ABSTRACT: An integrative genomics approach to explore diversity of light harvesting antenna complexes from environmental isolates of the purple non-sulfur bacterium Rhodopseudomonas palustris
Project description:Peripheral light harvesting (LH) antenna complexes have been studied extensively in the purple nonsulfur bacterium Rhodopseudomonas palustris because it produces different types of LH complexes under high light intensities (LH2 complex) and low light intensities (LH3 and LH4 complex). The ability of R. palustris to alter its peripheral LH complexes in response to changes in light intensity is attributed to the multiple operons that encode the a and b peptides that make up these complexes, whose expression is affected by light intensity, light quality, and oxygen tension. However, low resolution structures, amino acid similarities between the complexes, and a lack of transcriptional analysis made it difficult to determine the LH complexes composition and functions under different light intensities. It was also unclear how much diversity of the R. palustris LH complexes exists in nature.Results: To gain insight into the composition of the LH complexes, their function under high light intensities and low light intensities, and their prevalence in the environment we undertook an integrative genomics approach using 15 closely related R. palustris strains isolated from the environment and 5 R. palustris ecotypes whose genomes have been sequenced. We sequenced the genomes for the 15 closely related strains and using RNA-seq carried out transcriptomic analysis on all 20 strains grown under high light intensity and low light intensity. We were able to determine that even closely related R. palustris strains had differences in their pucBA gene content and expression, even under the same growth conditions. We also found that the LH2 complex could compensate for the lack of an LH4 complex under LL intensities but not under extremely LL intensities. Conclusions: This is the first time an integrative genomics approach has been used to study light harvesting in the environment. The variation observed in LH gene composition and expression in environmental isolates of R. palustris likely reflects how these strains have adapted to specific light conditions in the environment. We have also shown that there is redundancy between some of the LH complexes under certain light intensities, which may partially explain why multiple operons encoding LH complexes have evolved and been maintained in R. palustris. Examing the variation observed in LH gene composition and expression in various environmental isolates
Project description:Peripheral light harvesting (LH) antenna complexes have been studied extensively in the purple nonsulfur bacterium Rhodopseudomonas palustris because it produces different types of LH complexes under high light intensities (LH2 complex) and low light intensities (LH3 and LH4 complex). The ability of R. palustris to alter its peripheral LH complexes in response to changes in light intensity is attributed to the multiple operons that encode the a and b peptides that make up these complexes, whose expression is affected by light intensity, light quality, and oxygen tension. However, low resolution structures, amino acid similarities between the complexes, and a lack of transcriptional analysis made it difficult to determine the LH complexes composition and functions under different light intensities. It was also unclear how much diversity of the R. palustris LH complexes exists in nature.Results: To gain insight into the composition of the LH complexes, their function under high light intensities and low light intensities, and their prevalence in the environment we undertook an integrative genomics approach using 15 closely related R. palustris strains isolated from the environment and 5 R. palustris ecotypes whose genomes have been sequenced. We sequenced the genomes for the 15 closely related strains and using RNA-seq carried out transcriptomic analysis on all 20 strains grown under high light intensity and low light intensity. We were able to determine that even closely related R. palustris strains had differences in their pucBA gene content and expression, even under the same growth conditions. We also found that the LH2 complex could compensate for the lack of an LH4 complex under LL intensities but not under extremely LL intensities. Conclusions: This is the first time an integrative genomics approach has been used to study light harvesting in the environment. The variation observed in LH gene composition and expression in environmental isolates of R. palustris likely reflects how these strains have adapted to specific light conditions in the environment. We have also shown that there is redundancy between some of the LH complexes under certain light intensities, which may partially explain why multiple operons encoding LH complexes have evolved and been maintained in R. palustris.
Project description:The purple bacterium Rhodopseudomonas palustris is a model organism for dissecting the energy and electron transfer processes that have evolved in phototrophic organisms. This bacterium is of particular interest because, in addition to driving its metabolism via solar energy capture, it is capable of nitrogen and carbon dioxide fixation, producing hydrogen and utilising a wide range of organic compounds. Understanding these processes underpins the potential exploitation of Rhodopseudomonas palustris for synthetic biology, biohydrogen production and bioremediation, for example. Like other purple bacteria, Rhodopseudomonas palustris has 2 light-harvesting (LH) systems: LH1 and LH2. The former has already been extensively characterised by X-ray crystallography and cryo-EM. The aim of this proteomics project is to provide complementary information to support the cryo-EM mapping of LH2 structure.
Project description:Rhodopseudomonas palustris is a species of purple phototrophic bacterium. In these species, solar energy is captured by reaction centre-light harvesting 1 (RC-LH1) complexes which reside in membranes within the cells. The RC comprises three protein subunits: H, M, L and is encircled by a ring of LH1-α and LH1-β subunit pairs. Approximately 10% of these complexes in the wild-type (WT) strain include an additional subunit called protein-W. In this project, we have determined cryo-EM structures for RC-LH1 complexes both with and without protein-W. To enable the purification of RC-LH1-W with 100% occupancy of protein-W, strain expressing a C-terminally His-tagged W was engineered. For complexes lacking W, a knockout strain was used. Proteomic analysis was employed to validate these strains and establish that the WT and W-His expressed similar levels of W relative to RC-LH1.
Project description:To address the question of how photosynthetic bacterium Rhodopseudomonas palustris metabolize lignin derived compound p-coumarate, transcriptomics and quantitative proteomics were combined to characterize gene expression profiles at both the mRNA level and protein level in Rhodopseudomonas palustris grown with succinate, benzoate, and p-coumarate as the carbon source. Keywords: Comparison of transcriptome profiles
Project description:To address the question of how photosynthetic bacterium Rhodopseudomonas palustris metabolize lignin derived compound p-coumarate, transcriptomics and quantitative proteomics were combined to characterize gene expression profiles at both the mRNA level and protein level in Rhodopseudomonas palustris grown with succinate, benzoate, and p-coumarate as the carbon source. Transcriptome profiles among Rhodopseudomonas palustris cells grown with succinate, benzoate, and p-coumarate as the carbon source were compared.
Project description:Rhodopseudomonas palustris strain SA008.1.07 can use syringic acid as sole organic carbon source anaerobically. Grew all anaerobically in various carbon sources: syringic acid, succinate, and p-hydroxybenzoic acid.
Project description:Characterization of post-translational modification of nitrogenase in Rhodopseudomonas palustris strains that produce hydrogen gas constitutively.
