title: Recombinant YopJ induces apoptosis in murine peritoneal macrophages in vitro: involvement of mitochondrial death pathway.
abstract: Yersinia species during infection adhere to host immune cells primarily to macrophages and employ its secretary proteins known as Yersinia outer proteins to trigger death in infected cells. In the present study, it is shown that recombinant Yersinia outer protein J (rYopJ) could induce apoptosis in murine peritoneal macrophages in vitro as assessed by morphological features, internucleosomal DNA fragmentation, change in mitochondrial membrane potential (MMP) (Deltapsim), activation of caspases and Annexin V binding. rYopJ-induced cell death was dose and time dependent. Pre-treatment with broad-spectrum caspase inhibitor Z-VAD-FMK, caspase-3 inhibitor Ac-DEVD-CHO and caspase-8 inhibitor Z-IETD-FMK prevented the change in MMP and DNA fragmentation, suggesting caspase-dependent apoptosis of rYopJ-treated macrophages. Blocking the endocytosis by pre-treatment of cells with cytochalasin B did not prevent the rYopJ-induced macrophages apoptosis. The data further suggest that rYopJ-induced apoptosis is mediated by molecules upstream of caspase-8 and relay through mitochondrial pathway involving Bax, Bcl-2, activation of caspase-8 and caspase-3, Bid and polyadenosine diphosphate-ribose polymerase cleavage, cytochrome c release and DNA fragmentation.
authors.affiliation: School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi, India.
authors.forename: Ashok Kumar
authors.initials: AK
authors.lastname: Pandey
authors.forename: Ajit
authors.initials: A
authors.lastname: Sodhi
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017304:Annexin A5 / Q000276:immunology / Q000378:metabolism; D017209:Apoptosis / Q000276:immunology*; D053477:Apoptotic Protease-Activating Factor 1 / Q000819:agonists / Q000276:immunology / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D053148:Caspase 3 / Q000276:immunology / Q000378:metabolism; D053181:Caspase 8 / Q000276:immunology / Q000378:metabolism; D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D045304:Cytochromes c / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000276:immunology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008928:Mitochondria / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology; D010930:Plague / Q000276:immunology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D015007:Yersinia / Q000276:immunology*; D051028:bcl-2-Associated X Protein / Q000819:agonists / Q000276:immunology / Q000378:metabolism
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D017304:Annexin A5; C506593:Apaf1 protein, mouse; D053477:Apoptotic Protease-Activating Factor 1; D001426:Bacterial Proteins; C494827:Bax protein, mouse; D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D009842:Oligopeptides; D019253:Proto-Oncogene Proteins c-bcl-2; D011994:Recombinant Proteins; C109227:YopP protein, Yersinia; C099158:acetyl-aspartyl-glutamyl-valyl-aspartal; D051028:bcl-2-Associated X Protein; C403753:benzyloxycarbonyl-isoleucyl-glutamyl-threonyl-aspartic acid fluoromethyl ketone; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D003571:Cytochalasin B; D045304:Cytochromes c; D053148:Caspase 3; D053181:Caspase 8