Structure of 18 S ribosomal RNA in native 40 S ribosomal subunits

Abstract

We have analyzed the structure of 18 S rRNA in native 40 S subunits using chemical modification followed by primer extension. The native subunits were modified using the single-stranded specific reagents dimethyl sulfate and 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide metho-p-toluenesulfonate. The modification pattern of the 18 S rRNA was compared to that obtained from derived 40 S subunits prepared by dissociation of unprogrammed 80 S ribosomes. Eighteen nucleotides showed different accessibility to the chemical probes in derived and native subunits. Half of these nucleotides were found in the central domain of the rRNA between the 1060 loop and the central pseudoknot. The remaining nucleotides were located in two clusters in the 5'- and 3'-domains of the 18 S rRNA. Derived 40 S subunits are free from non-ribosomal proteins. In contrast, native subunits are intermediates in protein synthesis initiation and contain stoichiometric amounts of initiation factor 3 (Sundkvist, I. C., and Staehelin, T. (1975) J. Mol. Biol. 99, 401- 418). The possible role of this factor in altering the structure of 18 S rRNA in the native 40 S subunits is discussed.