1887

Abstract

Previously reported cell fractionation experiments have yielded conflicting information on the cellular localization of the DnaK heat shock protein of . Here we used immunogold labelling of ultra-thin sections to determine the localization of DnaK in unstressed cells at 30 °C as well as in heat-shocked cells. In cells grown at 30 °C, gold particles were found predominantly in the cytoplasm, indicating that the majority of the DnaK molecules are cytoplasmic; however, a fraction of the gold particles was located in proximity to the membranes, raising the possibility that a subpopulation of DnaK proteins is membrane-associated. Heat shock of the cells did not induce detectable relocalization of DnaK.

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1993-01-01
2024-04-20
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References

  1. Bardwell J. C. A., Craig E. A. 1984; Major heat shock gene of Drosophila and the Escherichia coli heat-inducible dnaK gene are homologous. Proceedings of the National Academy of Sciences of the United States of America 81:848–852
    [Google Scholar]
  2. Bardwell J. C. A., Tilly K., Craig E., King J., Zyliyz M., Georgopoulos C. 1986; The nucleotide sequence of the Escherichia coli K12 dnaJ+gene. Journal of Biological Chemistry 261:1782–1785
    [Google Scholar]
  3. Bukau B., Walker G. C. 1989a; Cellular defects caused by deletion of the Escherichia colidnaK gene indicate roles for heat shock protein in normal metabolism. Journal of Bacteriology 171:2337–2346
    [Google Scholar]
  4. Bukau B., Walker G. C. 1989b; ΔdnaK52 mutants of Escherichia coli have defects in chromosome segregation and plasmid maintenance at normal growth temperatures. Journal of Bacteriology 171:6030–6038
    [Google Scholar]
  5. 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 DnaJ chaperone. EMBO Journal 9:4027–4036
    [Google Scholar]
  6. Francesconi S. C., Macalister T. J., Setlow B., Setlow P. 1988; Immunoelectron microscopic localization of small, acid- soluble spore proteins in sporulating cells of Bacillus subtilis. Journal of Bacteriology 170:5963–5967
    [Google Scholar]
  7. Gething M.-J., Sambrook J. 1992; Protein folding in the cell.. Nature; London: 355:33–45
    [Google Scholar]
  8. Guidon P. T.Jr Hightower L. E. 1986; Purification and initial characterization of the 71-kilodalton rat heat-shock protein and its cognate as fatty acid binding proteins. Biochemistry 25:3231–3239
    [Google Scholar]
  9. Jacq A., Kern R., Tsugita A., Kohiyama M. 1989; Purification and characterization of a low-molecular-weight membrane protein with affinity for the Escherichia coli origin of replication. Journal of Bacteriology 171:1409–1416
    [Google Scholar]
  10. Kostyal D. A., Farrell M., Mccabe A., Mei Z., Firshein W. 1989; Replication of an RK2 miniplasmid derivative in vitro by a DNA/membrane complex extracted from Escherichia coli: involvement of the dnaA but not dnaK host proteins and association of these and plasmid-encoded proteins with the inner membrane. Plasmid 21:226–237
    [Google Scholar]
  11. Liberek K., Georgopoulos C., Zylicz M. 1988; Role of the Escherichia coli DnaK and DnaJ heat shock proteins in the initiation of bacteriophage λDNA replication. Proceedings of the National Academy of Sciences of the United States of America 85:6632–6636
    [Google Scholar]
  12. Miller J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor NY; Cold Spring Harbor Laboratory:
    [Google Scholar]
  13. Morimoto R. I., Tissieres A., Georgopoulos C.editors 1990 Stress Proteins in Biology and Medicine Cold Spring Harbor monograph Series Vol. 19) Cold Spring Harbor NY; Cold Spring Harbor Laboratory:
    [Google Scholar]
  14. Paek K. -H., Walker G. C. 1987; Escherichia coli dnaK null mutants are inviable at high temperature. Journal of Bacteriology 169:283–290
    [Google Scholar]
  15. Schlossman D. M., Schmid S. L., Braell W. A., Rothman J. E. 1984; An enzyme that removes clathrin coats: purification of an uncoating ATPase. Journal of Cellular Biology 99:723–733
    [Google Scholar]
  16. Simon G. T., Thomas J. A., Chorneyko K. A., Carlemalm E. 1987; Rapid embedding in Lowicryl K4M for immunoelectron microscopic studies. . Journal of Electron Microscopical Techniques 6:317–324
    [Google Scholar]
  17. Tommassen J., Leunissen J., Van Damme-Jongsten M., Overduin P. 1985; Failure of E. coli K-12 to transport PhoE-LacZ hybrid proteins out of the cytoplasm. EMBO Journal 4:1041–1047
    [Google Scholar]
  18. Zylicz M., Nieradko J., Taylor K. 1983; Escherichia colidnaJ- and dnaK-gene products: synthesis in minicells and membrane-affinity. Biochemical and Biophysical Research Communications 110:176–180
    [Google Scholar]
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