Mukhopadhyay B, Johnson E F, Wolfe R S
Department of Microbiology, University of Illinois at Urbana-Champaign, B103 Chemical and Life Sciences Laboratory, 601 South Goodwin Avenue, Urbana, IL 61801, USA.
Proc Natl Acad Sci U S A. 2000 Oct 10;97(21):11522-7. doi: 10.1073/pnas.97.21.11522.
The methanarchaeon, Methanococcus jannaschii, a hyperthermophilic, autotrophic, and strictly hydrogenotrophic inhabitant of submarine hydrothermal vents, was cultivated in a reactor at two hydrogen partial pressure (p(H(2))) values, 178 kPa (high) and 650 Pa (ultralow), and the cells were subjected to a comparative proteome analysis. From these studies, it was discovered that, when p(H(2)) was high and the cell density was low (a combination representing a hydrogen-excess condition), the cells possessed very low or undetectable levels of four flagella-related polypeptides (FlaB2, FlaB3, FlaD, and FlaE); electron microscopic examination showed that most of these cells were devoid of flagella. Flagella synthesis occurred when hydrogen became limiting either at high cell density under high p(H(2)) or at low cell density under low p(H(2)). The results from a p(H(2))-shift experiment corroborated the above observations. The p(H(2))-dependent changes in the levels of two methanogenic enzymes (MTD and HMDX) were as expected, and thus they served as internal controls. To our knowledge, this is the first example for the regulation of expression of flagella by hydrogen in any domain of life and for a control of any kind on flagella synthesis in the archaea. Our work also provides the only known example for each of the following: (i) the pure culture cultivation of a methanogen at an ultralow, near ecologically relevant p(H(2)); (ii) experimental functional genomics for M. jannaschii; and (iii) the use of proteomics with M. jannaschii.
甲烷古菌詹氏甲烷球菌是一种嗜热、自养且严格以氢为营养源的海底热液喷口栖息生物,在一个反应器中于两个氢气分压(p(H₂))值下进行培养,分别为178 kPa(高)和650 Pa(超低),并对细胞进行了比较蛋白质组分析。通过这些研究发现,当p(H₂)高且细胞密度低(这种组合代表氢气过剩条件)时,细胞中四种与鞭毛相关的多肽(FlaB2、FlaB3、FlaD和FlaE)水平极低或检测不到;电子显微镜检查表明,这些细胞中的大多数没有鞭毛。当氢气在高p(H₂)下细胞密度高或低p(H₂)下细胞密度低时变得有限时,鞭毛合成发生。p(H₂)转换实验的结果证实了上述观察结果。两种产甲烷酶(MTD和HMDX)水平的p(H₂)依赖性变化如预期,因此它们作为内部对照。据我们所知,这是生命任何领域中氢气对鞭毛表达进行调控以及古菌中对鞭毛合成进行任何形式控制的首个例子。我们的工作还为以下各项提供了唯一已知的例子:(i)在超低、接近生态相关的p(H₂)下对产甲烷菌进行纯培养;(ii)詹氏甲烷球菌的实验功能基因组学;以及(iii)对詹氏甲烷球菌使用蛋白质组学。