Reversible regulation of the nitrogenase iron protein from Rhodospirillum rubrum by ADP-ribosylation in vitro

Lowery, Robert G.; Saari, Leonard L.; Ludden, Paul W.

doi:10.1128/jb.166.2.513-518.1986

Cited by 96 publications

(76 citation statements)

References 44 publications

(28 reference statements)

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“…This is also supported by results obtained with the form II deletion strain. It is reasonable to speculate that the inactivation-reactivation process somehow depends on the nitrogen status of the cells, reminiscent of the ammonia switch-off, switch-on regulation of nitrogenase activity of purple nonsulfur photosynthetic bacteria (24,42). Certainly, as demonstrated in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Thus, it is essential for CO2 fixation to be regulated. In addition to CO2 fixation, purple nonsulfur photosynthetic bacteria catalyze another energy-costly process, nitrogen fixation, which has been intensively investigated genetically and biochemically (9,23,24,30). At this time, a full understanding of the regulation of microbial carbon dioxide fixation is lacking.…”

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confidence: 99%

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Reversible inactivation and characterization of purified inactivated form I ribulose 1,5-bisphosphate carboxylase/oxygenase of Rhodobacter sphaeroides

Wang

Tabita

1992

J Bacteriol

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(31,40) and, in addition, reversible binding of the intracellular inhibitor 2-carboxy-arabinitol-1-phosphate to carbamylated RubisCO (3,15). In bacteria, there have been periodic indications that RubisCO might be subject to some form of modification or alteration of its activity in vivo (17,21,34), and recent work with the photosynthetic bacterium Rhodomicrobium vannielii suggests a covalent phosphorylation (26); in this latter study, however, no information relative to how phosphorylation affected RubisCO activity was presented. In other photosynthetic bacteria, particularly Rhodobacter sphaeroides and Rhodobacter capsulatus, two different forms of RubisCO are synthesized (12,13,33). The form I enzyme resembles the enzyme that is widely distributed in plants, algae, and most bacteria and contains both large (catalytic) and small subunits (L8S8); the form II RubisCO is composed only of large subunits which show little homology to form I large subunits (11,14,37). At the molecular level, the form I RubisCO genes of R. sphaeroi-* Corresponding author.des, rbcL and rbcS, and the form II RubisCO gene of R. sphaeroides, rbpL, are found in distinct chromosomal operons (10, 11), and there is evidence to support both independent and interdependent regulation of these genes (6,11,16,19). As for the regulation of RubisCO activity in this organism, it was recently found that the form I enzyme is specifically inactivated when metabolizable organic acids are added to cells growing with CO2 as the sole source of carbon (20).The results of the present investigation further characterize the inactivation of the form I RubisCO of R. sphaeroides both in vivo and in vitro and give further credence to the possibility that this enzyme is subject to a reversible modification of its activity in the cell.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Reversible inactivation and characterization of purified inactivated form I ribulose 1,5-bisphosphate carboxylase/oxygenase of Rhodobacter sphaeroides

Wang

Tabita

1992

J Bacteriol

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“…Post-translational regulation of nitrogenase activity is best-characterized in Rhodospirillum rubrum (Pope et al, 1985;Lowery et al, 1986;Saari et al, 1986;Lowery & Ludden, 1988;Liang et al, 1991). Inactivation of nitrogenase reductase by ADP-ribosylation is catalysed by the draT gene product (dinitrogenase reductase ADPribosyltransferase) in response to ammonia, oxygen or darkness.…”

Section: Introductionmentioning

confidence: 99%

The draTG gene region of Rhodobacter capsulatus is required for post-translational regulation of both the molybdenum and the alternative nitrogenase

Masepohl

Krey

Klipp

1993

Journal of General Microbiology

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Synthetic oligonucleotides, which were designed according to amino acid sequences conserved between Rhodospirillum rubrum and Azospirillum brasileizse DraT and DraG, respectively, were used to identify the corresponding genes of Rhodobacter capsulatus. Sequence analysis of a 1904bp DNA fragment proved the existence of R. capsulatus draT and draG. These two genes were separated by 11 bp only, suggesting that R. capsulatus draT and draG were part of one transcriptional unit. In contrast to R. rubrum, A. brasilense and AzospirUum lipoferum, the R. capsulatus draTG genes were not located upstream of the structural genes of nitrogenase nzmDK but close to the dctP gene at a distance of about lo00 kb from the nzjHDK genes. Deletion mutations in the draTG gene region were constructed and introduced into R. capsulatus wild-type and a nijHDK deletion strain. The resulting mutant strains were examined for post-translational regulation of the molybdenum and the alternative nitrogenase in response to ammonia and darkness. Under 'switch-off' conditions the modified (ADP-ribosylated) and the non-modified forms of component I1 of both the molybdenum and the alternative nitrogenase were detected in a ,draTG wild-type background by immunoblot analysis, whereas only the nonmodified forms were present in the draTG deletion strains. Nitrogenase activity in these strains was followed by the acetylene reduction assay. In contrast to the wild-type, draTG mutants were not affected in nitrogenase activity in response to ammonia or darkness. These results demonstrated that the draTG genes are required for posttranslational regulation of both the molybdenum and the heterometal-free nitrogenase in R. capsulatus.

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“…The NAD ϩ -dependent ADP-ribosylation of R101 of the R. rubrum Fe protein is catalyzed by dinitrogenase reductase ADP-ribosyltransferase (DRAT) in response to energy limitation or nitrogen sufficiency (16). Upon relief of these negative stimuli, dinitrogenase reductase-activating glycohydrolase (DRAG) removes the ADP-ribose moiety, restoring the original, fully active Fe protein (17,29).…”

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Effects of Perturbations of the Nitrogenase Electron Transfer Chain on Reversible ADP-Ribosylation of Nitrogenase Fe Protein in Klebsiella pneumoniae Strains Bearing the Rhodospirillum rubrum dra Operon

et al. 2000

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The redox state of nitrogenase Fe protein is shown to affect regulation of ADP-ribosylation in Klebsiella pneumoniae strains transformed by plasmids carrying dra genes from Rhodospirillum rubrum. The dra operon encodes dinitrogenase reductase ADP-ribosyltransferase and dinitrogenase reductase-activating glycohydrolase, enzymes responsible for the reversible inactivation, via ADP-ribosylation, of nitrogenase Fe protein in R. rubrum. In bacteria containing the dra operon in their chromosomes, inactivation occurs in response to energy limitation or nitrogen sufficiency. The dra gene products, expressed at a low level in K. pneumoniae, enable transformants to reversibly ADP-ribosylate nitrogenase Fe protein in response to the presence of fixed nitrogen. The activities of both regulatory enzymes are regulated in vivo as described in R. rubrum. Genetic perturbations of the nitrogenase electron transport chain were found to affect the rate of inactivation of Fe protein. Strains lacking the electron donors to Fe protein (NifF or NifJ) were found to inactivate Fe protein more quickly than a strain with wild-type background. Deletion of nifD, which encodes a subunit of nitrogenase MoFe protein, was found to result in a slower inactivation response. No variation was found in the reactivation responses of these strains. It is concluded that the redox state of the Fe protein contributes to the regulation of the ADP-ribosylation of Fe protein.Biological nitrogen fixation is a capability found in diverse genera of bacteria, including plant root-associated symbionts, free-living bacteria, and cyanobacteria. The conversion of N 2 to NH 3 is catalyzed by the nitrogenase enzyme complex, which comprises two separable components. Nitrogenase MoFe protein is an ␣ 2 ␤ 2 tetramer containing the cofactor FeMoco, believed to be the site of nitrogen reduction. Nitrogenase Fe protein is the obligate electron donor to MoFe protein. Fe protein is a homodimer, containing a [Fe 4 S 4 ] cluster bound by cysteinyl ligands at the dimer interface (reviewed in reference 9). Nitrogen fixation is extremely expensive in terms of the biological energy requirement. No fewer than 16 high-energy phosphoanhydride bond cleavages (ATP energy equivalents) are required for the reduction of a single dinitrogen molecule (21). In response to this requirement in the unstable microbial environment, some nitrogen-fixing bacteria have adopted a posttranslational regulatory system in which the activity of the electron carrier, Fe protein, is regulated. Inhibition of Fe protein activity is achieved by ADP-ribosylation of a specific arginine residue located on the surface of Fe protein that interacts with MoFe protein (6, 26).The ADP-ribosylation system has been best characterized in the purple nonsulfur bacterium Rhodospirillum rubrum. The NAD ϩ -dependent ADP-ribosylation of R101 of the R. rubrum Fe protein is catalyzed by dinitrogenase reductase ADP-ribosyltransferase (DRAT) in response to energy limitation or nitrogen sufficiency (16). Upon relief of these negative ...

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Reversible regulation of the nitrogenase iron protein from Rhodospirillum rubrum by ADP-ribosylation in vitro

Cited by 96 publications

References 44 publications

Reversible inactivation and characterization of purified inactivated form I ribulose 1,5-bisphosphate carboxylase/oxygenase of Rhodobacter sphaeroides

Reversible inactivation and characterization of purified inactivated form I ribulose 1,5-bisphosphate carboxylase/oxygenase of Rhodobacter sphaeroides

The draTG gene region of Rhodobacter capsulatus is required for post-translational regulation of both the molybdenum and the alternative nitrogenase

Effects of Perturbations of the Nitrogenase Electron Transfer Chain on Reversible ADP-Ribosylation of Nitrogenase Fe Protein in Klebsiella pneumoniae Strains Bearing the Rhodospirillum rubrum dra Operon

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