From the Department of Biochemistry and Molecular Biology and.
From the Department of Biochemistry and Molecular Biology and the Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802 and.
J Biol Chem. 2014 Jun 6;289(23):15904-14. doi: 10.1074/jbc.M114.555631. Epub 2014 Apr 24.
The biogenesis of thylakoid membranes in cyanobacteria is presently not well understood, but the vipp1 gene product has been suggested to play an important role in this process. Previous studies in Synechocystis sp. PCC 6803 reported that vipp1 (sll0617) was essential. By constructing a fully segregated null mutant in vipp1 (SynPCC7002_A0294) in Synechococcus sp. PCC 7002, we show that Vipp1 is not essential. Spectroscopic studies revealed that Photosystem I (PS I) was below detection limits in the vipp1 mutant, but Photosystem II (PS II) was still assembled and was active. Thylakoid membranes were still observed in vipp1 mutant cells and resembled those in a psaAB mutant that completely lacks PS I. When the vipp1 mutation was complemented with the orthologous vipp1 gene from Synechocystis sp. PCC 6803 that was expressed from the strong P(cpcBA) promoter, PS I content and activities were restored to normal levels, and cells again produced thylakoids that were indistinguishable from those of wild type. Transcription profiling showed that psaAB transcripts were lower in abundance in the vipp1 mutant. However, when the yfp gene was expressed from the P(psaAB) promoter in the presence and the absence of Vipp1, no difference in YFP expression was observed, which shows that Vipp1 is not a transcription factor for the psaAB genes. This study shows that thylakoids are still produced in the absence of Vipp1 and that normal thylakoid biogenesis in Synechococcus sp. PCC 7002 requires expression and biogenesis of PS I, which in turn requires Vipp1.
蓝藻中类囊体膜的生物发生目前还不太清楚,但 vipp1 基因产物被认为在这个过程中发挥着重要作用。先前在集胞藻 PCC 6803 中的研究报告称,vipp1(sll0617)是必需的。通过构建 Synechococcus sp. PCC 7002 中 vipp1(SynPCC7002_A0294)的完全分离的缺失突变体,我们表明 Vipp1 不是必需的。光谱研究表明,vipp1 突变体中的光系统 I(PSI)低于检测限,但光系统 II(PS II)仍然组装并具有活性。在 vipp1 突变体细胞中仍观察到类囊体膜,并且类似于完全缺乏 PS I 的 psaAB 突变体。当用来自集胞藻 PCC 6803 的同源 vipp1 基因(由强 P(cpcBA)启动子表达)对 vipp1 突变进行互补时,PSI 含量和活性恢复到正常水平,并且细胞再次产生与野生型无异的类囊体。转录谱分析表明,vipp1 突变体中 psaAB 转录物的丰度较低。然而,当 yfp 基因在存在和不存在 Vipp1 的情况下由 P(psaAB)启动子表达时,观察到 YFP 表达没有差异,这表明 Vipp1 不是 psaAB 基因的转录因子。这项研究表明,在缺乏 Vipp1 的情况下仍能产生类囊体,并且在 Synechococcus sp. PCC 7002 中正常的类囊体生物发生需要 PS I 的表达和生物发生,而这又需要 Vipp1。