Eukaryotic translation initiation factor 4G is targeted for proteolytic cleavage by caspase 3 during inhibition of translation in apoptotic cells.
Although much is known about the multiple mechanisms which induce apoptosis, comparatively little is understood concerning the execution phase of apoptosis and the mechanism(s) of cell killing. Several reports have demonstrated that cellular translation is shut off during apoptosis; however, details of the mechanism of translation inhibition are lacking. Translation initiation factor 4G (eIF4G) is a crucial protein required for binding cellular mRNA to ribosomes and is known to be cleaved as the central part of the mechanism of host translation shutoff exerted by several animal viruses. Treatment of HeLa cells with the apoptosis inducers cisplatin and etoposide resulted in cleavage of eIF4G, and the extent of its cleavage correlated with the onset and extent of observed inhibition of cellular translation. The eIF4G-specific cleavage activity could be measured in cell lysates in vitro and was inhibited by the caspase inhibitor Ac-DEVD-CHO at nanomolar concentrations. A combination of in vivo and in vitro inhibitor studies suggest the involvement of one or more caspases in the activation and execution of eIF4G cleavage. Furthermore recombinant human caspase 3 was expressed in bacteria, and when incubated with HeLa cell lysates, was shown to produce the same eIF4G cleavage products as those observed in apoptotic cells. In addition, purified caspase 3 caused cleavage of purified eIF4G, demonstrating that eIF4G could serve as a substrate for caspase 3. Taken together, these data suggest that cellular translation is specifically inhibited during apoptosis by a mechanism involving cleavage of eIF4G, an event dependent on caspase activity.
Marissen
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Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73190, USA.
Lloyd
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RE
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D002945:Cisplatin / Q000494:pharmacology; D003574:Cytochrome c Group / Q000494:pharmacology; D003838:Deoxyadenine Nucleotides / Q000494:pharmacology; D005047:Etoposide / Q000494:pharmacology; D039603:Eukaryotic Initiation Factor-4G; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D009842:Oligopeptides / Q000494:pharmacology; D010448:Peptide Initiation Factors / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D014176:Protein Biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism
D000590:Amino Acid Chloromethyl Ketones; D003574:Cytochrome c Group; D003838:Deoxyadenine Nucleotides; D039603:Eukaryotic Initiation Factor-4G; D009842:Oligopeptides; D010448:Peptide Initiation Factors; D011480:Protease Inhibitors; D011994:Recombinant Proteins; C099158:acetyl-aspartyl-glutamyl-valyl-aspartal; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D005047:Etoposide; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; C026600:2'-deoxyadenosine triphosphate; D002945:Cisplatin