Covalent modification of proteins in Bacillus subtilis during the process of sporulation, germination and outgrowth

Köhler, E.; Antranikian, Garabed

doi:10.1016/0378-1097(89)90152-3

Cited by 4 publications

(4 citation statements)

References 30 publications

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“…The precursors for protein synthesis are believed to be supplied by turnover of the proteins during germination and sporulation. This accounts for the massive degradation that has been observed during such stages (1,4,10,21,23 subtilis were induced to sporulate as described in the legend to Fig. 7.…”

Section: Discussionmentioning

confidence: 68%

Temporal expression of a membrane-associated protein putatively involved in repression of initiation of DNA replication in Bacillus subtilis

et al. 1992

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A Bacillus subtilis membrane-associated protein that binds specifically to the origin region of DNA replication may act as an inhibitor of DNA replication (J. Laffan and W. Firshein, Proc. Natl. Acad. Sci. USA 85:7452-7456, 1988). This protein, originally estimated to be 64 kDa, had a slightly lower molecular size (57 kDa), as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis during these studies. The size difference may be due to processing that results in modification of the protein. The protein can be extracted from both cytosol and membrane fractions, and the amounts in these fractions vary during the developmental cycle of B. subtilis. A complex pattern of expression in which significant levels were detected in spores was revealed; levels decreased dramatically during germination and increased after the first round of DNA replication. The decrease during germination was due to protease activity, as demonstrated by the addition of protease inhibitors and radioactive-labeling chase experiments. During vegetative growth, the protein levels increased until stationary phase, after which there was another decrease during sporulation. The decrease during sporulation may be partially due to sequestering of the protein into forespores, since as the putative repressor protein decreased in the mother cell, it increased in the forespores. However, protease activity was also involved in the decrease in the mother cell. The changes in expression of this protein are consistent with its role as a repressor of initiation of DNA replication. Additional studies, including sequence analysis and further antibody analysis, show that this protein is not a subunit of the pyruvate dehydrogenase complex. This relationship had been a possibility based upon the results of others (H. Hemila, A. Pavla, L. Paulin, S.Arvidson, and I. Palva, J. Bacteriol. 172:5052-5063, 1990

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Section: Discussionmentioning

confidence: 68%

Temporal expression of a membrane-associated protein putatively involved in repression of initiation of DNA replication in Bacillus subtilis

et al. 1992

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“…The focus of this study was whether the stress response in C. acetobutylicum involved regulation by a reversible protein phosphorylation mechanism. It has been clearly shown in a number of bacterial systems, including clostridial species, that growth conditions alter the patterns of phosphorylated proteins (1,7,11,12,15,18,21). However, in E. coli, heat or ethanol stress did not cause changes in the extent of Heat stressing the C. acetobutylicum cultures resulted in increased total 32p labeling of proteins at each growth phase tested, as seen by comparing cell extracts after the temperature upshift (odd-numbered lanes) with those of unstressed cells (even-numbered lanes).…”

Section: Methodsmentioning

confidence: 99%

“…More importantly, considerable differences are seen in the patterns and intensities of 32P-labeled proteins under different growth conditions. For example, the phosphoprotein bands differ in Bradyrhizobium japonicum, in cells growing ex planta and in bacteroids (11), and in Bacillus subtilis, during the sporulation, germination, and outgrowth stages of the growth cycle (12). Variation with growth condition is also seen in the phosphoproteins of the anaerobic sporeformer Clostridium thermohydrosulfuricum (15).…”

mentioning

confidence: 99%

Protein phosphorylation in response to stress in Clostridium acetobutylicum

Balodimos

Rapaport

Kashket

1990

Appl Environ Microbiol

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The possible involvement of protein phosphorylation in the clostridial stress response was investigated by radioactively labeling growing cells of Clostridium acetobutylicum with 32Pi or cell extracts with [y-32P]ATP. Several phosphoproteins were identified; these were not affected by the growth stage of the culture. Although the extent of protein phosphorylation was increased by heat stress, the phosphoproteins did not correspond to known stress proteins seen in one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purified clostridial DnaK, a stress protein, acted as a kinase catalyzing the phosphorylation of a 50-kilodalton protein. The phosphorylation of this protein was enhanced in extracts prepared from heat-stressed cells.

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“…In this system, the activity of one protein, HPr, is also regulated by ATP-dependent phosphorylation of a serine residue (14,16). Changes in protein phosphorylation during sporulation, germination, and outgrowth have also been described for B. subtilis by using in vivo and in vitro assays (9), but nQne of the modified proteins were identified. Previously, we reported changes in phosphloproteins during sporulation of Bacillus megaterium (12a) and B. subtili.…”

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confidence: 99%

Identification of proteins phosphorylated by ATP during sporulation of Bacillus subtilis

1992

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Protein phosphorylation in BaciUus subtilis was assayed in vitro by using extracts prepared from cells at various times during'growth and sporulation. At least six proteins were labeled in vitro by using [_y-32PATP and extracts of ve'getative cells. In extracts prepared at the end of exponential growth and during'stationary phase, 12 to 13 proteins were labeled. Seven of the phosphoproteins were purified' by fast-performance liquid chromatography and polyacrylamide gel electrophoresis, blotted to Immobilon membranes, and subjected to partial protein sequencing. One of the sequences had sequence homology (>45%) to elongation factor G from several bacterial species, and four seqwences matched the predicted amino-terminal sequences of the outB, orJY-tsr, oiffl, and ptsH genes.Protein phosphorylation, an established regulatory mechanism in eukaryotes (4), has been observed for many bacterial species (3, 16). One example of prokaryotic protein phosphorylation occurs in the sensor-kinase and responseregulator systems, in which one of the proteins is an environment-sensing kinase and the other is a phosphorylation substrate and transcriptional regulator. Four pairs of these proteins have been identified in Bacillus subtilis (20); they are involved in genetic competence (ComnP-ComA), phosphorus utilization (PhoR-PhoB), expression of secreted proteases (DegS-DegU), and sporulation (KinA-SpoOA). The latter involves the autophosphorylation of a histidine residue on KinA and subsequent transfer of the phosphate group to aspartate residues of proteins SpoOF, SpoOB, and SpoOA, a transcriptional regulator (2). Another example of protein phosphorylation is the phosphoenolpyruvate:sugar phosphotransferase system for sugar transport, in which the phosphate from phosphoenolpyruvate is' transferred, via specific histidine residues of several proteins (HPr, enzymes I and II/III), to a sugar (16). In this system, the activity of one protein, HPr, is also regulated by ATP-dependent phosphorylation of a serine residue (14,16). Changes in protein phosphorylation during sporulation, germination, and outgrowth have also been described for B. subtilis by using in vivo and in vitro assays (9), but nQne of the modified proteins were identified. Previously, we reported changes in phosphloproteins during sporulation of Bacillus megaterium (12a) and B. subtili. '(12). In this study, we have identified the N-termipal amino acid seqpences of several proteins that wfere phosphorylated in B. subtilis.

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Covalent modification of proteins in Bacillus subtilis during the process of sporulation, germination and outgrowth

Cited by 4 publications

References 30 publications

Temporal expression of a membrane-associated protein putatively involved in repression of initiation of DNA replication in Bacillus subtilis

Temporal expression of a membrane-associated protein putatively involved in repression of initiation of DNA replication in Bacillus subtilis

Protein phosphorylation in response to stress in Clostridium acetobutylicum

Identification of proteins phosphorylated by ATP during sporulation of Bacillus subtilis

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