E. coli Mutants Lacking the dnaK Heat Shock Gene: Identification of Cellular Defects and Analysis of Suppressor Mutations

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Heat Shock
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Abstract

We investigated the role of the DnaK (HSP70) ehaperone In metabolism of Escherichia coli by analyzing cellular defects caused by deletion of the dnaK gene (ΔdnaK52). ΔdnaK52 mutants are cold sensitive as well as temperature sensitive and thus possess a very narrow temperature range for growth. At intermediate (30°C) temperature, ΔdnaK52mutants display severe defects in major cellular processes such as cell division, chromosome segregation, replication of low copy number plasmids and regulation of heat shock gene expression that lead to poor growth and genetic instability of the cells. These results indicate important functions of DnaK in cellular metabolism of E. coli at a wide range of growth temperatures.

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References

  • Bardwell, JCA and Craig, EA, (1984) Major heat shock gene of Drosophila and the Escherichia coli heat-indueible dnaK gene are homologous. Proc. Natl. Acad. Sci. USA 81: 848–852.

    Article  Google Scholar 

  • Bukau, B and Walker, GC, (1989a) Cellular defects caused by deletion of the Escherichia coli dnaK gene indicate roles for heat shock protein in normal metabolism. J. Bacteriol., 171: 2337–2346.

    PubMed  CAS  Google Scholar 

  • Bukau, B and Walker, GC, (1989b) ΔdnaK52 mutants of Escherichia coli have defects in chromosome segregation and plasmid maintenance at normal growth temperatures. J. Bacteriol., 171: 6030–6038.

    CAS  Google Scholar 

  • Bukau, B and Walker, GC, Mutations altering heat shock specific subunit of RNA polymerase suppress major celltuar defects of E. coli mutants lacking the DnaK chaperone. EMBO J., in press.

    Google Scholar 

  • Bukau, B, Donnelly, CE and Walker, GC, (1989) DnaK and GroE proteins play roles in E. coli metabolism at low and intermediate temperatures as well as at high temperatures, p. 27–36. In Pardue, ML, Feramiseo, JR and Lindquist, S, eds., Stress Induced Proteins. Alan. R Liss, Inc., New York.

    Google Scholar 

  • Chiang, H-L, Terlecky, SR, Plant, CP and Dice, JF, (1989) A role for a 70- kilodalton heat shock protein in lysosomal degradation of intracellular proteins. Science, 246: 382–385.

    Article  PubMed  CAS  Google Scholar 

  • Echols, H, (1986) Multiple DNA-protein interactions governing high-precision DNA transactions. Science, 233: 1050–1056.

    Article  PubMed  CAS  Google Scholar 

  • Funnell, BE, (1988) Mini-Pi plasmid partitioning: excess ParB protein destabilizes plasmids containing the centromereparS. J. Baeteriol., 170: 954–960.

    CAS  Google Scholar 

  • Grossman, AD, Straus, DB, Walter, WA and Gross, CA (1987) σ32 synthesis can regulate the synthesis of heat shock proteins in Escherichia coli. Genes Dev., 1: 179–184.

    Article  PubMed  CAS  Google Scholar 

  • Hirota, Y, Ryter, A and Jacob, F, (1968) Thermosensitive mutants of E. coli affected in the processes of DNA synthesis and cellular division. Cold Spring Harbor Symp. Quant. Biol., 33: 677–694.

    PubMed  CAS  Google Scholar 

  • Itikawa, H and Ryu, J-I, (1979) Isolation and characterization of a temperature- sensitive dnaK mutant of Escherichia coli B. J. Baeteriol., 138: 339–344.

    CAS  Google Scholar 

  • Liberek, K, Georgopoulos, C and Zylicz, M, (1988) Role of the Escherichia coli DnaK and DnaJ heat shock proteins in the initiation of bacteriophage X DNA replication. Proc. Natl. Acad. Sci. USA, 85: 6632–6636.

    Article  CAS  Google Scholar 

  • Lindquist, Sand Craig, EA, (1988) The heat-shock proteins. Annu. Rev. Genet. 22: 631–677.

    Article  PubMed  CAS  Google Scholar 

  • Morimoto, R I, Tissieres, A and Georgopoulos, C, eds. (1990) Stress Proteins in Biology and Medicine. Cold Spring Harbor monograph Series Vol. 19, Cold Spring Harbor Press, Cold Spring Harbor, N.Y

    Google Scholar 

  • Sakakibara, Y, (1988) The dnaK gene of Escherichia coli functions in initiation of chromosome replication. J. Baeteriol., 170: 972–979.

    CAS  Google Scholar 

  • Sell, SM, Eisen, C, Ang, D, Zylicz, M and Georgopoulos, C, (1990) Isolation and characterization of dnaJ null mutants of Escherichia coli J. Bacteriol., 172: 4827–4835.

    PubMed  CAS  Google Scholar 

  • Sternberg, N and Austin, S, (1983) Isolation and characterization of PI minireplieons, λ-P1:5R and λ,-P1:5L. J. Bacteriol., 153: 800–812.

    PubMed  CAS  Google Scholar 

  • Straus, DB, Walter, WA and Gross, CA (1987) The heat shock response of E. coli is regulated by changes in the concentration of σ32. Nature, 329: 348–351.

    Article  PubMed  CAS  Google Scholar 

  • Straus, DB, Walter, WA and Gross, CA, (1988) Escherichia coli heat shock gene mutants are defective in proteolysis. Genes Dev., 2: 1851–1858.

    Article  PubMed  CAS  Google Scholar 

  • Tilly, K, McKittrick, N, Zylicz, M and Georgopoulos, C, (1983) The dnaK protein modulates the heat shock response of Escherichia coli. Cell, 34: 641–646.

    Article  PubMed  CAS  Google Scholar 

  • Tilly, K, Spence, J and Georgopoulos, C, (1989) Modulation of stability of the Escherichia coli heat shock regulatory factor σ32. J. Bacteriol., 171: 1585–1589.

    PubMed  CAS  Google Scholar 

  • Tilly, K and Yarmolinsky, M, (1989) Participation of Escherichia coli heat shock proteins DnaK, DnaJ, and GrpE in PI plasmid replication. J. Bacteriol., 171: 6025–6029.

    PubMed  CAS  Google Scholar 

  • Zylicz, M, Ang, D, Liberek, Kand Georgopoulos, C, (1989) Initiation of lambda DNAreplication with purified host- and baeteriophage-encoded proteins: the role of the dnaK, dnaJ and grpE heat shock proteins. EMBO J., 8: 1601–1608.

    PubMed  CAS  Google Scholar 

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© 1991 Springer-Verlag Berlin Heidelberg

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Bukau, B., Walker, G.C. (1991). E. coli Mutants Lacking the dnaK Heat Shock Gene: Identification of Cellular Defects and Analysis of Suppressor Mutations. In: Maresca, B., Lindquist, S. (eds) Heat Shock. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-76679-4_6

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  • DOI: https://doi.org/10.1007/978-3-642-76679-4_6

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-76681-7

  • Online ISBN: 978-3-642-76679-4

  • eBook Packages: Springer Book Archive

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