ATG9 inhibits Rickettsia binding to the host cell surface by blocking the rOmpB-XRCC6/KU70 interaction

ickettsiae are tick-borne pathogens that infect human hosts through poorly characterized mechanisms. Herein, we report that ATG9 (Autophagy related 9) plays a previously unrecognized role in inhibiting Rickettsia binding to the host cell surface. Unexpectedly, this new function of ATG9 is likely independent of macroautophagy/Autophagy. Instead, ATG9 acts as a host defending factor by binding to XRCC6/KU70, a receptor of the Rickettsia outer-membrane protein rOmpB. Both ATG9 and rOmpB bind to the DNA-binding domain of XRCC6, suggesting a competitive role for ATG9 occupying the binding site of rOmpB to abrogate Rickettsia binding. Furthermore, we show that rapamycin transcriptionally activates ATG9 and inhibits rOmpB-mediated Infection in a mouse model. Collectively, our study reveals a novel innate mechanism regulating Rickettsia Infection and suggests that agonists of ATG9 May be useful for developing therapeutic strategies for the intervention of rickettsial diseases.Abbreviation: APEX2: apurinic/apyrimidinic endodeoxyribonuclease 2; ATG: Autophagy related; BafA1: bafilomycin A1; CQ: chloroquine; E. coli: Escherichia coli; GST: glutathione S-transferase; ICM: immunofluorescence confocal microscopy; IP-Mass: immunoprecipitation-mass spectrometry; KD: knockdown; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MTOR: mechanistic target of rapamycin kinase; rOmpB: rickettsial outer membrane protein B; SAP: SAF-A/B, Acinus, and PIAS; SQSTM1/p62: sequestosome 1; TEM: transmission electron microscopy; TFEB: transcription factor EB; VWA: von Willebrand factor A; XRCC6/KU70: X-ray repair cross complementing 6.

Autophagy-related gene; host defense; infectious disease; rOmpB; rickettsia; tick-borne pathogen.