Functional characterization of NLRX1 in rabbit during enterohemorrhagic Escherichia coli infection.

Nucleotide oligomerization domain (NOD) like receptor X1 (NLRX1) is a member of pattern recognition receptor, which has been linked to viral response, cancer, and inflammatory diseases. In this study, rabbit NLRX1 (rNLRX1) was firstly cloned from RK-13 cells, which protein contained a NACHT domain and seven LRRs. rNLRX1 was widely expressed in tissues of rabbits, and highly increased in liver, spleen, kidney, and colon after infected with enterohemorrhagic Escherichia coli (EHEC). Overexpression of rNLRX1 negatively regulated NF-κB signaling, and impaired the expression of pro-inflammatory cytokines and defensins. Moreover, deficient of rNLRX1 in RK-13 cells was performed to investigate the possible roles of rNLRX1. Upon EHEC stimulation, knockdown of rNLRX1 markedly enhanced NF-κB activation and downstream responsive cytokines (IL1β and TNFα) and β-defensins (DEFB114 , DEFB124 , and DEFB125). Furthermore, overexpression of rNLRX1 promoted the proliferation of EHEC, whereas knockdown of rNLRX1 inhibited its growth. Our study identified that rNLRX1 acts as a negative regulatory in anti-microbial responses after EHEC infection. • rNLRX1 contained a NACHT domain and seven LRRs. • rNLRX1 highly increased after infected with enterohemorrhagic Escherichia coli. • rNLRX1 negatively regulated NF-κB signaling. • rNLRX1 impaired the expression of pro-inflammatory cytokine and defensins. • Knockdown of rNLRX1 markedly enhanced NF-κB activation. [ABSTRACT FROM AUTHOR]