Archives of Microbiology

, Volume 162, Issue 5, pp 323–328 | Cite as

Genetic characterization of ompH mutants in the deep-sea bacterium Photobacterium sp. strain SS9

Original Paper


OmpH is an outer membrane protein produced by the deep-sea bacterium Photobacterium species strain SS9 in response to elevated hydrostatic pressure. In order to facilitate studies of the function of this protein, a series of OmpH+ and OmpH- strains were obtained from SS9 by Tn5 gene replacement mutagenesis. A previously isolated ompH::lacZ strain and a derivative of this strain harboring a plasmid expressing the wild-type ompH gene were also utilized. The acridine mutagen ICR 191 preferentially inhibited the growth of OmpH+ over OmpH- cells. Indeed, OmpH+ cultures treated with the mutagen rapidly accumulated mutants producing reduced levels of OmpH. In addition. OmpH+ cells took up the peptide Met-Leu-Phe approximately 15 times more rapidly than OmpH- cells. The results are consistent with the hypothesis that OmpH functions as a relatively large, nonspecific diffusion channel.

Key words

Outer membrane protein Porin Extremophile Piezophile Barophile High pressure adaption Deep-sea bacterium Photobacterium Gene replacement transposon 



Outer membrane protein


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  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410CrossRefPubMedGoogle Scholar
  2. Barlow AK, Heckels JE, Clarke IN (1989) The class 1 outer membrane protein of Neisseria meningitidis: gene sequence and structural and immunological similarities to gonococcal porins. Mol Microbiol 3:131–139Google Scholar
  3. Bartlett D, Wright M, Yayanos A, Silverman M (1989) Isolation of a gene regulated by hydrostatic pressure. Nature 342:572–574Google Scholar
  4. Bartlett DH, Chi E, Wright ME (1993) Sequence of the ompH gene from the deep-sea bacterium Photobacterium SS9. Gene 131:125–128Google Scholar
  5. Benner R, Pakulski JD, McCarthy M, Hedges JI, Hatcher PG (1992) Bulk chemical characteristics of dissolved organic matter in the ocean. Science 255:1561–1564Google Scholar
  6. Better M, Helinski DR (1983) Isolation and characterization of the recA gene of Rhizobium meliloti. J Bacteriol 155:311–316Google Scholar
  7. Braganza LF, Worcester DL (1986) Structural changes in lipid bilayers and biological membranes caused by hydrostatic pressure. Biochemistry 25:7484–7488Google Scholar
  8. Chi E, Bartlett DH (1993) Use of a reporter gene to follow high pressure signal transduction in the deep-sea bacterium Photobacterium SS9. J Bacteriol 175:7533–7540Google Scholar
  9. Cho BC, Azam F (1988) Major role of bacteria in biogeochemical fluxes in the ocean's interior. Nature 332:441–443Google Scholar
  10. DeLong EF (1986) Adaptations of deep-sea bacteria to the abyssal environment. PhD thesis. University of California, San DiegoGoogle Scholar
  11. Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395Google Scholar
  12. Druffel ERM, Williams PM, Bauer JE, Ertel JR (1992) Cycling of dissolved and particulate organic matter in the open ocean. J Geophys Res 97:15,639–15,659Google Scholar
  13. Ducklow HW (1993) Bacterioplankton distributions and production in the northwestern Indian Ocean and Gulf of Oman. Deep-sea Res 40:753–771Google Scholar
  14. Gribskov M, Osguthorpe DJ, Robson B (1978) Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol 120:97–120Google Scholar
  15. Hantke K (1976) Phage T6 — colicin K receptor and nucleoside transport in Escherichia coli. FEBS Lett 70:109–112Google Scholar
  16. Huang H, Hancock REW (1993) Genetic definition of the substrate selectivity of outer membrane porin protein OprD of Pseudomonas aeruginosa. J Bacteriol 175:7793–7800Google Scholar
  17. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedGoogle Scholar
  18. Matsuyama S-I, Inokuchi K, Mizushima S (1984) Promoter exchange between ompF and ompC, genes for osmoregulated major outer membrane proteins of Escherichia coli K-12. J Bacteriol 158:1041–1047Google Scholar
  19. Miller JH (1992) A short course in bacterial genetics. Cold Spring Harbor Press, Cold Spring Harbor, NYGoogle Scholar
  20. Nikaido H (1992) Porins and specific channels of bacterial outer membranes. Mol Microbiol 6:435–442PubMedGoogle Scholar
  21. Nikaido H, Vaara M (1987) Outer membrane. In: Neidhardt FC, Ingraham JL, Low KB, Magasanik B, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella typhimurium cellular and molecular biology, vol 1. American Society for Microbiology, Washington DC, pp 7–22Google Scholar
  22. Poole K, Hancock REW (1986) Isolation of a Tn 501 insertion mutant lacking porin protein P of Pseudomonas aeruginosa. Mol Gen Genet 202:403–409Google Scholar
  23. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  24. Schwartz RM, Dayhoff MO (1978) Matrices for detecting distant relationships. In: Dayhoff MO (ed) Atlas of protein sequences and structure, vol 5. National Biomedical Research Foundation, Washington, DC, pp 353–358Google Scholar
  25. Smith DW (1988) A complete yet flexible system for DNA/protein sequence analysis using VAX/VMS computers. Comput Appl Biosci 4:212–220Google Scholar
  26. Somero GN (1992) Biochemical ecology of deep-sea animals. Experientia 48:537–543Google Scholar
  27. Tabor PS, Ohwada K, Colwell R (1981) Filterable marine bacteria found in the deep sea: distribution, taxonomy, and resposne to starvation. Microb Ecol 7:67–83Google Scholar
  28. Tommassen J, Vermeij P, Struyvé M, Benz R, Poolman JT (1990) Isolation of Neisseria meningitidis mutants deficient in class 1 (PorA) and class 3 (PorB) outer membrane proteins. Infect Immun 58:1355–1359Google Scholar
  29. Trias J, Nikaido H (1990) Protein D2 channel of the Pseudomonas aeruginosa outer membrane has a binding site for basic amino acids and peptides. J Biol Chem 265:15680–15684Google Scholar
  30. Wandersman C, Schwartz M, Ferenci T (1979) Escherichia coli mutants impaired in maltodextrin transport. J Bacteriol 140: 1–13Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  1. 1.Center for Marine Biomedicine and Biotechnology, Scripps Institution of OceanographyUniversity of CaliforniaLa JollaUSA

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