1887

Abstract

The cellular level of the heat-shock sigma factor RpoH ( ) is negatively controlled by chaperone-mediated proteolysis through the essential metalloprotease FtsH. Point mutations in the highly conserved region 2.1 stabilize RpoH . To assess the importance of this turnover element, hybrid proteins were constructed between RpoH and RpoH, a stable RpoH protein that differs from region 2.1 of RpoH at several positions. Nine amino acids forming a putative -helix were exchanged between the two proteins. Both hybrids were active sigma factors and showed intermediate protein stability. Introduction of RpoH region 2.1 into the general stress sigma factor RpoS, which is a substrate of the ClpXP protease, did not render RpoS susceptible to FtsH. Hence, region 2.1 alone is not sufficient to confer FtsH sensitivity to other proteins. Region 2.1 is not a major chaperone-binding site since DnaK and DnaJ bound efficiently to all RpoH variants. The stability of the mutated RpoH proteins correlated with their stability in a purified degradation system, suggesting that region 2.1 might be directly involved in the interaction with the FtsH protease.

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2007-08-01
2019-12-09
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