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Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Feb;149(2):576–586. doi: 10.1128/jb.149.2.576-586.1982

Characterization of factor IIIGLc in catabolite repression-resistant (crr) mutants of Salmonella typhimurium.

B J Scholte, A R Schuitema, P W Postma
PMCID: PMC216545  PMID: 7035434

Abstract

crr mutants of Salmonella typhimurium are thought to be defective in the regulation of adenylate cyclase and a number of transport systems by the phosphoenolpyruvate-dependent sugar phosphotransferase system, crr mutants are also defective in the enzymatic activity of factor IIIGlc (IIIGlc), a protein component of the phosphotransferase system involved in glucose transport. Therefore, it has been proposed that IIIGlc is the primary effector of phosphotransferase system-mediated regulation of cell metabolism. We characterized crr mutants with respect to the presence and function of IIIGlc by using an immunochemical approach. All of the crr mutants tested had low (0 to 30%) levels of IIIGlc compared with wild-type cells, as determined by rocket immunoelectrophoresis. The IIIGlc isolated from one crr mutant was investigated in more detail and showed abnormal aggregation behavior, which indicated a structural change in the protein. These results supported the hypothesis that a crr mutation directly affects IIIGlc, probably by altering the structural gene of IIIGlc. Several crr strains which appeared to be devoid of IIIGlc in immunoprecipitation assays were still capable of in vitro phosphorylation and transport of methyl alpha-glucoside. This phosphorylation activity was sensitive to specific anti-IIIGlc serum. Moreover, the membranes of crr mutants, as well as those of wild-type cells, contained a protein that reacted strongly with our anti-IIIGlc serum. We propose that S. typhimurium contains a membrane-bound form of IIIGlc which may be involved in phosphotransferase system activity.

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Selected References

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  1. Boos W., Lengeler J., Hermann K. O., Unsöld H. J. The regulation of the beta-methylgalactoside transport system and of the galactose binding protein of Escherichia coli K12. Eur J Biochem. 1971 Apr 30;19(4):457–470. doi: 10.1111/j.1432-1033.1971.tb01336.x. [DOI] [PubMed] [Google Scholar]
  2. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  3. Cordaro J. C., Roseman S. Deletion mapping of the genes coding for HPr and enzyme I of the phosphoenolpyruvate: sugar phosphotransferase system in Salmonella typhimurium. J Bacteriol. 1972 Oct;112(1):17–29. doi: 10.1128/jb.112.1.17-29.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dills S. S., Apperson A., Schmidt M. R., Saier M. H., Jr Carbohydrate transport in bacteria. Microbiol Rev. 1980 Sep;44(3):385–418. doi: 10.1128/mr.44.3.385-418.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feucht B. U., Saier M. H., Jr Fine control of adenylate cyclase by the phosphoenolpyruvate:sugar phosphotransferase systems in Escherichia coli and Salmonella typhimurium. J Bacteriol. 1980 Feb;141(2):603–610. doi: 10.1128/jb.141.2.603-610.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gottesman S., Zipser D. Deg phenotype of Escherichia coli lon mutants. J Bacteriol. 1978 Feb;133(2):844–851. doi: 10.1128/jb.133.2.844-851.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Harwood J. P., Peterkofsky A. Glucose-sensitive adenylate cyclase in toluene-treated cells of Escherichia coli B. J Biol Chem. 1975 Jun 25;250(12):4656–4662. [PubMed] [Google Scholar]
  8. Hays J. B., Simoni R. D., Roseman S. Sugar transport. V. A trimeric lactose-specific phosphocarrier protein of the Staphylococcus aureus phosphotransferase system. J Biol Chem. 1973 Feb 10;248(3):941–956. [PubMed] [Google Scholar]
  9. Jones-Mortimer M. C., Kornberg H. L., Maltby R., Watts P. D. Role of the crr-gene in glucose uptake by Escherichia coli. FEBS Lett. 1977 Feb 15;74(1):17–19. doi: 10.1016/0014-5793(77)80742-4. [DOI] [PubMed] [Google Scholar]
  10. Kemper J. Gene order and co-transduction in the leu-ara-fol-pyrA region of the Salmonella typhimurium linkage map. J Bacteriol. 1974 Jan;117(1):94–99. doi: 10.1128/jb.117.1.94-99.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kornberg H. L., Watts P. D. tgs and crr: Genes involved in catabolite inhibition and inducer exclusion in Escherichia coli. FEBS Lett. 1979 Aug 15;104(2):313–316. doi: 10.1016/0014-5793(79)80841-8. [DOI] [PubMed] [Google Scholar]
  12. Kornberg H., Watts P. D., Brown K. Mechanisms of 'inducer exclusion' by glucose. FEBS Lett. 1980 Aug 25;117 (Suppl):K28–K36. doi: 10.1016/0014-5793(80)80567-9. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Mount D. W. The genetics of protein degradation in bacteria. Annu Rev Genet. 1980;14:279–319. doi: 10.1146/annurev.ge.14.120180.001431. [DOI] [PubMed] [Google Scholar]
  15. Overbeeke N., Van Scharrenburg G., Lugtenberg B. Antigenic relationships between pore proteins of Escherichia coli K12. Eur J Biochem. 1980 Sep;110(1):247–254. doi: 10.1111/j.1432-1033.1980.tb04862.x. [DOI] [PubMed] [Google Scholar]
  16. Peterkofsky A., Gazdar C. Glucose inhibition of adenylate cyclase in intact cells of Escherichia coli B. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2324–2328. doi: 10.1073/pnas.71.6.2324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Postma P. W. Defective enzyme II-BGlc of the phosphoenolpyruvate:sugar phosphotransferase system leading to uncoupling of transport and phosphorylation in Salmonella typhimurium. J Bacteriol. 1981 Aug;147(2):382–389. doi: 10.1128/jb.147.2.382-389.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Postma P. W., Roseman S. The bacterial phosphoenolpyruvate: sugar phosphotransferase system. Biochim Biophys Acta. 1976 Dec 14;457(3-4):213–257. doi: 10.1016/0304-4157(76)90001-0. [DOI] [PubMed] [Google Scholar]
  19. Postma P. W., Schuitema A., Kwa C. Regulation of methyl beta-galactoside permease activity in pts and crr mutants of Salmonella typhimurium. Mol Gen Genet. 1981;181(4):448–453. doi: 10.1007/BF00428734. [DOI] [PubMed] [Google Scholar]
  20. Saier M. H., Jr, Feucht B. U. Coordinate regulation of adenylate cyclase and carbohydrate permeases by the phosphoenolpyruvate:sugar phosphotransferase system in Salmonella typhimurium. J Biol Chem. 1975 Sep 10;250(17):7078–7080. [PubMed] [Google Scholar]
  21. Saier M. H., Jr, Feucht B. U. Regulation of carbohydrate transport activities in Salmonella typhimurium: use of the phosphoglycerate transport system to energize solute uptake. J Bacteriol. 1980 Feb;141(2):611–617. doi: 10.1128/jb.141.2.611-617.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Saier M. H., Jr, Roseman S. Sugar transport. 2nducer exclusion and regulation of the melibiose, maltose, glycerol, and lactose transport systems by the phosphoenolpyruvate:sugar phosphotransferase system. J Biol Chem. 1976 Nov 10;251(21):6606–6615. [PubMed] [Google Scholar]
  23. Saier M. H., Jr, Roseman S. Sugar transport. The crr mutation: its effect on repression of enzyme synthesis. J Biol Chem. 1976 Nov 10;251(21):6598–6605. [PubMed] [Google Scholar]
  24. Saier M. H., Jr, Straud H., Massman L. S., Judice J. J., Newman M. J., Feucht B. U. Permease-specific mutations in Salmonella typhimurium and Escherichia coli that release the glycerol, maltose, melibiose, and lactose transport systems from regulation by the phosphoenolpyruvate:sugar phosphotransferase system. J Bacteriol. 1978 Mar;133(3):1358–1367. doi: 10.1128/jb.133.3.1358-1367.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Scholte B. J., Postma P. W. Competition between two pathways for sugar uptake by the phosphoenolpyruvate-dependent sugar phosphotransferase system in Salmonella typhimurium. Eur J Biochem. 1981;114(1):51–58. doi: 10.1111/j.1432-1033.1981.tb06171.x. [DOI] [PubMed] [Google Scholar]
  26. Scholte B. J., Postma P. W. Mutation in the crp gene of Salmonella typhimurium which interferes with inducer exclusion. J Bacteriol. 1980 Feb;141(2):751–757. doi: 10.1128/jb.141.2.751-757.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Scholte B. J., Schuitema A. R., Postma P. W. Isolation of IIIGlc of the phosphoenolpyruvate-dependent glucose phosphotransferase system of Salmonella typhimurium. J Bacteriol. 1981 Oct;148(1):257–264. doi: 10.1128/jb.148.1.257-264.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Waygood E. B., Meadow N. D., Roseman S. Modified assay procedures for the phosphotransferase system in enteric bacteria. Anal Biochem. 1979 May;95(1):293–304. doi: 10.1016/0003-2697(79)90219-7. [DOI] [PubMed] [Google Scholar]

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