Pechter Kieran B, Gallagher Larry, Pyles Harley, Manoil Colin S, Harwood Caroline S
Department of Microbiology, University of Washington, Seattle, Washington, USA.
Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
J Bacteriol. 2015 Dec 28;198(5):867-76. doi: 10.1128/JB.00771-15.
Rhodopseudomonas palustris is an alphaproteobacterium that has served as a model organism for studies of photophosphorylation, regulation of nitrogen fixation, production of hydrogen as a biofuel, and anaerobic degradation of aromatic compounds. This bacterium is able to transition between anaerobic photoautotrophic growth, anaerobic photoheterotrophic growth, and aerobic heterotrophic growth. As a starting point to explore the genetic basis for the metabolic versatility of R. palustris, we used transposon mutagenesis and Tn-seq to identify 552 genes as essential for viability in cells growing aerobically on semirich medium. Of these, 323 have essential gene homologs in the alphaproteobacterium Caulobacter crescentus, and 187 have essential gene homologs in Escherichia coli. There were 24 R. palustris genes that were essential for viability under aerobic growth conditions that have low sequence identity but are likely to be functionally homologous to essential E. coli genes. As expected, certain functional categories of essential genes were highly conserved among the three organisms, including translation, ribosome structure and biogenesis, secretion, and lipid metabolism. R. palustris cells divide by budding in which a sessile cell gives rise to a motile swarmer cell. Conserved cell cycle genes required for this developmental process were essential in both C. crescentus and R. palustris. Our results suggest that despite vast differences in lifestyles, members of the alphaproteobacteria have a common set of essential genes that is specific to this group and distinct from that of gammaproteobacteria like E. coli.
Essential genes in bacteria and other organisms are those absolutely required for viability. Rhodopseudomonas palustris has served as a model organism for studies of anaerobic aromatic compound degradation, hydrogen gas production, nitrogen fixation, and photosynthesis. We used the technique of Tn-seq to determine the essential genes of R. palustris grown under heterotrophic aerobic conditions. The transposon library generated in this study will be useful for future studies to identify R. palustris genes essential for viability under specialized growth conditions and also for survival under conditions of stress.
沼泽红假单胞菌是一种α-变形菌,已成为研究光合磷酸化、固氮调节、作为生物燃料的氢气生产以及芳香化合物厌氧降解的模式生物。这种细菌能够在厌氧光合自养生长、厌氧光合异养生长和好氧异养生长之间转换。作为探索沼泽红假单胞菌代谢多功能性遗传基础的起点,我们使用转座子诱变和Tn-seq来鉴定552个基因,这些基因对于在半丰富培养基上有氧生长的细胞的生存能力至关重要。其中,323个基因在α-变形菌新月柄杆菌中有必需基因同源物,187个基因在大肠杆菌中有必需基因同源物。有24个沼泽红假单胞菌基因在有氧生长条件下对生存能力至关重要,它们的序列同一性较低,但可能与大肠杆菌的必需基因在功能上同源。正如预期的那样,必需基因的某些功能类别在这三种生物中高度保守,包括翻译、核糖体结构和生物发生、分泌和脂质代谢。沼泽红假单胞菌细胞通过出芽进行分裂,其中一个固着细胞产生一个游动的游动细胞。这个发育过程所需的保守细胞周期基因在新月柄杆菌和沼泽红假单胞菌中都是必需的。我们的结果表明,尽管生活方式存在巨大差异,但α-变形菌成员有一组共同的必需基因,这组基因是该类群特有的,与大肠杆菌等γ-变形菌不同。
细菌和其他生物中的必需基因是生存绝对必需的基因。沼泽红假单胞菌已成为研究厌氧芳香化合物降解、氢气生产、固氮和光合作用的模式生物。我们使用Tn-seq技术来确定在异养有氧条件下生长的沼泽红假单胞菌的必需基因。本研究中产生的转座子文库将有助于未来的研究,以鉴定沼泽红假单胞菌在特殊生长条件下生存以及在应激条件下存活所必需的基因。