Rhodopseudomonas palustris CGA009 has two functional ppsR genes, each of which encodes a repressor of photosynthesis gene expression

Abstract

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.