Role of Region C in Regulation of the Heat Shock Gene-Specific Sigma Factor of Escherichia coli , ς 32 - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Journal of Bacteriology Année : 1999

Role of Region C in Regulation of the Heat Shock Gene-Specific Sigma Factor of Escherichia coli , ς 32

Résumé

ABSTRACT Expression of heat shock genes is controlled in Escherichia coli by the antagonistic action of the ς 32 subunit of RNA polymerase and the DnaK chaperone system, which inactivates ς 32 by stress-dependent association and mediates ς 32 degradation by the FtsH protease. A stretch of 23 residues (R122 to Q144) conserved among ς 32 homologs, termed region C, was proposed to play a role in ς 32 degradation, and peptide analysis identified two potential DnaK binding sites central and peripheral to region C. Region C is thus a prime candidate for mediating stress control of ς 32 , a hypothesis that we tested in the present study. A peptide comprising the central DnaK binding site was an excellent substrate for FtsH, while a peptide comprising the peripheral DnaK binding site was a poor substrate. Replacement of a single hydrophobic residue in each DnaK binding site by negatively charged residues (I123D and F137E) strongly decreased the binding of the peptides to DnaK and the degradation by FtsH. However, introduction of these and additional region C alterations into the ς 32 protein did not affect ς 32 degradation in vivo and in vitro or DnaK binding in vitro. These findings do not support a role for region C in ς 32 control by DnaK and FtsH. Instead, the ς 32 mutants had reduced affinities for RNA polymerase and decreased transcriptional activities in vitro and in vivo. Furthermore, cysteines inserted into region C allowed cysteine-specific cross-linking of ς 32 to RNA polymerase. Region C thus confers on ς 32 a competitive advantage over other ς factors to bind RNA polymerase and thereby contributes to the rapidity of the heat shock response.

Dates et versions

hal-04267936 , version 1 (02-11-2023)

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Florence Arsène, Toshifumi Tomoyasu, Axel Mogk, Christiane Schirra, Agnes Schulze-Specking, et al.. Role of Region C in Regulation of the Heat Shock Gene-Specific Sigma Factor of Escherichia coli , ς 32. Journal of Bacteriology, 1999, 181 (11), pp.3552-3561. ⟨10.1128/JB.181.11.3552-3561.1999⟩. ⟨hal-04267936⟩
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