title: Mitochondria-targeted apoptosis in human cytomegalovirus-infected cells.
abstract: Mitochondria often play central roles in apoptotic pathways, and disruption of the mitochondrial transmembrane potential (ΔΨm) has been observed in various cells undergoing apoptosis. Human cytomegalovirus (HCMV) infection induces apoptosis in permissive cells; however, investigations of mitochondria-targeted apoptosis in HCMV-infected human foreskin fibroblast (HFF) cells have been limited. Here, we investigated the mitochondrial apoptosis pathway in HCMV-infected HFF cells. Flow cytometry analysis using JC-1 revealed that HCMV infection induces disruption of ΔΨm in HFF cells when administered 24 h postinfection (hpi), and this disruption was maximized at 48 hpi. Moreover, cytochrome c, normally a mitochondrial inner membrane protein, was detected in cytoplasmic extracts of HCMV-infected cells, but not mock-infected cells, by western blot analysis at 24 hpi. A caspase activity assay based on fluorescence spectrophotometry using a fluorogenic substrate revealed an increase in caspase-3 activity at 48 hpi in HCMV-infected cells. Caspase-8 activity was increased at 72 hpi in HCMV-infected cells. These results imply that HCMV infection induces mitochondria-mediated apoptosis in HFF cells.
authors.affiliation: Department of Microbiology, College of Natural Sciences, Chungbuk National University, Cheongju 361-763, Republic of Korea.
authors.forename: Gyu-Cheol
authors.initials: GC
authors.lastname: Lee
authors.forename: Jae Ho
authors.initials: JH
authors.lastname: Lee
authors.forename: Bo Yeon
authors.initials: BY
authors.lastname: Kim
authors.forename: Chan Hee
authors.initials: CH
authors.lastname: Lee
mesh_terms: D017209:Apoptosis; D015153:Blotting, Western; D053181:Caspase 8 / Q000032:analysis; D002478:Cells, Cultured; D045304:Cytochromes c / Q000032:analysis; D003587:Cytomegalovirus / Q000502:physiology*; D003593:Cytoplasm / Q000737:chemistry; D005347:Fibroblasts / Q000502:physiology / Q000821:virology*; D005434:Flow Cytometry; D006801:Humans; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria / Q000378:metabolism* / Q000502:physiology; D013050:Spectrometry, Fluorescence
chemical_list: D045304:Cytochromes c; D053181:Caspase 8