Abstract
The cellular location of the haemolysin of Vibrio cholerae El Tor strain 017 has been analyzed. This protein is found both in the periplasmic space and the extracellular medium in Vibrio cholerae. However, when the cloned gene, present on plasmid pPM431, is introduced into E. coli K-12 this protein remains localized predominantly in the periplasmic space with no activity detected in the extracellular medium. Mutants of E. coli K-12 (tolA and tolB) which leak periplasmic proteins mimic excretion and release the haemolysin into the growth medium. Secretion of haemolysin into the periplasm is independent of perA (envZ) and in fact, mutants in perA (envZ) harbouring pPM431 show hyperproduction of periplasmic haemolysin. These results in conjunction with those for other V. cholerae extracellular proteins suggest that although E. coli K-12 can secrete these proteins into the periplasm, it lacks a specific excretion mechanism, present in V. cholerae, for the release of soluble proteins into the growth medium.
MeSH Terms
Animals
Cell Fractionation
Cloning, Molecular
Escherichia coli/genetics
Genes
Genes, Bacterial
Hemolysin Proteins/genetics,isolation & purification
Hemolysis
Plasmids
Sheep
Species Specificity
Subcellular Fractions/analysis
Transformation, Bacterial
Vibrio cholerae/genetics
Chemicals
Hemolysin Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Mercurio A
Manning P A
References (14)
14 references, click to expand
-
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):3962-72
PMID: 4555955
-
Molecular cloning of the tolC locus of Escherichia coli K-12 with the use of transposon Tn10.
Mol Gen Genet. 1981;184(3):430-3
PMID: 6278256
-
Secretion cloning vectors in Escherichia coli.
EMBO J. 1984 Oct;3(10):2437-42
PMID: 6094184
-
Molecular cloning of Vibrio cholerae enterotoxin genes in Escherichia coli K-12.
Proc Natl Acad Sci U S A. 1982 May;79(9):2976-80
PMID: 7045877
-
Purification and characterization of a hemolysin produced by Vibrio cholerae biotype El Tor: another toxic substance produced by cholera vibrios.
Infect Immun. 1979 Dec;26(3):1020-7
PMID: 118926
-
Pleiotropic properties and genetic organization of the tolA,B locus of Escherichia coli K-12.
J Bacteriol. 1972 Oct;112(1):74-83
PMID: 4627928
-
Mechanism of incorporation of cell envelope proteins in Escherichia coli.
Annu Rev Microbiol. 1982;36:435-65
PMID: 6756294
-
Cloning and functional characterization of the plasmid-encoded hemolysin determinant of Escherichia coli.
J Bacteriol. 1982 Sep;151(3):1290-8
PMID: 7050085
-
Escherichia coli pleiotropic mutant that reduces amounts of several periplasmic and outer membrane proteins.
J Bacteriol. 1979 Oct;140(1):229-39
PMID: 387722
-
Cloning of the structural gene (hly) for the haemolysin of Vibrio cholerae El Tor strain 017.
Gene. 1984 Nov;31(1-3):225-31
PMID: 6098527
-
The tolC locus of Escherichia coli affects the expression of three major outer membrane proteins.
J Bacteriol. 1982 Jun;150(3):1016-23
PMID: 6281230
-
Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands.
FEBS Lett. 1975 Oct 15;58(1):254-8
PMID: 773686
-
Defective transport and other phenotypes of a periplasmic "leaky" mutant of Escherichia coli K-12.
J Bacteriol. 1979 Nov;140(2):351-8
PMID: 387731
-
Non-O1 Vibrio cholerae hemolysin: purification, partial characterization, and immunological relatedness to El Tor hemolysin.
Infect Immun. 1984 Jul;45(1):192-6
PMID: 6429044