Department of Cell Biology, Faculty of Science, Charles University, Prague, Czech Republic.
Department of Computing and Control Engineering, University of Chemistry and Technology, Prague, Czech Republic.
FEMS Yeast Res. 2018 Sep 1;18(6). doi: 10.1093/femsyr/foy064.
Fission yeast 'cut' mutants show defects in temporal coordination of nuclear division with cytokinesis, resulting in aberrant mitosis and lethality. Among other causes, the 'cut' phenotype can be triggered by genetic or chemical perturbation of lipid metabolism, supposedly resulting in shortage of membrane phospholipids and insufficient nuclear envelope expansion during anaphase. Interestingly, penetrance of the 'cut' phenotype in mutants of the transcription factor cbf11 and acetyl-coenzyme A carboxylase cut6, both related to lipid metabolism, is highly dependent on growth media, although the specific nutrient(s) affecting 'cut' occurrence is not known. In this study, we set out to identify the growth media component(s) responsible for 'cut' phenotype suppression in Δcbf11 and cut6-621 cells. We show that mitotic defects occur rapidly in Δcbf11 cells upon shift from the minimal EMM medium ('cut' suppressing) to the complex YES medium ('cut' promoting). By growing cells in YES medium supplemented with individual EMM components, we identified ammonium chloride, an efficiently utilized nitrogen source, as a specific and potent suppressor of the 'cut' phenotype in both Δcbf11 and cut6-621. Furthermore, we found that ammonium chloride boosts lipid droplet formation in wild-type cells. Our findings suggest a possible involvement of nutrient-responsive signaling in 'cut' suppression.
裂殖酵母“切割”突变体显示出核分裂与胞质分裂的时间协调缺陷,导致异常有丝分裂和致死。除其他原因外,“切割”表型可由脂质代谢的遗传或化学扰动引发,推测这导致了后期膜磷脂短缺和核膜扩张不足。有趣的是,转录因子 cbf11 和乙酰辅酶 A 羧化酶 cut6 的突变体的“切割”表型的外显率高度依赖于生长培养基,尽管影响“切割”发生的具体营养物质尚不清楚。在这项研究中,我们着手确定负责Δcbf11 和 cut6-621 细胞中“切割”表型抑制的生长培养基成分。我们表明,Δcbf11 细胞从最小的 EMM 培养基(“切割”抑制)转移到复杂的 YES 培养基(“切割”促进)时,有丝分裂缺陷会迅速发生。通过在 YES 培养基中补充单个 EMM 成分来培养细胞,我们鉴定出氯化铵,一种有效利用的氮源,是Δcbf11 和 cut6-621 中“切割”表型的特异性和有效抑制剂。此外,我们发现氯化铵可促进野生型细胞中脂滴的形成。我们的研究结果表明,营养物质响应信号可能参与了“切割”抑制。