Kós Péter B, Deák Zsuzsanna, Cheregi Otilia, Vass Imre
Institute of Plant Biology, Biological Research Center, Szeged, Hungary.
Biochim Biophys Acta. 2008 Jan;1777(1):74-83. doi: 10.1016/j.bbabio.2007.10.015. Epub 2007 Nov 12.
In Thermosynechococcus elongatus BP-1, which is the preferred organism in recent structural studies of PSII, three psbA and two psbD genes code for three D1 and one D2 protein isoforms, respectively. The regulation and function of these genes and protein products is largely unknown. Therefore, we used quantitative RT-PCR to follow changes in the mRNA level of the respective genes, in combination with biophysical measurements to detect changes in the electron transport activity of Photosystem II under exposure to different visible and UV light, and temperature conditions. In cells which are acclimated to 40 micromol m(-2)s(-1) growth light conditions at 40 degrees C the main populations of the psbA and psbD transcripts arise from the psbA1 and psbD1 genes, respectively. When the temperature is raised to 60 degrees C psbA1 becomes the single dominating psbA mRNA species. Upon exposure of the cells to 500 micromol m(-2)s(-1) intensity visible light psbA3 replaces psbA1 as the dominating psbA mRNA species, and psbD2 increases at the expense of psbD1. UV-B radiation also increases the abundance of psbA3, and psbD2 at the expense of psbA1 and psbD1, respectively. From the different extent of total D1 protein loss in the absence and presence of lincomycin it was estimated that the PsbA3 protein isoform replaces PsbA1 in about 65% of PSII centers after 2 h of high light acclimation. Under the conditions of different psbA transcript distributions chlorophyll fluorescence and thermoluminescence measurements were applied to monitor charge recombination characteristics of the S2Q(A)(-) and S2Q(B)(-) states. We obtained faster decay of flash-induced chlorophyll fluorescence in the presence of DCMU, as well as lower peak temperature of the Q and B thermoluminescence bands when PsbA3 replaced PsbA1 as the main D1 protein isoform. The relevance of dynamic changes in the abundance of psbA and psbD transcript levels, as well as D1 protein isoforms in the acclimation of T. elongatus to changing environmental conditions is discussed.
在嗜热栖热菌BP - 1(近期PSII结构研究中常用的生物体)中,三个psbA基因和两个psbD基因分别编码三种D1蛋白异构体和一种D2蛋白异构体。这些基因及其蛋白质产物的调控和功能在很大程度上尚不清楚。因此,我们使用定量RT - PCR来跟踪各个基因mRNA水平的变化,并结合生物物理测量来检测在不同可见光、紫外光和温度条件下光系统II电子传递活性的变化。在适应40℃、40 μmol m⁻² s⁻¹生长光照条件的细胞中,psbA和psbD转录本的主要群体分别来自psbA1和psbD1基因。当温度升至60℃时,psbA1成为唯一占主导地位的psbA mRNA种类。当细胞暴露于强度为500 μmol m⁻² s⁻¹的可见光下时,psbA3取代psbA1成为占主导地位的psbA mRNA种类,并且psbD2增加,psbD1减少。UV - B辐射也分别以psbA1和psbD1为代价增加了psbA3和psbD2的丰度。根据在有无林可霉素存在的情况下总D1蛋白损失的不同程度,估计在高光适应2小时后,约65%的PSII中心中PsbA3蛋白异构体取代了PsbA1。在不同psbA转录本分布的条件下,应用叶绿素荧光和热发光测量来监测S2Q(A)⁻和S2Q(B)⁻状态的电荷复合特性。当PsbA3取代PsbA1作为主要的D1蛋白异构体时,我们在DCMU存在的情况下获得了更快衰减的闪光诱导叶绿素荧光,以及Q和B热发光带的更低峰值温度。本文讨论了psbA和psbD转录本水平以及D1蛋白异构体丰度的动态变化在嗜热栖热菌适应不断变化的环境条件中的相关性。