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  • Title: Polypeptide chain initiation in eukaryotes: mechanism of formation of initiation complex.
    Author: Nombela C, Nombela NA, Ochoa S, Safer B, Anderson WF, Merrick WC.
    Journal: Proc Natl Acad Sci U S A; 1976 Feb; 73(2):298-301. PubMed ID: 1061132.
    Abstract:
    Artemia salina ribosomal subunits and highly purified reticulocyte initiation factors (IF) are used to study the mechanism of formation of the puromycin-sensitive initiation complex Met-tRNAi-80S ribosome-AUG. A complex with equimolar amounts of 40S subunit, GTP, and Met-tRNAi is formed at low Mg2+ concentration with a requirement for IF-MP (homogeneous) but not AUG or other factors. An 80S complex is formed only upon the further addition of AUG, IF-M2A, and IF-M2B, but not of either factor alone. This complex contains no GTP or GDP. A 40S complex, which cannot be converted to an 80S one, is formed when the nonhydrolyzable analog GMPPCP is substituted for GTP. IF-M2A has no effect on the formation of this complex, but IF-M2B enhances its formation.
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