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Homologies in processing and sequence between the 23S ribosomal ribonucleic acids of Paracoccus denitrificans and Rhodopseudomonas sphaeroides
Authors:R. M. MacKay  L. B. Zablen  C. R. Woese  W. F. Doolittle
Affiliation:(1) Department of Biochemistry, Tupper Medical Building Dalhousie University, B3H 4H7 Halifax, N.S., Canada;(2) Department of Genetics and Development, University of Illinois, 61801 Urbana, Ill., USA
Abstract:We have shown that mature 50S ribosomal subunits of Paracoccus denitrificans lack intact 23S rRNA, containing instead rRNAs of 0.56 (ldquo16Srdquo) and 0.37 (ldquo14Srdquo)x106 molecular weight. Kinetic labelling studies showed these to be derived from a 1.02x106 dalton precursor, which may itself derive from a larger and very transient ldquo23Srdquo species. A similar pattern of rRNA processing has been previously described for Rhodopseudomonas sphaeroides, and we have compared, by Tl oligonucleotide catalog analysis, the smaller (ldquo14Srdquo) fragments of P. denitrificans and R. sphaeroides 23S rRNAs. These were shown to exhibit strong sequence homology, and comparisons of 14S-derived oligonucleotides to oligonucleotides from an in vitro-generated ldquo13Srdquo fragment of Escherichia coli 23S rRNA suggest that P. denitrificans and R. sphaeroides 14S rRNAs arise from the 5prime-terminal portions of their respective 23S precursors. Results are considered to be consistent with the claim that P. denitrificans arose, by loss of photophosphorylation, from a member of the Rhodospirillaceae.Abbreviations E buffer 60 ml 2 M Tris base, 20 ml 3 M sodium acetate, 15 ml 0.2M disodium EDTA, 6 ml glacial acetic acid, 900 ml distilled water - HEPES N-2-hydroxymethyl piperazine-Nprime-2-ethanesulfonic acid - TMK 5 mM Tris-Cl, 0.1 mM MgSO4, 60 mM KCl. pH 7.3 - TM3 10 mM Tris-Cl, 1 mM MgCl2, pH 7.3 - Tris tris (hydroxymethyl) aminomethane - SDS sodium dodecyl sulphate
Keywords:Paracoccus denitrificans  Rhodopseudomonas sphaeroides  rRNA sequences  rRNA processing
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