Wheat germ cytoplasmic ribosomes. Structure of ribosomal subunits and localization of N6,N6-dimethyladenosine by immunoelectron microscopy.
Cytoplasmic ribosomes have been isolated from wheat germ, and the structure of ribosomal subunits has been examined by electron microscopy of negatively stained preparations. Small (40 S) subunits show structural features generally regarded as characteristic of eukaryotic particles, while large (60 S) subunits show shapes that are equally well described by models of prokaryotic 50 S particles. Small subunit 18 S RNA contains 2 residues of N6,N6-dimethyladenosine 19 and 20 residues from the 3'-end (Hagenbüchle, O., Santer, M., Steitz, J. A., and Mans, R. J. (1978) Cell 13, 551-563). Nucleoside analysis by high performance liquid chromatography shows no other residues of this component in the RNA. Anti-dimethyladenosine immunoglobulins were reacted with wheat germ 40 S subunits, and the resulting complexes were studied by electron microscopy in order to localize the nucleoside. In about 90% of the complexes observed, antibody-subunit contact was consistent with a single binding site. We place the dimethyladenosine residues at or near the end of the platform of the 40 S particle in a position nearly equivalent to that previously identified in prokaryotic and chloroplast subunits (Trempe, M. R., and Glitz, D. G. (1981) J. Biol. Chem. 256, 11873-11879).
Montesano
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Department of Biological Chemistry, School of Medicine, University of California, Los Angeles 90024.
Glitz
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DG
D000241:Adenosine / Q000031:analogs & derivatives* / Q000032:analysis; D003593:Cytoplasm / Q000648:ultrastructure; D007118:Immunoassay; D008854:Microscopy, Electron; D010944:Plants / Q000648:ultrastructure*; D012315:RNA Caps / Q000032:analysis*; D012337:RNA, Ribosomal, 18S / Q000032:analysis; D012270:Ribosomes / Q000648:ultrastructure*; D014908:Triticum / Q000648:ultrastructure
D012315:RNA Caps; D012337:RNA, Ribosomal, 18S; C021013:N(6),N(6)-dimethyladenosine; D000241:Adenosine