Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Aug;177(16):4801–4804. doi: 10.1128/jb.177.16.4801-4804.1995

Identification of Pseudomonas aeruginosa glpM, whose gene product is required for efficient alginate biosynthesis from various carbon sources.

H P Schweizer 1, C Po 1, M K Bacic 1
PMCID: PMC177247  PMID: 7642508

Abstract

In a mucB (algN) genetic background, insertion of an omega element approximately 200 bp downstream of glpD, encoding sn-glycerol-3-phosphate dehydrogenase from Pseudomonas aeruginosa, had an adverse effect on alginate biosynthesis from various carbon sources. The insertion inactivated glpM, a gene encoding a 12,040-M(r) hydrophobic protein containing 109 amino acids. This protein, which was expressed in a T7 RNA polymerase expression system, appears to be a cytoplasmic membrane protein.

Full Text

The Full Text of this article is available as a PDF (290.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Banerjee P. C., Vanags R. I., Chakrabarty A. M., Maitra P. K. Alginic acid synthesis in Pseudomonas aeruginosa mutants defective in carbohydrate metabolism. J Bacteriol. 1983 Jul;155(1):238–245. doi: 10.1128/jb.155.1.238-245.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Banerjee P. C., Vanags R. I., Chakrabarty A. M., Maitra P. K. Fructose 1,6-bisphosphate aldolase activity is essential for synthesis of alginate from glucose by Pseudomonas aeruginosa. J Bacteriol. 1985 Jan;161(1):458–460. doi: 10.1128/jb.161.1.458-460.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Cuskey S. M., Phibbs P. V., Jr Chromosomal mapping of mutations affecting glycerol and glucose catabolism in Pseudomonas aeruginosa PAO. J Bacteriol. 1985 Jun;162(3):872–880. doi: 10.1128/jb.162.3.872-880.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fellay R., Frey J., Krisch H. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. Gene. 1987;52(2-3):147–154. doi: 10.1016/0378-1119(87)90041-2. [DOI] [PubMed] [Google Scholar]
  6. Franklin M. J., Ohman D. E. Identification of algF in the alginate biosynthetic gene cluster of Pseudomonas aeruginosa which is required for alginate acetylation. J Bacteriol. 1993 Aug;175(16):5057–5065. doi: 10.1128/jb.175.16.5057-5065.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Grinius L., Dreguniene G., Goldberg E. B., Liao C. H., Projan S. J. A staphylococcal multidrug resistance gene product is a member of a new protein family. Plasmid. 1992 Mar;27(2):119–129. doi: 10.1016/0147-619x(92)90012-y. [DOI] [PubMed] [Google Scholar]
  8. Ito K., Sato T., Yura T. Synthesis and assembly of the membrane proteins in E. coli. Cell. 1977 Jul;11(3):551–559. doi: 10.1016/0092-8674(77)90073-3. [DOI] [PubMed] [Google Scholar]
  9. Knutson C. A., Jeanes A. A new modification of the carbazole analysis: application to heteropolysaccharides. Anal Biochem. 1968 Sep;24(3):470–481. doi: 10.1016/0003-2697(68)90154-1. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lewis M. K., Thompson D. V. Efficient site directed in vitro mutagenesis using ampicillin selection. Nucleic Acids Res. 1990 Jun 25;18(12):3439–3443. doi: 10.1093/nar/18.12.3439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ma D., Cook D. N., Hearst J. E., Nikaido H. Efflux pumps and drug resistance in gram-negative bacteria. Trends Microbiol. 1994 Dec;2(12):489–493. doi: 10.1016/0966-842x(94)90654-8. [DOI] [PubMed] [Google Scholar]
  13. Martin D. W., Schurr M. J., Mudd M. H., Deretic V. Differentiation of Pseudomonas aeruginosa into the alginate-producing form: inactivation of mucB causes conversion to mucoidy. Mol Microbiol. 1993 Aug;9(3):497–506. doi: 10.1111/j.1365-2958.1993.tb01711.x. [DOI] [PubMed] [Google Scholar]
  14. May T. B., Chakrabarty A. M. Pseudomonas aeruginosa: genes and enzymes of alginate synthesis. Trends Microbiol. 1994 May;2(5):151–157. doi: 10.1016/0966-842x(94)90664-5. [DOI] [PubMed] [Google Scholar]
  15. May T. B., Shinabarger D., Maharaj R., Kato J., Chu L., DeVault J. D., Roychoudhury S., Zielinski N. A., Berry A., Rothmel R. K. Alginate synthesis by Pseudomonas aeruginosa: a key pathogenic factor in chronic pulmonary infections of cystic fibrosis patients. Clin Microbiol Rev. 1991 Apr;4(2):191–206. doi: 10.1128/cmr.4.2.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ohman D. E., Chakrabarty A. M. Genetic mapping of chromosomal determinants for the production of the exopolysaccharide alginate in a Pseudomonas aeruginosa cystic fibrosis isolate. Infect Immun. 1981 Jul;33(1):142–148. doi: 10.1128/iai.33.1.142-148.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  18. Pavelka M. S., Jr, Hayes S. F., Silver R. P. Characterization of KpsT, the ATP-binding component of the ABC-transporter involved with the export of capsular polysialic acid in Escherichia coli K1. J Biol Chem. 1994 Aug 5;269(31):20149–20158. [PubMed] [Google Scholar]
  19. Schweizer H. P. Allelic exchange in Pseudomonas aeruginosa using novel ColE1-type vectors and a family of cassettes containing a portable oriT and the counter-selectable Bacillus subtilis sacB marker. Mol Microbiol. 1992 May;6(9):1195–1204. doi: 10.1111/j.1365-2958.1992.tb01558.x. [DOI] [PubMed] [Google Scholar]
  20. Schweizer H. P. Improved broad-host-range lac-based plasmid vectors for the isolation and characterization of protein fusions in Pseudomonas aeruginosa. Gene. 1991 Jul 15;103(1):87–92. doi: 10.1016/0378-1119(91)90396-s. [DOI] [PubMed] [Google Scholar]
  21. Schweizer H. P., Po C. Cloning and nucleotide sequence of the glpD gene encoding sn-glycerol-3-phosphate dehydrogenase of Pseudomonas aeruginosa. J Bacteriol. 1994 Apr;176(8):2184–2193. doi: 10.1128/jb.176.8.2184-2193.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schweizer H. P. The agmR gene, an environmentally responsive gene, complements defective glpR, which encodes the putative activator for glycerol metabolism in Pseudomonas aeruginosa. J Bacteriol. 1991 Nov;173(21):6798–6806. doi: 10.1128/jb.173.21.6798-6806.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tisa L. S., Rosen B. P. Molecular characterization of an anion pump. The ArsB protein is the membrane anchor for the ArsA protein. J Biol Chem. 1990 Jan 5;265(1):190–194. [PubMed] [Google Scholar]
  25. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES