Bhat Swapna, Boynton Tye O, Pham Dan, Shimkets Lawrence J
Department of Microbiology, University of Georgia, Athens, Georgia, United States of America.
PLoS One. 2014 Jun 6;9(6):e99622. doi: 10.1371/journal.pone.0099622. eCollection 2014.
Myxococcus xanthus responds to amino acid limitation by producing fruiting bodies containing dormant spores. During development, cells produce triacylglycerides in lipid bodies that become consumed during spore maturation. As the cells are starved to induce development, the production of triglycerides represents a counterintuitive metabolic switch. In this paper, lipid bodies were quantified in wild-type strain DK1622 and 33 developmental mutants at the cellular level by measuring the cross sectional area of the cell stained with the lipophilic dye Nile red. We provide five lines of evidence that triacylglycerides are derived from membrane phospholipids as cells shorten in length and then differentiate into myxospores. First, in wild type cells, lipid bodies appear early in development and their size increases concurrent with an 87% decline in membrane surface area. Second, developmental mutants blocked at different stages of shortening and differentiation accumulated lipid bodies proportionate with their cell length with a Pearson's correlation coefficient of 0.76. Third, peripheral rods, developing cells that do not produce lipid bodies, fail to shorten. Fourth, genes for fatty acid synthesis are down-regulated while genes for fatty acid degradation are up regulated. Finally, direct movement of fatty acids from membrane lipids in growing cells to lipid bodies in developing cells was observed by pulse labeling cells with palmitate. Recycling of lipids released by Programmed Cell Death appears not to be necessary for lipid body production as a fadL mutant was defective in fatty acid uptake but proficient in lipid body production. The lipid body regulon involves many developmental genes that are not specifically involved in fatty acid synthesis or degradation. MazF RNA interferase and its target, enhancer-binding protein Nla6, appear to negatively regulate cell shortening and TAG accumulation whereas most cell-cell signals activate these processes.
黄色粘球菌通过产生含有休眠孢子的子实体来应对氨基酸限制。在发育过程中,细胞在脂质体中产生三酰甘油,这些三酰甘油在孢子成熟过程中被消耗。当细胞饥饿以诱导发育时,三酰甘油的产生代表了一种违反直觉的代谢转换。在本文中,通过测量用亲脂性染料尼罗红染色的细胞的横截面积,在细胞水平上对野生型菌株DK1622和33个发育突变体中的脂质体进行了定量。我们提供了五条证据,表明随着细胞长度缩短并分化为粘孢子,三酰甘油源自膜磷脂。第一,在野生型细胞中,脂质体在发育早期出现,其大小增加,同时膜表面积下降87%。第二,在缩短和分化的不同阶段受阻的发育突变体积累的脂质体与其细胞长度成比例,皮尔逊相关系数为0.76。第三,外周杆状细胞,即不产生脂质体的发育细胞,不会缩短。第四,脂肪酸合成基因被下调,而脂肪酸降解基因被上调。最后,通过用棕榈酸脉冲标记细胞,观察到脂肪酸从生长细胞中的膜脂直接转移到发育细胞中的脂质体。程序性细胞死亡释放的脂质的再循环似乎不是脂质体产生所必需的,因为fadL突变体在脂肪酸摄取方面有缺陷,但在脂质体产生方面表现正常。脂质体调控子涉及许多并非专门参与脂肪酸合成或降解的发育基因。MazF RNA干扰酶及其靶标增强子结合蛋白Nla6似乎对细胞缩短和TAG积累起负调控作用,而大多数细胞间信号激活这些过程。