Localization of the 3′ End of 16S rRNA in Escherichia coli 30S ribosomal subunits by Immuno electron microscopy

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Abstract
Summary The location of the 3′ end of 16S rRNA in E. coli 30S ribosomal subunits has been determined by immuno electron microscopy. The 3′ terminal adenosine of isolated 16S rRNA was oxidized with sodium periodate and reacted with N-γ-(2,4-dinitrophenyl) aminobutyric acid hydrazide. Functionally active 30S subunits were reconstituted from DNP-16S rRNA and total 30S ribosomal proteins. DNP-30S subunits were complexed with DNP-specific IgG-antibody and examined in the electron microscope. The 3′ end of the 16S rRNA was mapped at a single region located at the inner side of the large lobe of the 30S subunit. The location of the 3′ end also provides information as to the topography of the binding domain of natural mRNA on 30S subunits, since a pyrimidine-rich sequence at the 3′ terminal region of 16S rRNA participates in the correct alignment of natural mRNAs during initiation complex formation.