title: Response of Escherichia coli cell membranes to induction of lambda cl857 prophage by heat shock.
abstract: Heat shock induces protein aggregation in Escherichia coli and E. coli (lambda cl857). The aggregates (S fraction) appear 15 min post-induction and are separable from membranes by sucrose density-gradient centrifugation. The S fraction quickly disappears in wild type strains but persists in rpoH mutant with concomitant quick inner membrane destruction. We propose that: (1) the disappearance of the S fraction reflects a rpoH-dependent processing, (2) the membrane destruction explains the lethality of the rpoH mutation at elevated temperatures; and (3) the protection of the inner membrane integrity is an important physiological function of the heat-shock response. We assume that the S fraction of aggregated proteins represents the signal inducing the heat-shock response. The prophage thermo-induction results in an increase (35 min post-induction) in the A fraction resembling that of the adhesion zones of the membranes. This fraction is greater than the corresponding fraction from uninduced cells. The increase is mediated by the lambda late genes, since it is absent in the induced E. coli (lambda cl857 Qam21). Since heat shock is widely used for induction of the lambda promoters in expression vectors it is possible that the formation of the protein aggregates (though transient in WT strains) and/or the fragility of membranes in rpoH mutants may be the cause of poor expression of cloned genes or may lead to mistaken localization of their expression products.
authors.affiliation: Department of Biochemistry, University of Gdańsk, Poland.
authors.forename: K
authors.initials: K
authors.lastname: Kucharczyk
authors.forename: E
authors.initials: E
authors.lastname: Laskowska
authors.forename: A
authors.initials: A
authors.lastname: Taylor
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D010582:Bacteriophage lambda / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology*; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000378:metabolism / Q000502:physiology*; D008565:Membrane Proteins / Q000502:physiology; D033903:Periplasmic Proteins; D012697:Serine Endopeptidases / Q000378:metabolism; D013347:Subcellular Fractions / Q000502:physiology; D014764:Viral Proteins / Q000378:metabolism; D014775:Virus Activation / Q000502:physiology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C068113:GrpE protein, Bacteria; C489400:GrpE protein, E coli; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D008565:Membrane Proteins; D033903:Periplasmic Proteins; C066119:Q protein, Bacteriophage lambda; D014764:Viral Proteins; C061682:DegP protease; D012697:Serine Endopeptidases; C063842:dnaK protein, E coli