Kulkarni R D, Schaefer M R, Golden S S
Department of Biology, Texas A&M University, College Station 77843-3258.
J Bacteriol. 1992 Jun;174(11):3775-81. doi: 10.1128/jb.174.11.3775-3781.1992.
The psbA genes, which encode the D1 protein of photosystem II, constitute a multigene family in the cyanobacterium Synechococcus sp. strain PCC 7942. Levels of messages from the three psbA genes change rapidly when cells are shifted from low-light to high-light conditions: the psbAI message level drops, whereas psbAII and psbAIII message levels increase dramatically. We examined the potential contributions of transcriptional and posttranscriptional processes in these high-light responses by subjecting cells that had been grown in a turbidostat at a standard light intensity (130 microeinsteins [microE] m-2 s-1) to either the same or a higher light intensity (500 microE m-2 s-1) in the presence or absence of rifampin. Northern (RNA blot) analysis of RNA isolated from cells subjected to high light showed that the increases in psbAII and psbAIII transcripts were blocked by rifampin. This suggests a transcriptional induction of these genes at high light intensities. Increased mRNA stability does not contribute to their accumulation in high-light conditions, since their half-life values did not increase relative to the half-lives measured at the standard light intensity. The rate of disappearance of the psbAI transcript in cells shifted to high light was diminished when either transcription or translation was blocked by rifampin or chloramphenicol, suggesting that accelerated degradation of the message requires de novo synthesis of a protein factor. When rifampin was added 10 min after the shift to high light intensity rather than before the shift, psbAI and psbAIII messages, but not the psbAII message, decayed at a faster rate. Susceptibility of the psbAIII transcript to the high-light-induced factor was also demonstrated by addition of chloramphenicol prior to the shaft to high light. psbAIII transcript levels went up more than twofold higher in chloramphenicol-treated cells than in untreated cells, whereas psbAII transcript levels were affected by the inhibitor. These experiments provide evidence that either new or increased synthesis of a degradation factor which affects a subset of Synechococcus transcripts occurs in cells subjected to high light intensity.
编码光系统II的D1蛋白的psbA基因,在聚球藻属蓝细菌PCC 7942菌株中构成一个多基因家族。当细胞从低光照条件转移到高光照条件时,来自三个psbA基因的信使水平迅速变化:psbAI信使水平下降,而psbAII和psbAIII信使水平急剧增加。我们通过使在标准光照强度(130微爱因斯坦[μE] m-2 s-1)下在恒浊器中生长的细胞在有或没有利福平的情况下暴露于相同或更高的光照强度(500 μE m-2 s-1),来研究转录和转录后过程在这些高光反应中的潜在作用。对从高光处理的细胞中分离的RNA进行Northern(RNA印迹)分析表明,psbAII和psbAIII转录本的增加被利福平阻断。这表明在高光强度下这些基因存在转录诱导。mRNA稳定性的增加并不导致它们在高光条件下的积累,因为它们的半衰期值相对于在标准光照强度下测量的半衰期没有增加。当转录或翻译被利福平或氯霉素阻断时,转移到高光的细胞中psbAI转录本的消失速率降低,这表明信使的加速降解需要从头合成一种蛋白质因子。当在转移到高光强度后10分钟而不是转移前添加利福平,psbAI和psbAIII信使,但不是psbAII信使,以更快的速率衰减。在转移到高光之前添加氯霉素也证明了psbAIII转录本对高光诱导因子的敏感性。氯霉素处理的细胞中psbAIII转录本水平比未处理的细胞高出两倍多,而psbAII转录本水平受到抑制剂的影响。这些实验提供了证据,即在高光强度下的细胞中发生了影响聚球藻转录本子集的降解因子的新合成或合成增加。