Unusual requirements for optimum translation of polio viral RNA in vitro

• Published in: The Journal of biological chemistry 1983-06, Vol.258 (11), p.7195-7199
• Main Authors: Daniels-McQueen, S, Detjen, B M, Grifo, J A, Merrick, W C, Thach, R E
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Carcinoma, Krebs 2 - metabolism; Encephalomyocarditis virus - genetics; Fundamental and applied biological sciences. Psychology; HeLa Cells - physiology; Humans; Kinetics; Mice; Microbiology; Poliovirus - genetics; Protein Biosynthesis; Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains; Ribosomes - metabolism; RNA, Viral - genetics; Virion - genetics; Virology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)32351-2

The translation of poliovirion RNA (polio RNA) in an in vitro fractionated system was much less efficient than that of encephalomyocarditis virion RNA (EMC RNA). In contrast, when polio and EMC RNAs were added to postmitochondrial cell lysates (S10), they were translated with equal efficiency. However, this equality was observed only when high concentrations of S10 were employed; at lower concentrations, polio RNA translation was reduced relative to that of EMC RNA. These results suggest that both the fractionated and S10 systems are limiting in a component that is required for the optimal translation of polio RNA. The elongation rates for EMC and polio RNA translation in the fractionated system were found to be similar, indicating that this component acts at an initiation step. Various components, including excess ribosomal salt wash and postribosomal supernatant of cell lysate, were added to the fractionated system in an effort to identify the slow step more precisely. Of these, only excess ribosomal salt wash specifically stimulated polio RNA translation, suggesting that one or more initiation factors is necessary in unusually large amounts for this mRNA. Various purified initiation factors were tested for the ability to enhance polio RNA translation. Of these, only purified eukaryotic initiation factor 4A had a specific effect. This suggests that polio RNA, in contrast to other mRNAs tested (EMC, reoviral, and globin), may have an unusually low affinity for this initiation factor. The significance of these results is discussed in terms of the methods picornaviruses have evolved for reprogramming the translational machinery of the host cell.