Conservation of structure and function in vertebrate c-FLIP proteins despite rapid evolutionary change
| Autor: | Kazuhiro Sakamaki, Chisa Shukunami, Kentaro Tomii, Naoto Ueno, Hiroaki Iwata, Kumiko Chiba, Chiyo Takagi, Kenichiro Imai, Naoyuki Iwabe |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Subfamily
RT-PCR reverse transcription-polymerase chain reaction Evolution Mutant Biophysics Xenopus Apoptosis Biochemistry NF-κB CFLAR CASc Caspase interleukin-1 β converting enzyme homologs tubα6 tubulin α6 Protein structure PCR polymerase chain reaction CHX cycloheximide biology.animal CARD caspase-recruitment domain DED death effector domain MO morpholino oligonucleotide Zebrafish TRAF2 tumor necrosis factor receptor-associated factor 2 Pseudocatalytic triad Genetics Protein subfamily biology Vertebrate NF-κB Nuclear factor-kappa B biology.organism_classification FADD Fas-associated death domain protein c-FLIP cellular FLICE-like inhibitory protein EGFP enhanced green fluorescent protein ODC ornithine decarboxylase Embryogenesis Caspase-8 Research Article |
| Zdroj: | Biochemistry and Biophysics Reports |
| ISSN: | 2405-5808 |
| Popis: | Cellular FLICE-like inhibitory protein (c-FLIP, gene symbol CFLAR) was first identified as a negative regulator of death receptor-mediated apoptosis in mammals. To understand the ubiquity and diversity of the c-FLIP protein subfamily during evolution, c-FLIP orthologs were identified from a comprehensive range of vertebrates, including birds, amphibians, and fish, and were characterized by combining experimental and computational analysis. Predictions of three-dimensional protein structures and molecular phylogenetic analysis indicated that the conserved structural features of c-FLIP proteins are all derived from an ancestral caspase-8, although they rapidly diverged from the subfamily consisting of caspases-8, -10, and -18. The functional role of the c-FLIP subfamily members is nearly ubiquitous throughout vertebrates. Exogenous expression of non-mammalian c-FLIP proteins in cultured mammalian cells suppressed death receptor-mediated apoptosis, implying that all of these proteins possess anti-apoptotic activity. Furthermore, non-mammalian c-FLIP proteins induced NF-κB activation much like their mammalian counterparts. The CFLAR mRNAs were synthesized during frog and fish embryogenesis. Overexpression of a truncated mutant of c-FLIP in the Xenopus laevis embryos by mRNA microinjection caused thorax edema and abnormal constriction of the abdomen. Depletion of cflar transcripts in zebrafish resulted in developmental abnormalities accompanied by edema and irregular red blood cell flow. Thus, our results demonstrate that c-FLIP/CFLAR is conserved in both protein structure and function in several vertebrate species, and suggest a significant role of c-FLIP in embryonic development. Highlights • c-FLIP/CFLAR is specific to the vertebrate lineage. • A rapid evolutionary divergence of the c-FLIP subfamily members has occurred. • The pseudocatalytic triad is formed by evolutionarily conserved amino acids. • Vertebrate c-FLIP proteins work on both apoptotic inhibition and NF-κB activation. • c-FLIP proteins play a developmental role during embryogenesis of frog and fish. |
| Databáze: | OpenAIRE |
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