Abstract
Ribosomal subunits from the chloroplasts of Alaskan peas have been studied by immunoelectronmicroscopy. Electron micrographs of negatively stained small and large ribosomal subunits show particles of similar size and in the same characteristic projections described for the ribosomal subunits of Escherichia coli (Lake, J. A. (1976) J. Mol. Biol. 105, 131-159), although minor structural differences are apparent. High pressure liquid chromatographic analysis shows the modified nucleoside N6,N6-dimethyladenosine is conserved in chloroplast 16 S ribosomal RNA, presumably as two successive residues near the 3' end. Antibodies directed against N6,N6-dimethyladenosine were allowed to react with chloroplast 30S ribosomal subunits. Electron microscopy showed individual subunit-antibody complexes and pairs of ribosomal subunits cross-linked by a single antibody. In 94% of the complexes observed, antibody contact was consistent with a dimethyladenosine localization near the end of the small subunit platform, in an area of subunit contact in the 70 S ribosome. This localization is analogous to the placement of N6,N6-dimethyladenosine in the E. coli ribosome (Politz, S. M., and Glitz, D. G. (1977) Proc. Natl. Acad. Sci. U. S. A. 74, 1468-1472).
Highlights
Ribosomal subunits from the chloroplasts of Alasktaunres
Electron micrographs of negatively stained smalland volved in puromycin binding ( 7 ),and to investigate the struclarge ribosomal subunits show particleosf similar size ture of ribosomal RNA [8,9,10]
With regard to the E. coli ribosome, much is known about acetate, 6 mM 2-mercaptoethanol, 40 mM Tricine, pH 7.6) and centrifuged for 3 h at 200,OOO X g (55,000 rprn in a Beckman 60 Ti rotor)
Summary
ELECTRON MICROSCOPY OF RIBOSOMAL SUBUNITS AND LOCALIZATIONOF N6,N6DIMETHYLADENOSINE BY IMMUNOELECTRONMICROSCOPY*. Ribosomal subunits from the chloroplasts of Alasktaunres. It has been used to map the distribution of ribosomal peas have been studied ibmymunoelectronmicroscopy. Proteins [5, 6], to localize the ribosomal neighborhood in-. Electron micrographs of negatively stained smalland volved in puromycin binding ( 7 ) ,and to investigate the struclarge ribosomal subunits show particleosf similar size ture of ribosomal RNA [8,9,10]. Rather little is known about the detailed strucfor the ribosomal subunits Eosfcherichia coli(Lake, J. ture of chloroplast ribosomes. MOL BioL 105,131-159),althoughminor microscopy in studies of bacterial and rat liver cytoplasmic structural differences are apparent. Antibody contact was consistent with a dimethyladenosine localization near the end of the small subunit
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