Complementation of the hha mutation in Escherichia coli by the ymoA gene from Yersinia enterocolitica: dependence on the gene dosage Free

Abstract

The Hha protein from is highly similar (82%) to the YmoA protein from . Both are members of a new class of proteins that modulates gene expression, probably by influencing DNA topology. In this paper, complementation of the mutation in by the gene from has been studied. We show that the gene complements one of the phenotypic properties of mutants (high level of haemolysin production when they carry the recombinant plasmid pANN202-312) when cloned in a medium-copy-number plasmid but not when carried in a low-copy-number plasmid. Western blot analysis of the expression of YmoA in rules out inefficient expression of the protein. Surprisingly, the gene itself fails to complement the mutation when cloned in a medium-copy-number vector and causes genetic rearrangements of the chromosome as a consequence of insertion sequences mobilization.

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1996-07-01
2024-03-29
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References

  1. Blanco A., Juárez A., Pastor F. I. J. 1991; Overproduction of a Clostridium cellulolyticum endoglucanase by mutant strains of Escherichia coli. FEMS Microbiol Eett 81:221–226
    [Google Scholar]
  2. Bolivar F., Rodríguez R.L., Green P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. 1977; Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113
    [Google Scholar]
  3. Carmona M., Balsalobre C., Muñoa, F., Mouriño M., Jubete Y., De la Cruz F., Juárez A. 1993; Escherichia coli hha mutants, DNA supercoiling and expression of the haemolysin genes from the recombinant plasmid pANN202-312. Mol Microbiol 9:1011–1018
    [Google Scholar]
  4. Claverie-Martfn F., Diaz-Torres M.R., Yancey S. D., Kushner S. K. 1989; Cloning of the altered mRNA stability (ams) gene of Escherichia coli K-12. J Bacteriol 171:5479–5486
    [Google Scholar]
  5. Cornelis G. R., Sluiters C., Delor I., Gelb D., Kaninga K., Lambert de Rouvrait C., Sory M. P., Vanooteghem J. C., Michaelis T. 1991; ymo A, a Yersinia enterocolitica chromosomal gene modulating the expression of virulence functions. Mol Microbiolb1023–1034
    [Google Scholar]
  6. Cunningham T. P., Montelaro R. C., Rushlow K. E. 1993; Lentivirus envelope sequences and proviral genomes are stabilized in Escherichia coli when cloned in low-copy-number plasmid vectors. Gene 9:93–98
    [Google Scholar]
  7. De la Cruz F., Carmona M., Juárez A. 1992; The Hha protein from Escherichia coli is highly homologous to the Ymo A protein from Yersinia enterocolitica. Mol Microbiol 6:3451–3452
    [Google Scholar]
  8. Godessart N., Muñoa F. J., Regué M., Juárez A. 1988; Chromosomal mutations that increase the production of a plasmid- encoded haemolysin in Escherichia coli. J Gen Microbiol 134:2779–2787
    [Google Scholar]
  9. Goebel W., Hedgepth J. 1982; Cloning and functional characterization of the plasmid-encoded haemolysin determinant of Escherichia coli. J Bacteriol 151:1290–1298
    [Google Scholar]
  10. Hulton C.S.J., Seirafi A., Hinton J. C. D., Sidebotham J. M., Waddell L., Pawit G. D., Owen-Hughes T., Spassky A., Bue H., Higgins C. F. 1990; Histone-like protein HI (H-NS), DNA supercoiling and gene expression in bacteria. Cell 63:631–642
    [Google Scholar]
  11. Mikulskis A.V., Cornelis G. R. 1994; A new class of proteins regulating gene expression in enterobacteria. Mol Microbiol 11:77–86
    [Google Scholar]
  12. Nieto J. M., Tomás J, Juárez A. 1987; Secretion of an Aeromonas hydrophila aerolysin by a mutant strain of Escherichia coli. FEMS Microbiol Lett 48:413–417
    [Google Scholar]
  13. Nieto J. M., Carmona M., Bolland S., Jubete Y., De la Cruz F., Juárez A. 1991; The hha gene modulates haemolysin expression in Escherichia coli. Mol Microbiol 5:1285–1293
    [Google Scholar]
  14. Simon R., Hötte B., Klauke B., Kosier B. 1991; Isolation and characterization of insertion sequence elements from gram-negative bacteria by using new broad-host-range, positive selection vectors. J Bacteriol 173:1502–1508
    [Google Scholar]
  15. Spratt B. G., Boyd A., Stoker N. 1980; Defective and plaque- forming lambda transducing bacteriophage carrying penicillin- binding protein-cell shape genes : genetic and physical mapping and identification of gene products from the lip-dac A-rod A-pbp A-leu S region of the Escherichia coli chromosome. J Bacteriol 143:569–581
    [Google Scholar]
  16. Spurio R., Dürrenberger M., Falconi M., La Teana A., Pon C. L., Guaterei C. O. 1992; Lethal overproduction of the Escherichia coli nucleoid protein H-NS: ultramicroscopic and molecular autopsy. Mol Gen Genet 231:201–211
    [Google Scholar]
  17. Ueguchi C., Ito K. 1992; Multicopy suppression: an approach to understanding intracellular functioning of the protein export system. J Bacteriol 174:1454–1461
    [Google Scholar]
  18. Wang R.F., Kushner S. R. 1991; Construction of versatile low- copy-number vectors for cloning, sequencing and gene expression in Escherichia coli. Gene 100:195–199
    [Google Scholar]
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