Crosslinking of proteins to ribosomal RNA in HeLa cell polysomes by sodium periodate. |
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Authors: | A J Svobada E H McConkey |
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Affiliation: | Department of Molecular, Cellular, and Developmental Biology University of Colorado Boulder, Colorado 80309 USA |
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Abstract: | HeLa cell polysomes were oxidized with sodium periodate and reduced with sodium borohydride to induce covalent crosslinks between ribosomal RNA and nearby proteins. We proved that RNA was tryly crosslinked to protein in oxidized, and not in control, samples using denaturing cesium trichloroacetate density gradients and phenol extraction. By both one- and two-dimensional gel analysis, we found that protein S3a can be crosslinked to 18S RNA, protein L3 to 28S RNA, and proteins L7′ and L23′ to 5.8S RNA. Because of the specificity of the periodate reaction, and since we were able to crosslink protein S1 to 16S RNA in , 30S ribosomal subunits, it is likely that we have crosslinked proteins to the 3′OH ends of HeLa polysomal RNAs. |
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Keywords: | SDS sodium dodecyl sulfate CsTCA cesium trichloroacetate TCA trichloroacetic acid BSA bovine serum albumen |
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