Binding of Met-tRNAf to native 40 S ribosomal subunits in Ehrlich ascites tumor cells

Abstract

Two forms of native 40 S ribosomal subunits, distinguishable by their buoyant densities, are recovered from Ehrlich ascites cells. In this communication, we describe experiments designed to test whether Met-tRNAf is associated with either form. Our results indicate that (a) in the cell, Met-tRNAf is bound to the native 40 S subunit, and in particular, to the subunit of density 1.40 g x cm-3; (b) under the growth conditions used, less than 10% of the low density native subunits have bound Met-tRNAf; (c) the majority of the Met-tRNAf containing native 40 S subunits, isolated from sucrose gradient analyses of cell extracts, join with 60 S subunits in vitro to form 80 S monosomes only if additional ribosomal wash factors are provided; and (d) the 80 S monosomes so formed are completed initiation complexes that can form peptide bonds. These results support the hypothesis that various forms of the native 40 S subunit represent different stages in the formation of a complete initiation complex.