Periplasmic serine protease Prc is responsible for amyloid subunit CsgA degradation and proteostasis in Escherichia coli

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. Additionally, disruptions in secretion efficiency and proteolytic systems reduce csg operon expression through activation of the Rcs and Cpx two-component systems. These 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. This study provides insights into the mechanisms ensuring cellular homeostasis during curli biogenesis.