Braatsch Stephan, Bernstein Jeffrey R, Lessner Faith, Morgan Jennifer, Liao James C, Harwood Caroline S, Beatty J Thomas
Department of Microbiology and Immunology, University of British Columbia, 2350 Health Sciences Mall, Vancouver, British Columbia V6T 1Z3, Canada.
Biochemistry. 2006 Dec 5;45(48):14441-51. doi: 10.1021/bi061074b.
The PpsR protein is a regulator of redox-dependent photosystem development in purple phototrophic bacteria. In contrast to most species, Rhodopseudomonas palustris contains two ppsR genes. We show that the inactivation of each of the R. palustris strain CGA009 ppsR genes results in an elevated level of formation of the photosystem under dark aerobic conditions. Absorption spectra of the two PpsR mutants revealed qualitative and quantitative differences in light-harvesting peak amplitude increases. A sequence difference in the helix-turn-helix DNA binding motif of PpsR2 (Arg 439 to Cys) between R. palustris strains CEA001 and CGA009 is shown to be a natural polymorphism that does not inactivate the repressor activity of the protein. To evaluate which photosynthesis genes are regulated by the two PpsR proteins, transcriptome profiles of the CGA009 and PpsR mutant strains were analyzed in microarray experiments. Transcription of most but not all photosystem genes was derepressed in the mutant strains to levels consistent with the in vivo absorption spectra, mathematical analyses of peak shapes and amplitudes, reaction center protein levels, and real-time PCR of selected mRNAs. Closely spaced PpsR binding motif repeats were identified 5' of genes that were derepressed in the transcriptome analysis of PpsR mutants. This work shows that both the PpsR1 and PpsR2 proteins from R. palustris strain CGA009 function as oxygen-responsive transcriptional repressors.
PpsR蛋白是紫色光合细菌中氧化还原依赖性光系统发育的调节因子。与大多数物种不同,沼泽红假单胞菌含有两个ppsR基因。我们发现,沼泽红假单胞菌CGA009菌株的每个ppsR基因失活都会导致在黑暗需氧条件下光系统形成水平升高。两个PpsR突变体的吸收光谱显示,在捕光峰幅度增加方面存在定性和定量差异。沼泽红假单胞菌CEA001和CGA009菌株之间PpsR2(从Arg 439到Cys)的螺旋-转角-螺旋DNA结合基序中的序列差异被证明是一种自然多态性,不会使该蛋白的阻遏活性失活。为了评估哪些光合作用基因受这两种PpsR蛋白调控,在微阵列实验中分析了CGA009和PpsR突变体菌株的转录组谱。在突变体菌株中,大多数但不是所有光系统基因的转录被解除抑制,达到与体内吸收光谱、峰形和幅度的数学分析、反应中心蛋白水平以及选定mRNA的实时PCR一致的水平。在PpsR突变体转录组分析中被解除抑制的基因的5'端,鉴定出了紧密间隔的PpsR结合基序重复序列。这项工作表明,沼泽红假单胞菌CGA009菌株的PpsR1和PpsR2蛋白均作为氧响应转录阻遏物发挥作用。