Soitamo A J, Sippola K, Aro E M
Department of Biology, University of Turku, BioCity A, Finland.
Plant Mol Biol. 1998 Aug;37(6):1023-33. doi: 10.1023/a:1006077824075.
Expression of the psbA genes, which in the cyanobacterium Synechococcus sp. PCC 7942 encode two different forms of the reaction centre D1 protein of photosystem II (D1:1 and D1:2), was studied under different light and temperature conditions. In addition to the mature 1200 nt psbA messages, three shorter mRNA fragments of 220, 320 and 900 nt were also found. All three mRNA fragments could be recognized by using different gene probes from the coding region of the psbAI gene, whereas the corresponding psbAII/III gene probes recognized only the 220 nt mRNA fragment. The 5' 320 nt mRNA fragment from the psbAI gene probably represents a degradation product, since the corresponding 3' 900 nt psbAI mRNA fragment was also detected. By contrast, the 5' 220 nt mRNA fragment of all psbA messages is suggested to be a truncated psbA transcript, since no corresponding 3' fragment was ever found. Inhibition of translation either by a protein synthesis inhibitor or by a shift of cells to lower temperature, increased the number of 1200 nt psbAII/III messages but the number of 5' 220 nt psbAII/III mRNA fragment increased even more dramatically. The first 66 bp after ATG, where the psbAI and psbAII/III genes mostly differ from each other, also appeared important in determining the amount of produced truncated psbA transcripts, as evidenced by the expression of different tac-psbA constructs in the presence of protein synthesis inhibitor. We suggest that both the psbAI and the psbAII/III genes have a latent intragenic termination site and truncated psbA transcripts are produced at high levels under stress conditions when transcription becomes uncoupled from translation. This is to prevent wasting metabolic energy in the production of unused transcripts.
研究了集胞藻属蓝细菌PCC 7942中psbA基因的表达情况,该基因编码光系统II反应中心D1蛋白的两种不同形式(D1:1和D1:2),研究是在不同光照和温度条件下进行的。除了成熟的1200 nt psbA信使核糖核酸外,还发现了三个较短的mRNA片段,长度分别为220、320和900 nt。使用来自psbAI基因编码区的不同基因探针可以识别所有这三个mRNA片段,而相应的psbAII/III基因探针仅识别220 nt的mRNA片段。psbAI基因的5' 320 nt mRNA片段可能代表一种降解产物,因为也检测到了相应的3' 900 nt psbAI mRNA片段。相比之下,所有psbA信使核糖核酸的5' 220 nt mRNA片段被认为是一种截短的psbA转录本,因为从未发现相应的3'片段。无论是用蛋白质合成抑制剂抑制翻译,还是将细胞转移到较低温度,都会增加1200 nt psbAII/III信使核糖核酸的数量,但5' 220 nt psbAII/III mRNA片段的数量增加得更为显著。ATG之后的前66个碱基对,psbAI和psbAII/III基因在此处大多彼此不同,在确定截短的psbA转录本的产生量方面似乎也很重要,这在蛋白质合成抑制剂存在的情况下不同tac-psbA构建体的表达中得到了证明。我们认为,psbAI和psbAII/III基因都有一个潜在的基因内终止位点,并且在应激条件下,当转录与翻译解偶联时,截短的psbA转录本会大量产生。这是为了防止在产生未使用的转录本时浪费代谢能量。