title: Ehrlichia type IV secretion effector ECH0825 is translocated to mitochondria and curbs ROS and apoptosis by upregulating host MnSOD.
abstract: Ehrlichia chaffeensis infects monocytes/macrophages and causes human monocytic ehrlichiosis. To determine the role of type IV secretion (T4S) system in infection, candidates for T4S effectors were identified by bacterial two-hybrid screening of E. chaffeensis hypothetical proteins with positively charged C-terminus using E. chaffeensis VirD4 as bait. Of three potential T4S effectors, ECH0825 was highly upregulated early during exponential growth in a human monocytic cell line. ECH0825 was translocated from the bacterium into the host-cell cytoplasm and localized to mitochondria. Delivery of anti-ECH0825 into infected host cells significantly reduced bacterial infection. Ectopically expressed ECH0825 also localized to mitochondria and inhibited apoptosis of transfected cells in response to etoposide treatment. In double transformed yeast, ECH0825 localized to mitochondria and inhibited human Bax-induced apoptosis. Mitochondrial manganese superoxide dismutase (MnSOD) was increased over ninefold in E. chaffeensis-infected cells, and the amount of reactive oxygen species (ROS) in infected cells was significantly lower than that in uninfected cells. Similarly, MnSOD was upregulated and the ROS level was reduced in ECH0825-transfected cells. These data suggest that, by upregulating MnSOD, ECH0825 prevents ROS-induced cellular damage and apoptosis to allow intracellular infection. This is the first example of host ROS levels linked to a bacterial T4S effector.
authors.affiliation: Department of Veterinary Biosciences, The Ohio State University, 1925 Coffey Road, Columbus, OH 43210, USA.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Liu
authors.forename: Weichao
authors.initials: W
authors.lastname: Bao
authors.forename: Mingqun
authors.initials: M
authors.lastname: Lin
authors.forename: Hua
authors.initials: H
authors.lastname: Niu
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Rikihisa
mesh_terms: D017209:Apoptosis; D001426:Bacterial Proteins / Q000378:metabolism; D058947:Bacterial Secretion Systems; D002460:Cell Line; D018469:Ehrlichia chaffeensis / Q000472:pathogenicity*; D006801:Humans; D008928:Mitochondria / Q000201:enzymology / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism / Q000382:microbiology; D021381:Protein Transport; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D017382:Reactive Oxygen Species; D037521:Virulence Factors; D013482:Superoxide Dismutase