title: The RIP-like kinase, RIP3, induces apoptosis and NF-kappaB nuclear translocation and localizes to mitochondria.
abstract: A RIP-like protein, RIP3, has recently been reported that contains an N-terminal kinase domain and a novel C-terminal domain that promotes apoptosis. These experiments further characterize RIP3-mediated apoptosis and NF-kappaB activation. Northern blots indicate that rip3 mRNA displays a restricted pattern of expression including regions of the adult central nervous system. The rip3 gene was localized by fluorescent in situ hybridization to human chromosome 14q11.2, a region frequently altered in several types of neoplasia. RIP3-mediated apoptosis was inhibited by Bcl-2, Bcl-x(L), dominant-negative FADD, as well as the general caspase inhibitor Z-VAD. Further dissection of caspase involvement in RIP3-induced apoptosis indicated inhibition by the more specific inhibitors Z-DEVD (caspase-3, -6, -7, -8, and -10) and Z-VDVAD (caspase-2). However, caspase-1, -6, -8 and -9 inhibitors had little or no effect on RIP3-mediated apoptosis. Mutational analysis of RIP3 revealed that the C-terminus of RIP3 contributed to its apoptotic activity. This region is similar, but distinct, to the death domain found in many pro-apoptotic receptors and adapter proteins, including FAS, FADD, TNFR1, and RIP. Furthermore, point mutations of RIP3 at amino acids conserved among death domains, abrogated its apoptotic activity. RIP3 was localized by immunofluorescence to the mitochondrion and may play a key role in the mitochondrial disruptions often associated with apoptosis.
authors.affiliation: AstraZeneca Pharmaceuticals, Enabling Science and Technology Department, 1800 Concord Pike, Wilmington, DE 19803, USA.
authors.forename: G M
authors.initials: GM
authors.lastname: Kasof
authors.forename: J C
authors.initials: JC
authors.lastname: Prosser
authors.forename: D
authors.initials: D
authors.lastname: Liu
authors.forename: M V
authors.initials: MV
authors.lastname: Lorenzi
authors.forename: B C
authors.initials: BC
authors.lastname: Gomes
mesh_terms: D000595:Amino Acid Sequence; D017209:Apoptosis / Q000502:physiology*; D015152:Blotting, Northern; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002883:Chromosomes, Human, Pair 14; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008928:Mitochondria / Q000737:chemistry*; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
chemical_list: D061945:Caspase Inhibitors; D016328:NF-kappa B; D011494:Protein Kinases; C119705:RIPK3 protein, human; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; D020169:Caspases