Summary
Protein S8 occurred in two conformations. A minor component, constituting up to about 30% of the total protein, which bound to 16S RNA at 0°, and a major component which did not bind to the 16S RNA at 0°. The latter could be converted to a binding conformation by heating (E≊12 kcal/mole). The RNA binding site, for most of the RNA population, was stable at 0°, but a minimum Mg++ concentration of 5×10-3 M, and an optimum concentration of 3×10-2 M or higher, was necessary for this stability. Specific binding of protein S8 to 16S RNA occurred over a wide range of K+ concentrations, but below 0.25 M KCl there was some additional non-specific binding, and above 0.35 M KCl there was a gradual decrease in the binding affinity of S8 for 16S RNA. A maximum in the pH dependence binding curve was at pH 7.4 to 8.0.
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References
Cammack, K., Miller, D. S., Grinstead, K. H.: Physical properties of ribosomal RNA isolated from bacteria deficient in ribonuclease I. Biochem. J. 117, 745–755 (1970).
Fellner, P., Ehresmann, C., Stiegler, P., Ebel, J.-P.: The partial nucleotide sequence of the 16S ribosomal RNA from Escherichia coli. Nature (Lond.) New Biol. 239, 1–4 (1972).
Garrett, R. A., Rak, K. H., Daya, L., Stöffler, G.: Specific protein binding sites on 16S RNA of E. coli. Molec. gen. Genet. 114, 112–124 (1971).
Kaltschmidt, E., Wittmann, H. G.: Ribosomal proteins VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Analyt. Biochem. 36, 401–412 (1970).
Leboy, P. S., Cox, E. C., Flaks, J. G.: The chromosomal site specifying a ribosomal protein in Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 52, 1367–1374 (1964).
Mizushima, S., Nomura, M.: Assembly mapping of 30S ribosomal proteins from E. coli. Nature (Lond.) 226, 1214–1218 (1970).
Ohlenbusch, H. H., Olivera, B. M., Tuan, D., Davidson, N.: Selective dissociation of histones from calf thymus nucleoprotein. J. molec. Biol. 25, 299–315 (1967).
Schaup, H. W., Green, M., Kurland, C. G.: Identification of RNA binding sites for individual 30S ribosomal proteins. Molec. gen. Genet. 109, 193–205 (1970).
Schaup, H. W., Green, M., Kurland, C. G.: Molecular interactions of ribosomal components II. Site specific complex formation between 30S proteins and ribosomal RNA. Molec. gen. Genet. 112, 1–8 (1971).
Traub, P., Nomura, M.: Mechanism of assembly of 30S ribosomes studied in vitro. J. molec. Biol. 40, 391–413 (1969).
Wittmann-Liebold, B.: Amino acid composition of the tryptic peptides isolated from proteins S4, S18 and S20 of E. coli ribosomes. Hoppe-Seylers Z. physiol. Chem. 352, 1705–1714 (1971).
Zimmermann, R. A., Muto, A., Fellner, P., Ehresmann, C., Branlant, C.: Localisation of ribosomal protein binding sites on 16S ribosomal RNA. Proc. nat. Acad. Sci. (Wash.) 69, 1282–1286 (1972).
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Communicated by E. Bautz
Paper No. 41 on “Ribosomal Proteins”. Preceding paper is in press in Mol. Gen. Genetics (1972).
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Schulte, C., Garrett, R.A. Optimal conditions for the interaction of ribosomal protein S8 and 16S RNA and studies on the reaction mechanism. Molec. Gen. Genet. 119, 345–355 (1972). https://doi.org/10.1007/BF00272092
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DOI: https://doi.org/10.1007/BF00272092