Isolation and partial characterization of Rhodopseudomonas sphaeroides mutants defective in the regulation of ribulose bisphosphate carboxylase/oxygenase

Weaver, Keith E.; Tabita, F. Robert

doi:10.1128/jb.156.2.507-515.1983

Cited by 60 publications

(14 citation statements)

References 18 publications

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“…The antibiotic concentrations used for selection of transconjugants and routine plasmid maintenance have been described previously (3). Photosynthetic growth on solid medium was achieved as described previously; 1.8% Bacto-Agar (Difco Laboratories, Detroit, Mich.) was added to minimal medium, and the plates were incubated in anaerobic jars in a C02-H2 atmosphere in the light (20).…”

Section: Methodsmentioning

confidence: 99%

Expression and regulation of Bradyrhizobium japonicum and Xanthobacter flavus CO2 fixation genes in a photosynthetic bacterial host

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Tabita

1993

J Bacteriol

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Calvin cycle carbon dioxide fixation genes encoded on DNA fragments from two nonphotosynthetic, chemolithoautotrophic bacteria, Bradyrhizobium japonicum and Xanthobacter flavus, were found to complement and support photosynthetic growth of a ribulose 1,5-bisphosphate carboxylase-oxygenase (RubisCO) deletion mutant of the purple nonsulfur bacterium Rhodobacter sphaeroides. The regulation of RubisCO expression was analyzed in the complemented R. sphaeroides RubisCO deletion mutant. Distinct differences in the regulation of RubisCO synthesis were revealed when the complemented R. sphaeroides strains were cultured under photolithoautotrophic and photoheterotrophic growth conditions, e.g., a reversal in the normal pattern of RubisCO gene expression. These studies suggest that sequences and molecular signals which regulate the expression of diverse RubisCO genes may be probed by using the R. sphaeroides complementation system.

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Section: Methodsmentioning

confidence: 99%

Expression and regulation of Bradyrhizobium japonicum and Xanthobacter flavus CO2 fixation genes in a photosynthetic bacterial host

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Tabita

1993

J Bacteriol

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“…Mutants of Rb. sphaeroides have been isolated which are unable to grow photoautotrophically or on butyrate, and are unable to increase the levels of form I Rubisco under CO2-1imitation, although the level of form II enzyme was similar to that of the wild-type [85]. These results suggested that the mutants were regulatory mutants, and that synthesis of the two forms of the enzyme is regulated independently.…”

Section: Co 2 Fixation During Phototrophic Growthmentioning

confidence: 97%

“…The level of form I enzyme increases greatly during photoautotrophic growth, or during growth on a highly reduced substrate such as butyrate, both in Rb. sphaeroides [85] and in Rb. capsulatus [86].…”

Section: Co 2 Fixation During Phototrophic Growthmentioning

confidence: 99%

Biochemical genetics revisited: the use of mutants to study carbon and nitrogen metabolism in the photosynthetic bacteria

Willison¹

1993

FEMS Microbiology Letters

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The biochemical genetics approach is defined as the use of mutants, in comparative studies with the wild‐type, to obtain information about biochemical and physiological processes in complex metabolic systems. This approach has been used extensively, for example in studies on the bioenergetics of the photosynthetic bacteria, but has been applied less frequently to studies of intermediary carbon and nitrogen metabolism in phototrophic organisms. Several important processes in photosynthetic bacteria—the regulation of nitrogenase synthesis and activity, the control of intracellular redox balance during photoheterotrophic growth, and chemotaxis—have been shown to involve metabolism. However, current understanding of carbon and nitrogen metabolism in these organisms is insufficient to allow a complete understanding of these phenomena. The purpose of the present review is to give an overview of carbon and nitrogen metabolism in the photosynthetic bacteria, with particular emphasis on work carried out with mutants, and to indicate areas in which the biochemical genetics approach could be applied successfully. In particular, it will be argued that, in the case of Rhodobacter capsulatus and Rb. sphaeroides, two species which are fast‐growing, possess a versatile metabolism, and have been extensively studied genetically, it should be possible to obtain a complete, integrated description of carbon and nitrogen metabolism, and to undertake a qualitative and quantitative analysis of the flow of carbon and reducing equivalents during photoheterotrophic growth. This would require a systematic biochemical genetic study employing techniques such as HPLC, NMR, and mass spectrometry, which are briefly discussed. The review is concerned mainly with Rb. capsulatus and Rb. sphaeroides, since most studies with mutants have been carried out with these organisms. However, where possible, a comparison is made with other species of purple non‐sulphur bacteria and with purple and green sulphur bacteria, and recent literature relevant to these organisms has been cited.

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“…Escherichia coli JM107 (60), JM109 pir, and S17-1 pir (40) were grown aerobically on LB medium (2) at 37°C. Aerobic cultures of R. capsulatus were grown in PYE medium (57) at 30°C. Photosynthetic cultures were grown in Ormerod's medium (37) supplemented with thiamine (1 g/ml), nicotinic acid (1 g/ml), and biotin (0.1 g/ml).…”

Section: Methodsmentioning

confidence: 99%

“…For gene disruption experiments, plasmid pJP5603 derivatives were conjugated into R. capsulatus SB1003 by using E. coli S17-1 pir (40). For complementation of mutant strains, plasmids were conjugated into R. capsulatus by triparental matings on filter pads as previously described (57), using the helper plasmid pRK2013 (10). Southern blotting and hybridization.…”

Section: Methodsmentioning

confidence: 99%

Physiological Control and Regulation of the Rhodobacter capsulatus cbb Operons

1998

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The genes encoding enzymes of the Calvin-Benson-Bassham (CBB) reductive pentose phosphate pathway in Rhodobacter capsulatus are organized in at least two operons, each preceded by a separate cbbR gene, encoding potential LysR-type transcriptional activators. As a prelude to studies ofcbb gene regulation in R. capsulatus, the nucleotide sequence of a 4,537-bp region, which includedcbbR II, was determined. This region contained the following open reading frames: a partial pgmgene (encoding phosphoglucomutase) and a complete qorgene (encoding NADPH:quinone oxidoreductase), followed by cbbR II, cbbF (encoding fructose 1,6-bisphosphatase), cbbP (encoding phosphoribulokinase), and part of cbbT (encoding transketolase). Physiological control of the CBB pathway and regulation of the R. capsulatus cbb genes were studied by using a combination of mutant strains and promoter fusion constructs. Characterization of mutant strains revealed that either form I or form II ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO), encoded by the cbbLS andcbbM genes, respectively, could support photoheterotrophic and autotrophic growth. A strain with disruptions in both cbbL and cbbM could not grow autotrophically and grew photoheterotrophically only when dimethyl sulfoxide was added to the culture medium. Disruption ofcbbP resulted in a strain that did not synthesize form II RubisCO and had a phenotype similar to that observed in the RubisCO-minus strain, suggesting that there is only onecbbP gene in R. capsulatus and that this gene is cotranscribed with cbbM. Analysis of RubisCO activity and synthesis in strains with disruptions in eithercbbR I orcbbR II, and β-galactosidase determinations from wild-type and mutant strains containing cbb Ip- andcbb IIp-lacZ fusion constructs, indicated that the cbb I andcbb II operons of R. capsulatus are within separate CbbR regulons.

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Isolation and partial characterization of Rhodopseudomonas sphaeroides mutants defective in the regulation of ribulose bisphosphate carboxylase/oxygenase

Cited by 60 publications

References 18 publications

Expression and regulation of Bradyrhizobium japonicum and Xanthobacter flavus CO2 fixation genes in a photosynthetic bacterial host

Expression and regulation of Bradyrhizobium japonicum and Xanthobacter flavus CO2 fixation genes in a photosynthetic bacterial host

Biochemical genetics revisited: the use of mutants to study carbon and nitrogen metabolism in the photosynthetic bacteria

Physiological Control and Regulation of the Rhodobacter capsulatus cbb Operons

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