Luo Jun, Matsuo Yasuhiro, Gulis Galina, Hinz Haylee, Patton-Vogt Jana, Marcus Stevan
Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL 35487, USA.
Eukaryot Cell. 2009 May;8(5):790-9. doi: 10.1128/EC.00029-09. Epub 2009 Mar 13.
To investigate the contributions of phosphatidylethanolamine to the growth and morphogenesis of the fission yeast Schizosaccharomyces pombe, we have characterized three predicted genes in this organism, designated psd1, psd2, and psd3, encoding phosphatidylserine decarboxylases, which catalyze the conversion of phosphatidylserine to phosphatidylethanolamine in both eukaryotic and prokaryotic organisms. S. pombe mutants carrying deletions in any one or two psd genes are viable in complex rich medium and synthetic defined minimal medium. However, mutants carrying deletions in all three psd genes (psd1-3Delta mutants) grow slowly in rich medium and are inviable in minimal medium, indicating that the psd1 to psd3 gene products share overlapping essential cellular functions. Supplementation of growth media with ethanolamine, which can be converted to phosphatidylethanolamine by the Kennedy pathway, restores growth to psd1-3Delta cells in minimal medium, indicating that phosphatidylethanolamine is essential for S. pombe cell growth. psd1-3Delta cells produce lower levels of phosphatidylethanolamine than wild-type cells, even in medium supplemented with ethanolamine, indicating that the Kennedy pathway can only partially compensate for the loss of phosphatidylserine decarboxylase activity in S. pombe. psd1-3Delta cells appear morphologically indistinguishable from wild-type S. pombe cells in medium supplemented with ethanolamine, but when cultured in nonsupplemented medium, they produce high frequencies of abnormally shaped cells as well as cells exhibiting severe septation defects, including multiple, mispositioned, deformed, and misoriented septa. Our results demonstrate that phosphatidylethanolamine is essential for cell growth and for normal cytokinesis and cellular morphogenesis in S. pombe, and they illustrate the usefulness of this model eukaryote for investigating potentially conserved biological and molecular functions of phosphatidylethanolamine.
为了研究磷脂酰乙醇胺对裂殖酵母粟酒裂殖酵母生长和形态发生的贡献,我们对该生物体中的三个预测基因进行了表征,分别命名为psd1、psd2和psd3,它们编码磷脂酰丝氨酸脱羧酶,该酶在真核生物和原核生物中均催化磷脂酰丝氨酸向磷脂酰乙醇胺的转化。在任何一个或两个psd基因中携带缺失的粟酒裂殖酵母突变体在复杂丰富培养基和合成限定基本培养基中都能存活。然而,在所有三个psd基因中携带缺失的突变体(psd1-3Δ突变体)在丰富培养基中生长缓慢,在基本培养基中无法存活,这表明psd1至psd3基因产物具有重叠的基本细胞功能。用乙醇胺补充生长培养基,乙醇胺可通过肯尼迪途径转化为磷脂酰乙醇胺,这可使psd1-3Δ细胞在基本培养基中恢复生长,表明磷脂酰乙醇胺对粟酒裂殖酵母细胞生长至关重要。即使在补充了乙醇胺的培养基中,psd1-3Δ细胞产生的磷脂酰乙醇胺水平也低于野生型细胞,这表明肯尼迪途径只能部分补偿粟酒裂殖酵母中磷脂酰丝氨酸脱羧酶活性的丧失。在补充了乙醇胺的培养基中,psd1-3Δ细胞在形态上似乎与野生型粟酒裂殖酵母细胞没有区别,但在未补充培养基中培养时,它们会产生高频率的异常形状细胞以及表现出严重隔膜缺陷的细胞,包括多个、位置错误、变形和方向错误的隔膜。我们的结果表明,磷脂酰乙醇胺对粟酒裂殖酵母的细胞生长、正常胞质分裂和细胞形态发生至关重要,并且它们说明了这种模式真核生物在研究磷脂酰乙醇胺潜在保守的生物学和分子功能方面的有用性。