| Abstract |
Escherichia coli synthesizes curli amyloid fibers extracellularly during biofilm formation and host colonization. The proteostasis network regulates the major curli subunit, CsgA, to prevent intracellular amyloid aggregation, yet the degradation mechanism remains elusive. Here, through a comprehensive investigation employing genetically engineered E. coli, multi-copy-suppressor screening, and biochemical analyses, we identify periplasmic serine protease Prc as a key player in CsgA degradation. Prc directly degrades CsgA through internal cleavage, differing from canonical tail-specific proteases. Although the bacterial HtrA homologs DegP and DegQ exhibit limited CsgA degradation activity in vitro in the presence of the suicide activator YjfN, deletion of these proteases did not affect native CsgA degradation in vivo. Instead, Prc, in coordination with the periplasmic chaperone CsgC, prevents the periplasmic accumulation of CsgA amyloid-like aggregates. Furthermore, impairment of efficient secretion and proteolytic systems leads to reduced csg operon expression mediated by the Rcs and Cpx two-component systems. Our findings reveal a dual-layered strategy employed by E. coli to prevent intracellular accumulation of extracellular amyloids at both protein degradation and transcriptional regulation levels.
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