A rapid method for mapping exposed cytosines in polyribonucleotides. Application to tRNATrp (yeast, beef liver).

Molecular biology reports ·Vol. 6 ·No. 2 ·1980-07-31 ·Pages 83-7

Mashkova TD, Mazo AM, Scheinker VS, Beresten SF, Bogdanova SL, Avdonina TA, Kisselev LL

Abstract

A rapid method for mapping exposed cytosine residues in 5'-[32P]-labeled RNA molecules is suggested. The exposed cytosines (C's) are converted into uracyls (U's) by bisulphite treatment at pH 5.8 in the presence of Mg2+, followed by complete modification of the residual (non-exposed) C's by a methoxyamine and bisulphite mixture at pH 5.0. The control RNA is modified only by methoxyamine and bisulphite without the preliminary C leads to U conversion. The location of the exposed C's is determined by comparing the products of partial T1, T2, A and U2 ribonuclease digestions of the C leads to U converted and control RNAs after slab gel polyacrylamide electrophoresis and autoradiography. The method has been applied for mapping exposed cytosine bases in tRNATrp (yeast) which have been found in the anti-codon loop and at the 3'-end of the molecule. In tRNATrp (beef liver), in addition to the same exposed bases, C in the diHU-loop is exposed. The data obtained are in full agreement with what is known about exposed C's for other tRNAs.

MeSH Terms

Animals Autoradiography Base Sequence Cattle Cytosine/analysis Hydroxylamines RNA, Fungal/analysis RNA, Transfer/analysis Ribonucleases Saccharomyces cerevisiae/analysis Sulfites Tryptophan Uracil/analysis

Chemicals

Hydroxylamines RNA, Fungal Sulfites Uracil Tryptophan Cytosine RNA, Transfer Ribonucleases sodium bisulfite

Authors & Affiliations

7 authors, click to expand affiliations / ORCID

Mashkova T D

Mazo A M

Scheinker V S

Beresten S F

Bogdanova S L

Avdonina T A

Kisselev L L

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27 references, click to expand

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Article Info

Journal

Molecular biology reports

Abbr.

Mol Biol Rep

ISSN

0301-4851

DOI

10.1007/BF00778434

Published

1980-07-31

Pages

83-7

Language

English

Region

Netherlands

NLM ID

0403234

Subset

External Links

PubMed source DOI: 10.1007/BF00778434 View journal details

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A rapid method for mapping exposed cytosines in polyribonucleotides. Application to tRNATrp (yeast, beef liver).

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