Structures of complexes of 5S RNA with ribosomal proteins L5, L18 and L25 from Escherichia coli: Identification of kethoxal-reactive sites on the 5S RNA

Abstract

The topography of Escherichia coli 5S RNA has been examined in the presence of ribosomal proteins L5, L18 and L25 and their different combinations, by comparing the kethoxal modification characteristics of the various RNA-protein complexes with those of the free A-conformer of 5S RNA (Noller & Garrett, 1979, accompanying paper). Two of the four most reactive guanines, G 13 and G 41, are unaffected by the protein, in accord with the finding that these are the only two guanines that are accessible in the 50S subunit (Noller & Herr, 1974). The other two very reactive guanines, G 24 and G 69, are strongly protected by protein L18, either in the presence or absence of proteins L5 and L25. Protein binding studies with kethoxal-modified 5S RNA provide evidence that one or both of these two guanines are directly involved in the protein-RNA interactions, and this conclusion is supported by the occurrence of guanines in these two positions in all the other sequenced prokaryotic 5S RNAs. The group of less reactive guanines, G 16, G 23, G 44, G 86 and G 107, are protected to some extent by each of the proteins L5, L18 and L25; the strongest effect is with L18. We suggest that this is attributable to a small increase in the conformational homogeneity of the 5S RNA and that L18, in particular, induces some tightening of the RNA structure. Only one guanine, G 69, is rendered more accessible by the proteins. This effect is produced by protein L25, which is known to cause some destructuring of the 5S RNA (Bear et al., 1977). There was no other evidence for any destructuring of the 5S RNA. In particular, the sequence 72 to 83, which is complementary to a sequence in 23S RNA (Herr & Noller, 1975), is not modified. However, in contrast to an earlier report (Erdmann et al., 1973), the conserved sequence G 44-A-A-C, which has been implicated in tRNA binding, was not rendered more accessible by the proteins.