1887

Abstract

We have constructed four deletion derivatives of the cloned gene. Plasmid pDD1, in which the last 10 amino acids of the DnaK protein have been replaced by three different amino acids derived from the pBR322 vector, was as effective as plasmid pKP31, from which it was derived, in restoring the ability of a null mutant, BB1553, to plate phage and to grow at high temperatures. The other three mutations, involving much larger deletions of the gene, did not restore the ability to plate phage or the ability to grow at high temperatures. Plasmid pKUC2, which contains the whole gene and its promoters, was capable of restoring the ability of BB1553 to plate phage but, surprisingly, it did not restore the ability to grow at high temperatures, even though it was shown that the DnaK protein was efficiently expressed in these cultures. By transposon mutagenesis and sub-cloning, we have shown the presence of a second gene in plasmid pKP31 which is required for high-temperature growth of BB1553. This gene, which we call , is presumably also defective in the null mutant BB1553. We have also demonstrated that the inability of K756 to grow above 43.5 °C is complemented by sub-clones which contain the gene, but not by plasmid pKUC2.

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1991-06-01
2021-05-14
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References

  1. Bardwell J. C. A., Craig E. A. 1984; Major heat shock gene of Drosophila and Escherichia coli heat-inducible dnaK gene are homologous. Proceedings of the National Academy of Sciences of the United States of America 81848–852
    [Google Scholar]
  2. Bukau B., Walker G. C. 1989a; Cellular defects caused by deletion of the E. coli dnaK gene indicate roles for heat shock protein in normal metabolism. Journal of Bacteriology 171:2337–2346
    [Google Scholar]
  3. Bukau B., Walker G. C. 1989b; ΔdnaK52 mutants of E. coli have defects in chromosome segregation and plasmid maintenance at normal growth temperatures. Journal of Bacteriology 171:6030–6038
    [Google Scholar]
  4. Bukau B., Walker G. C. 1990; Mutations altering heat shock specific subunit of RNA polymerase suppress major cellular defects of E. coli mutants lacking the DnaK chaperone. EMBO Journal 9:4027–4036
    [Google Scholar]
  5. Chak K.-F., James R. 1984; Localization and characterization of a gene on the ColE3-CA38 plasmid that confers immunity to colicin E8. Journal of General Microbiology 130:701–710
    [Google Scholar]
  6. Chang A. C. Y., Cohen S. N. 1978; Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. Journal of Bacteriology 134:1141–1156
    [Google Scholar]
  7. Ezaki B., Ogura T., Hironori N., Hiraga S. 1989; Involvement of DnaK protein in mini-F plasmid replication: temperature-sensitive seg mutations are located in the dnaK gene. Molecular and General Genetics 218:183–189
    [Google Scholar]
  8. Georgopoulos C. 1977; A new bacterial gene (groPC) which affects λ DNA replication. Molecular and General Genetics 151:35–39
    [Google Scholar]
  9. Georgopoulos C., Herskowitz I. 1971; Escherichia coli mutants blocked in lambda DNA synthesis. In The Bacteriophage Lambda553–564 Herschey A. D. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  10. Grossman A. D., Straus D. B., Walter W. A., Gross C. A. 1987; σ32 synthesis can regulate the synthesis of heat shock proteins in E. coli. Genes and Development 1:179–184
    [Google Scholar]
  11. Herendeen S. L., Van Bogelan R. A., Neidhardt F. C. 1979; Levels of major proteins of E. coli during growth at different temperatures. Journal of Bacteriology 139:185–194
    [Google Scholar]
  12. Ingolia T. D., Slater M. J., Craig E. A. 1982; Saccharomyces cerevisae contains a complex multigene family related to the major heat shock-inducible gene of Drosophila. Molecular Cell Biology 2:1388–1392
    [Google Scholar]
  13. Johnson C., Chandresekhar G. N., Georgopoulos C. 1989; E. coli DnaK and GrpE heat shock proteins interact both in vivo and in vitro. Journal of Bacteriology 171:1590–1596
    [Google Scholar]
  14. Kawasaki Y., Wada C., Yura T. 1990; Roles of E. coli heat shock proteins DnaK, DnaJ and GrpE in mini-F plasmid replication. Molecular and General Genetics 220:277–282
    [Google Scholar]
  15. Liberek K., Georgopoulos C., Zylicz M. 1988; Role of the DnaK and DnaJ heat shock proteins in the initiation of bacteriophage λ DNA replication. Proceedings of the National Academy of Sciences of the United States of America 856632–6636
    [Google Scholar]
  16. Lindquist S., Craig E. A. 1988; The heat shock proteins. Annual Review of Genetics 22:631–677
    [Google Scholar]
  17. Maniatis T., Fritsch E. F., Sambrook J. 1982; Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. Merrick M. J., Gibbins J. R., Postgate J. R. 1987; A rapid and efficient method of plasmid transformation of Klebsiella pneumoniae and Escherichia coli. Journal of General Microbiology 133:2053–2057
    [Google Scholar]
  19. Miller J. H. 1972; Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Neidhardt F. C., Van Bogelen R. A. 1981; Positive regulatory gene for temperature-controlled proteins in E. coli. Biochemical and Biophysical Research Communications 100:894–900
    [Google Scholar]
  21. Paek K.-H., Walker G. C. 1987; E. coli dnaK null mutants are inviable at high temperature. Journal of Bacteriology 169:283–290
    [Google Scholar]
  22. Saito H., Uchida H. 1977; Initiation of the DNA replication of bacteriophage lambda in E. coli K12. Journal of Molecular Biology 113:1–25
    [Google Scholar]
  23. Tilly K., Yarmolinsky M. 1989; Participation of E. coli heat shock proteins DnaJ, DnaK and GrpE in P1 plasmid replication. Journal of Bacteriology 171:6025–6029
    [Google Scholar]
  24. Vieira J., Messing J. 1982; The pUC plasmids, an M13mp7 derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
    [Google Scholar]
  25. Wadsworth S. C. 1982; A family of related proteins is encoded by the major Drosophila heat shock gene family. Molecular Cell Biology 2:286–292
    [Google Scholar]
  26. Wickner S. H. 1990; Three E. coli heat shock proteins are required for P1 plasmid DNA replication: formation of an active complex between E. coli DnaJ protein and the P1 initiator protein. Proceedings of the National Academy of Sciences of the United States of America 872690–2694
    [Google Scholar]
  27. Yamamori T., Yura T. 1982; Genetic control of heat-shock protein synthesis and its bearing on growth and thermal resistance in E. coli K12. Proceedings of the National Academy of Sciences of the United States of America 79860–864
    [Google Scholar]
  28. Zylicz M., Ang D., Georgopoulos C. 1987; The grpE protein of E. coli . Journal of Biological Chemistry 62:17437–17442
    [Google Scholar]
  29. Zylicz M., Ang D., Liberek K., Georgopoulos C. 1989; Initiation of A DNA replication with purified host and bacteriophage-encoded proteins: the role of the DnaK, DnaJ and GrpE heat shock proteins. EMBO Journal 8:1601–1608
    [Google Scholar]
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