TcpC inhibits neutrophil extracellular trap formation by enhancing ubiquitination mediated degradation of peptidylarginine deiminase 4

  • Nat Commun. 2021 Jun 9;12(1):3481. doi: 10.1038/s41467-021-23881-8.
Qian Ou   #  1  2 ,  Jia-Qi Fang   #  1  2 ,  Zhe-Sheng Zhang   #  1  3 ,  Zhe Chi  1  2 ,  Jie Fang  1  3 ,  Di-Yan Xu  1  3 ,  Kai-Zhong Lu  1  3 ,  Meng-Qing Qian  1  3 ,  Da-Yong Zhang  1  3 ,  Jun-Ping Guo  1  3 ,  Wei Gao  1  3 ,  Na-Ru Zhang  1  3 ,  Jian-Ping Pan  4  5
Affiliations
  • 1. Institute of Translational Medicine, Zhejiang University City College, Hangzhou, P. R. China.
  • 2. Department of Basic Medical Sciences, Zhejiang University School of Medicine, Hangzhou, P. R. China.
  • 3. Department of Clinical Medicine, Zhejiang University City College School of Medicine, Hangzhou, P. R. China.
  • 4. Institute of Translational Medicine, Zhejiang University City College, Hangzhou, P. R. China. [email protected].
  • 5. Department of Clinical Medicine, Zhejiang University City College School of Medicine, Hangzhou, P. R. China. [email protected].
  • # Contributed equally.
PMID: 34108482 DOI: 10.1038/s41467-021-23881-8
Abstract

TcpC is a multifunctional virulence factor of uropathogenic E. coli (UPEC). Neutrophil extracellular trap formation (NETosis) is a crucial Anti-infection mechanism of neutrophils. Here we show the influence of TcpC on NETosis and related mechanisms. We show NETosis in the context of a pyelonephritis mouse model induced by TcpC-secreting wild-type E. coli CFT073 (CFT073wt) and LPS-induced in vitro NETosis with CFT073wt or recombinant TcpC (rTcpC)-treated neutrophils are inhibited. rTcpC enters neutrophils through caveolin-mediated endocytosis and inhibits LPS-induced production of ROS, proinflammatory cytokines and protein but not mRNA levels of peptidylarginine deiminase 4 (PAD4). rTcpC treatment enhances PAD4 ubiquitination and accumulation in proteasomes. Moreover, in vitro ubiquitination kit analyses show that TcpC is a PAD4-targetd E3 ubiquitin-ligase. These data suggest that TcpC inhibits NETosis primarily by serving as an E3 Ligase that promotes degradation of PAD4. Our findings provide a novel mechanism underlying TcpC-mediated innate immune evasion.

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