YaeL (EcfE) activates the ςE pathway of stress response through a site-2 cleavage of anti-ςE, RseA

Abstract

Escherichia coli YaeL (EcfE) is a homolog of human site-2 protease (S2P), a membrane-bound zinc metalloprotease involved in regulated intramembrane proteolysis. We have shown previously that YaeL, having essential metalloprotease active site motifs in the cytoplasmic domain, is indispensable for viability. Here, we obtainedrpoE, encoding an extracytoplasmic stress response ς factor (ςE), as a multicopy suppressor against the yaeLdisruption. Whereas ςE is thought to be activated by regulated cleavage of RseA on the periplasmic side by the DegS protease, we found that a degradation intermediate of RseA consisting of the transmembrane and the cytoplasmic domains accumulated in the YaeL-depleted cells. This intermediate was degraded on expression of YaeL but not of its metalloprotease motif mutants. Cells depleted of YaeL were incapable of activating a ςE-dependent promoter in response to an envelope stress. It is suggested that ςEactivation involves two successive proteolytic cleavages: first, at a periplasmic site by DegS; second, at a cytoplasmic or intramembrane site by YaeL. Thus, YaeL is positively required for the ςEextracytoplasmic stress response.