Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Aug;169(8):3785–3791. doi: 10.1128/jb.169.8.3785-3791.1987

Translocation of Vibrio harveyi N,N'-diacetylchitobiase to the outer membrane of Escherichia coli.

M Jannatipour, R W Soto-Gil, L C Childers, J W Zyskind
PMCID: PMC212467  PMID: 3301816

Abstract

The gene encoding N,N'-diacetylchitobiase (chitobiase) of the chitinolytic marine bacterium Vibrio harveyi has been isolated. While expression of the chitobiase gene (chb) was inducible by N,N'-diacetylchitobiose in V. harveyi, it was expressed constitutively when cloned in Escherichia coli, suggesting that controlling elements are not closely linked to chb. Chitobiase was found in the membrane fraction of E. coli cells containing plasmids with the cloned V. harveyi chb gene. When membranes of such cells were separated on Osborn gradients, chitobiase activity was found mainly in the outer membrane band. Translocation of the enzyme to the outer membrane was accompanied by cleavage of a signal peptide. A fusion protein, in which 22 amino acids from the amino terminus of prechitobiase were replaced with 21 amino acids from the pUC19 lacZ amino terminus, was not processed, and 99% of the activity was located in the cytoplasmic fraction. A homology to six amino acids surrounding the lipoprotein processing and modification site was found near the amino terminus of prechitobiase.

Full text

PDF

Images in this article

3789Image
on p.3789

Selected References

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

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Braun V. Covalent lipoprotein from the outer membrane of Escherichia coli. Biochim Biophys Acta. 1975 Oct 31;415(3):335–377. doi: 10.1016/0304-4157(75)90013-1. [DOI] [PubMed] [Google Scholar]
  3. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fuchs R. L., McPherson S. A., Drahos D. J. Cloning of a Serratia marcescens Gene Encoding Chitinase. Appl Environ Microbiol. 1986 Mar;51(3):504–509. doi: 10.1128/aem.51.3.504-509.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fujimura F. K., Zyskind J. W., Smith D. W. The Escherichia coli dnaB protein is required for initiation of chromosomal DNA replication. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):559–562. doi: 10.1101/sqb.1979.043.01.061. [DOI] [PubMed] [Google Scholar]
  6. GAREN A., LEVINTHAL C. A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase. Biochim Biophys Acta. 1960 Mar 11;38:470–483. doi: 10.1016/0006-3002(60)91282-8. [DOI] [PubMed] [Google Scholar]
  7. Jones J. D., Grady K. L., Suslow T. V., Bedbrook J. R. Isolation and characterization of genes encoding two chitinase enzymes from Serratia marcescens. EMBO J. 1986 Mar;5(3):467–473. doi: 10.1002/j.1460-2075.1986.tb04235.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kahn M., Kolter R., Thomas C., Figurski D., Meyer R., Remaut E., Helinski D. R. Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol. 1979;68:268–280. doi: 10.1016/0076-6879(79)68019-9. [DOI] [PubMed] [Google Scholar]
  9. Koshland D., Botstein D. Secretion of beta-lactamase requires the carboxy end of the protein. Cell. 1980 Jul;20(3):749–760. doi: 10.1016/0092-8674(80)90321-9. [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. Matsumura P., Silverman M., Simon M. Synthesis of mot and che gene products of Escherichia coli programmed by hybrid ColE1 plasmids in minicells. J Bacteriol. 1977 Dec;132(3):996–1002. doi: 10.1128/jb.132.3.996-1002.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Monreal J., Reese E. T. The chitinase of Serratia marcescens. Can J Microbiol. 1969 Jul;15(7):689–696. doi: 10.1139/m69-122. [DOI] [PubMed] [Google Scholar]
  13. Nakamura K., Inouye M. DNA sequence of the gene for the outer membrane lipoprotein of E. coli: an extremely AT-rich promoter. Cell. 1979 Dec;18(4):1109–1117. doi: 10.1016/0092-8674(79)90224-1. [DOI] [PubMed] [Google Scholar]
  14. Nealson K. H., Hastings J. W. Bacterial bioluminescence: its control and ecological significance. Microbiol Rev. 1979 Dec;43(4):496–518. doi: 10.1128/mr.43.4.496-518.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. REISSIG J. L., STORMINGER J. L., LELOIR L. F. A modified colorimetric method for the estimation of N-acetylamino sugars. J Biol Chem. 1955 Dec;217(2):959–966. [PubMed] [Google Scholar]
  17. Reichelt J. L., Baumann P., Baumann L. Study of genetic relationships among marine species of the genera Beneckea and Photobacterium by means of in vitro DNA/DNA hybridization. Arch Microbiol. 1976 Oct 11;110(1):101–120. doi: 10.1007/BF00416975. [DOI] [PubMed] [Google Scholar]
  18. Wortman A. T., Somerville C. C., Colwell R. R. Chitinase determinants of Vibrio vulnificus: gene cloning and applications of a chitinase probe. Appl Environ Microbiol. 1986 Jul;52(1):142–145. doi: 10.1128/aem.52.1.142-145.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Zyskind J. W., Cleary J. M., Brusilow W. S., Harding N. E., Smith D. W. Chromosomal replication origin from the marine bacterium Vibrio harveyi functions in Escherichia coli: oriC consensus sequence. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1164–1168. doi: 10.1073/pnas.80.5.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES