Polypeptide chain initiation in eukaryotes: initiation factor requirements for translation of natural messengers

A protein-synthesizing system consisting of ribosomes and supernatant of undeveloped Artemia saline embryos is used for assay of mRNA translation and initiation factors. This system contains the components needed for chain elongation but has low levels of mRNA and initiation factors. Exogenous mRNA is readily translated upon addition of high-speed supernatant or ribosomal salt wash of developing embryos as a source of initiation factors (IF). This requirement can be largely satisfied by a mixture of the reticulocyte factors IF-MP, IF-M3, IF-M2A, and IF-M2B. A. salina IF-M1, which is present in undeveloped embryo supernatant, can be inactivated by A. salina IF-M1 antibody without affecting translation.