Relationship of eukaryotic initiation factor 3 to poliovirus-induced p220 cleavage activity.
The cleavage of the p220 subunit of eukaryotic initiation factor 4F (eIF-4F) that is induced by the poliovirus protease 2A has been shown previously to require another translation initiation factor, eIF-3. The role of eIF-3 in this cleavage reaction, however, is not known. An antiserum was raised against human eIF-3 and used to analyze the eIF-3 subunit composition in poliovirus-infected and uninfected HeLa cells and after incubation of eIF-3 in vitro with viral 2A protease. No evidence for 2Apro-dependent cleavage of any eIF-3 subunit was detected. Infected cells contain an activity that catalyzes the cleavage of p220 to a specific set of cleavage products. This activity is thought to be an activated form of a latent cellular protease. The p220-specific cleavage activity was partially purified. It was resolved from eIF-3 by both gel filtration and anion-exchange chromatography. Neither intact eIF-3 nor any detectable subunits of eIF-3 were found to copurify with the p220-specific cleavage activity. The latter activity behaves as a protein of 55,000 to 60,000 molecular weight and is inhibited by alkylating agents and metals, which indicates the presence of essential thiol groups. When this activity was incubated with partially purified p220, cleavage occurred only in the presence of eIF-3. Thus, eIF-3 appears to play a role in the p220 cleavage cascade which is subsequent to the 2Apro-induced activation of the p220-specific protease.
Wyckoff
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Department of Cellular, Viral, and Molecular Biology, University of Utah School of Medicine, Salt Lake City 84132.
Lloyd
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Ehrenfeld
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D002457:Cell Extracts; D003546:Cysteine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D004789:Enzyme Activation; D039621:Eukaryotic Initiation Factor-3; D006367:HeLa Cells; D006801:Humans; D046911:Macromolecular Substances; D008954:Models, Biological; D010448:Peptide Initiation Factors / Q000302:isolation & purification / Q000378:metabolism*; D011051:Poliomyelitis / Q000378:metabolism*; D017955:Poliovirus / Q000201:enzymology*; D011480:Protease Inhibitors; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014764:Viral Proteins
D002457:Cell Extracts; D039621:Eukaryotic Initiation Factor-3; D046911:Macromolecular Substances; D010448:Peptide Initiation Factors; D011480:Protease Inhibitors; D013438:Sulfhydryl Compounds; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; C057400:picornain 2A, Picornavirus