Formation of a 22S mRNA X rRNA X protein complex during translation of globin messenger RNA

Abstract

Poly(A)-rich RNA was isolated from rabbit reticulocyte polyribosomes by affinity chromatography on oligo(dT)-cellulose and fractionated in sucrose gradients under non-denaturing conditions. Most of the translatable RNA sedimented in sucrose gradients both as free 9S mRNA and as a 22S complex containing 18S ribosomal RNA and a protein of Mr 66 000. The complex was characterized by identification of the translation products. Experiments with both labelled globin mRNA and Mr-66 000 protein indicate that the complex is not an artefact, but rather that it is formed during the initiation of protein synthesis. The Mr-66 000 protein appears to be a component of the 48S pre-initiation complex and recycles before 80S complex formation.