Tabei Yosuke, Okada Katsuhiko, Makita Nobuaki, Tsuzuki Mikio
School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Horinouchi, Hachioji, Tokyo, Japan.
FEBS J. 2009 Jan;276(1):187-98. doi: 10.1111/j.1742-4658.2008.06772.x. Epub 2008 Nov 19.
Synechocystis sp. PCC 6803 exhibits light-activated heterotrophic growth (LAHG) under dark conditions with glucose as a carbon source. The light activation is remarkable at a late period of photoautotrophic preculture, such as the late-linear and stationary growth phases. To understand the physiological effects of light irradiation and glucose under LAHG conditions, their effects on the expression of soluble proteins were analyzed by means of 2D-PAGE. Various soluble proteins, which were minimal under photoautotrophic preculture conditions, were observed clearly under LAHG conditions, suggesting that proteins were synthesized actively under these conditions. Fructose 1,6-bisphosphate aldolase, one of the glycolytic enzymes, was found to be induced under LAHG conditions on 2D-PAGE. The activity of fructose 1,6-bisphosphate aldolase, which had decreased during photoautotrophic preculture, also increased under LAHG conditions, similar to the mRNA level of the encoding gene, fbaA. In addition, we found that a deletion mutant of sll1330, a putative gene containing a helix-turn-helix DNA-binding motif, could not grow under LAHG conditions, whereas it could grow photoautotrophically. The increases in the protein level of FbaA and fbaA gene expression observed in wild-type cells under LAHG conditions were greatly inhibited in the deletion mutant. These results suggest that the regulation of fbaA gene expression by way of sll1330 is one of the important processes in Synechocystis sp. PCC 6803 under light pulse LAHG conditions.
聚球藻属PCC 6803在黑暗条件下以葡萄糖作为碳源时表现出光激活异养生长(LAHG)。在光合自养预培养的后期,如线性生长后期和稳定期,光激活现象非常显著。为了了解LAHG条件下光照和葡萄糖的生理作用,通过二维聚丙烯酰胺凝胶电泳(2D-PAGE)分析了它们对可溶性蛋白质表达的影响。在光合自养预培养条件下含量极少的各种可溶性蛋白质,在LAHG条件下清晰可见,这表明在这些条件下蛋白质能够活跃合成。糖酵解酶之一的果糖-1,6-二磷酸醛缩酶在2D-PAGE上被发现在LAHG条件下被诱导。在光合自养预培养过程中活性降低的果糖-1,6-二磷酸醛缩酶的活性,在LAHG条件下也有所增加,这与编码基因fbaA的mRNA水平相似。此外,我们发现一个假定的含有螺旋-转角-螺旋DNA结合基序的基因sll1330的缺失突变体在LAHG条件下无法生长,而它能够进行光合自养生长。在野生型细胞中LAHG条件下观察到的FbaA蛋白水平和fbaA基因表达的增加在缺失突变体中受到极大抑制。这些结果表明,通过sll1330对fbaA基因表达的调控是聚球藻属PCC 6803在光脉冲LAHG条件下的重要过程之一。