title: Structural determinants of caspase-9 inhibition by the vaccinia virus protein, F1L.
abstract: In multicellular organisms, apoptosis is a powerful method of host defense against viral infection. Apoptosis is mediated by a cascade of caspase-family proteases that commit infected cells to a form of programmed cell death. Therefore, to replicate within host cells, viruses have developed various strategies to inhibit caspase activation. In the mitochondrial cell-death pathway, release of cytochrome c from mitochondria into the cytosol triggers assembly of the oligomeric apoptosome, resulting in dimerization and activation of the apical caspase-9 (C9), and in turn its downstream effector caspases, leading to apoptosis. We previously showed that the vaccinia virus-encoded Bcl-2-like protein, F1L, which suppresses cytochrome c release by binding Bcl-2 family proteins, is also a C9 inhibitor. Here, we identify a novel motif within the flexible N-terminal region of F1L that is necessary and sufficient for interaction with and inhibition of C9. Based on functional studies and mutagenesis, we developed an atomic model of the complex in which F1L inhibits C9 by engaging the active site in the reverse orientation with respect to substrate peptides, in a manner analogous to that of XIAP-mediated inhibition of caspases-3 and -7. These studies offer new insights into the mechanism of apoptosome inhibition by F1L as well as novel probes to understand the molecular bases of apoptosome regulation and turnover. They also suggest how the two distinct functionalities of F1L (inhibition of C9 and suppression of pro-apoptotic Bcl-2 family proteins) may operate in a cellular setting.
authors.affiliation: Sanford-Burnham Medical Research Institute, La Jolla, California 92037.
authors.forename: Eric
authors.initials: E
authors.lastname: Yu
authors.affiliation: Sanford-Burnham Medical Research Institute, La Jolla, California 92037.
authors.forename: Dayong
authors.initials: D
authors.lastname: Zhai
authors.affiliation: Sanford-Burnham Medical Research Institute, La Jolla, California 92037.
authors.forename: Chaofang
authors.initials: C
authors.lastname: Jin
authors.affiliation: Sanford-Burnham Medical Research Institute, La Jolla, California 92037.
authors.forename: Motti
authors.initials: M
authors.lastname: Gerlic
authors.affiliation: Sanford-Burnham Medical Research Institute, La Jolla, California 92037.
authors.forename: John C
authors.initials: JC
authors.lastname: Reed
authors.affiliation: Sanford-Burnham Medical Research Institute, La Jolla, California 92037. Electronic address: rlidding@sanfordburnham.org.
authors.forename: Robert
authors.initials: R
authors.lastname: Liddington
mesh_terms: D000595:Amino Acid Sequence; D017209:Apoptosis; D053453:Caspase 9 / Q000378:metabolism*; D016923:Cell Death; D057809:HEK293 Cells; D006801:Humans; D007113:Immunity, Innate; D008954:Models, Biological; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D014616:Vaccinia virus / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: C507356:F1L protein, vaccinia virus; D010455:Peptides; D014764:Viral Proteins; D053453:Caspase 9