Accessibility of guanine at position 44 in the invariant sequence 5'CCG44AAC3' of Escherichia coli 5S RNA to reaction with kethoxal.

1 September 1979

journal article
research article
Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences

Vol. 76 (9) , 4400-4404
https://doi.org/10.1073/pnas.76.9.4400

Abstract

The reaction of E. coli ribosomes with .beta.-ethoxy-.alpha.-ketobutyraldehyde (kethoxal) in a buffer containing 50-100 mM tris .cntdot. HCl at pH 7.4, 50 mM NH4Cl, and 5 mM Mg(OAc)2 readily released the 5S RNA from the ribosomes. When liberated, the 5S RNA is in a conformation such that position 44 is selectively reactive, in addition to the normally reactive guanines at positions 41 and 13. Positions 41 and 13 were previoiusly shown to react in the 5S RNA in situ. The resulting new RNase Tl resistant oligonucleotides .**GRAPHIC**. and .**GRAPHIC**. were isolated and identified. These oligonucelotides have not been found in RNase Tl digests of 5S RNA that is not released from the ribosome. The guanine at position 44 is part of the invariant sequence 5''CCG44AAC3''which includes the portion of the molecule thought to interact with the invariant 5''-GT.psi.C3'' of tRNA in the ribosomal A site. This invariant sequence of the 5S RNA may also form part of the binding site for protein L5.

Keywords

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