP kinase signaling activity and were defective in initiating a cell cycle arrest in response to pheromone. Additional data suggests that mislocalization of the Ste5 scaffold in elo2 and elo3 mutants upon mating initiation may be responsible for the inability to initiate a cell cycle arrest. Moreover, the lack of proper Ste5 localization may be caused by the inability of mutant cells to mobilize PIP. We suggest that VLCFAs are required for Ste5 localization, which is a necessary event for initiating MAP kinase signaling and cell cycle arrest during yeast mating initiation.
神经酰胺和鞘脂中间产物是细胞周期的重要调节因子。在 budding yeast Saccharomyces cerevisiae 中,复杂的鞘脂骨干包括长链神经酰胺,极性头部基团和非常长链脂肪酸(VLCFA)。尽管已经广泛研究了神经酰胺和长链碱基在多种条件下调节细胞周期停滞的作用,但 VLCFA 的作用尚不清楚。在这里,我们使用无法延长脂肪酸的酵母 elo2 和 elo3 突变体作为工具,探讨维持 VLCFA 延长是否对于响应酵母交配而发生的细胞周期停滞是必需的。我们发现 elo2 和 elo3 细胞的交配效率均严重降低,并且无法形成极化的 shmoo 突起,这是交配过程中细胞间接触所必需的。它们还缺乏功能性 MAP 激酶信号转导活性,并且无法响应交配因子启动细胞周期停滞。其他数据表明,在交配起始时 elo2 和 elo3 突变体中 Ste5 支架的定位错误可能是无法启动细胞周期停滞的原因。此外,突变细胞无法动员 PIP 可能导致 Ste5 定位不当。我们认为 VLCFA 对于 Ste5 定位是必需的,这是在酵母交配起始时启动 MAP 激酶信号转导和细胞周期停滞的必要事件。