Colón-López M S, Sherman D M, Sherman L A
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA.
J Bacteriol. 1997 Jul;179(13):4319-27. doi: 10.1128/jb.179.13.4319-4327.1997.
Cyanothece sp. strain ATCC 51142 is a unicellular, diazotrophic cyanobacterium which demonstrated extensive metabolic periodicities of photosynthesis, respiration, and nitrogen fixation when grown under N2-fixing conditions. N2 fixation and respiration peaked at 24-h intervals early in the dark or subjective-dark period, whereas photosynthesis was approximately 12 h out of phase and peaked toward the end of the light or subjective-light phase. Gene regulation studies demonstrated that nitrogenase is carefully controlled at the transcriptional and posttranslational levels. Indeed, Cyanothece sp. strain ATCC 51142 has developed an expensive mode of regulation, such that nitrogenase was synthesized and degraded each day. These patterns were seen when cells were grown under either light-dark or continuous-light conditions. Nitrogenase mRNA was synthesized from the nifHDK operon during the first 4 h of the dark period under light-dark conditions or during the first 6 h of the subjective-dark period when grown in continuous light. The nitrogenase NifH and NifDK subunits reached a maximum level at 4 to 10 h in the dark or subjective-dark periods and were shown by Western blotting and electron microscopy immunocytochemistry to be thoroughly degraded toward the end of the dark periods. An exception is the NifDK protein (MoFe-protein), which appeared not to be completely degraded under continuous-light conditions. We hypothesize that cellular O2 levels were kept low by decreasing photosynthesis and by increasing respiration in the early dark or subjective-dark periods to permit nitrogenase activity. The subsequent increase in O2 levels resulted in nitrogenase damage and eventual degradation.
蓝细菌菌株ATCC 51142是一种单细胞、固氮蓝细菌,在固氮条件下生长时,其光合作用、呼吸作用和固氮作用表现出广泛的代谢周期性。固氮作用和呼吸作用在黑暗或主观黑暗期早期以24小时为间隔达到峰值,而光合作用则大约相差12小时,在光照或主观光照期结束时达到峰值。基因调控研究表明,固氮酶在转录和翻译后水平受到严格控制。事实上,蓝细菌菌株ATCC 51142已经形成了一种复杂的调控模式,即固氮酶每天合成和降解。当细胞在明暗或连续光照条件下生长时,都能观察到这些模式。在明暗条件下的黑暗期的前4小时或连续光照下生长时主观黑暗期的前6小时,固氮酶mRNA从nifHDK操纵子合成。固氮酶NifH和NifDK亚基在黑暗或主观黑暗期的4至10小时达到最高水平,蛋白质免疫印迹和电子显微镜免疫细胞化学显示,在黑暗期结束时它们被彻底降解。一个例外是NifDK蛋白(钼铁蛋白),在连续光照条件下它似乎没有被完全降解。我们推测,通过在黑暗或主观黑暗期早期降低光合作用和增加呼吸作用,使细胞内的氧气水平保持较低,以允许固氮酶发挥活性。随后氧气水平的升高导致固氮酶受损并最终降解。