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

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→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 → 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 anticodon 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 that is known about exposed C's for other tRNAs.

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References

1. Rich A. & RajBhandary U.L. Ann. Rev. Biochem. 45, 805–858 (1976).
   Google Scholar
2. Goddard J.P., Progr. Biophys. Mol. Biol. 32, 233–308 (1977).
   Google Scholar
3. Kim, S.-H., In: Transfer RNA (Altman, S., ed.), The MIT Press, pp. 248–293 (1978).
4. Jilyaeva T. & Kisselev L. FEBS Letters 10, 229–232 (1970).
   Google Scholar
5. Kucan Z., Freude K.A., Kucan I. & Chambers R.W. Nature New Biol. 232, 177–180 (1971).
   Google Scholar
6. Singhal R.P. J. Biol. Chem. 43, 245–250 (1971).
   Google Scholar
7. Goddard J.P. & Schulman L.H. J. Biol. Chem. 247, 3864–3867 (1972).
   Google Scholar
8. Schmidt F.J., Omilianowski D.R. & Bock R.M. Biochemistry 12, 4980–4983 (1973).
   Google Scholar
9. Chambers R.W., Aoyagi Sh., Firukawa Y., Zawadska H. & Chanot O.S. J. Biol. Chem. 248, 5549–5551 (1973).
   Google Scholar
10. Chakraburtty K. Nucleic Acids Res. 2, 1793–1804 (1975).
    Google Scholar
11. Batey I.L. & Brown D.M. Mol. Biol. Rep. 2, 65–72 (1975).
    Google Scholar
12. Batey I.L. & Brown D.M. Biochim. Biophys. Acta 474, 378–385 (1977).
    Google Scholar
13. Bhanot O.S. & Chambers R.W. J. Biol. Chem. 252, 2551–2559 (1977).
    Google Scholar
14. Schulman L.M. & Pelka H. Biochem. 16, 4256–4265 (1977).
    Google Scholar
15. Hayatsu H. In: Progress in Nucleic Acids Research and Molecular Biology (Cohn W.E., ed.), Academic Press, N.-Y., 16, pp. 75–120 (1976).
    Google Scholar
16. Sverdlov E.D., Monastyrskaya G.S., Guskova L.L., Levitan T.L., Scheichenko V.I. & Budovsky E.I. Biochim. Biophys. Acta 340, 153–165 (1974).
    Google Scholar
17. Donis-Keller H., Maxam A.M. & Gilbert W. Nucleic Acids Res. 4, 2527–2538 (1977).
    Google Scholar
18. Vlassov V.V., Tchizhikov V.E., Scheinker V.Sh. & Favorova O.O. FEBS Letters 90, 103–106 (1978).
    Google Scholar
19. Richardson C.C. Proc. Nat. Acad. Sci. USA 54, 158–165 (1965).
    Google Scholar
20. Mazo A.M., Scheinker V.S. & Kisselev L.L. Mol. Biol. Rep. 2, 233–239 (1975).
    Google Scholar
21. Simonesits A., Brownlee G.G., Brown R.S., Rubin J.R. & Guilley H. Nature 269, 833–836 (1977).
    Google Scholar
22. Stanley J. & Vassilenko S. Nature 274, 87–89 (1978).
    Google Scholar
23. Peattie D.A. Proc. Nat. Acad. Sci. USA 76, 1760–1764 (1974).
    Google Scholar
24. Mazo A.M., Mashkova T.D., Avdonina T.A., Ambartsumyan N.S. & Kisselev L.L. Nucleic Acids Res. 7, 2469–2482 (1979).
    Google Scholar
25. Mazo A.M. & Kisselev L.L. FEBS Letters 59, 177–179 (1975).
    Google Scholar
26. Mazo A.M., Avdonina T.A., Mashkova T.D. & Kisselev L.L. Biochim. Biophys. Acta 432, 353–360 (1976).
    Google Scholar
27. Rushizky G.W., Mozeiko J.H., Rogerson D.L. & Sober H.A. Biochemistry 9, 4966–4971 (1970).
    Google Scholar
28. Krupp, G. & Gross, J.H. Nucleic Acids Res. (1979) in press.
29. Keith G., Roy A., Ebel J.-P. & Dirheimer G. Biochimie 54, 1405–1426 (1972).
    Google Scholar
30. Fournier M., Labouesse J., Dirheimer G., Fix C. & Keith G. Biochim, Biophys. Acta 521, 198–208 (1978).
    Google Scholar

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Authors and Affiliations

1. Institute of Molecular Biology, the USSR Academy of Sciences, V-334, Moscow, USSR
   T. D. Mashkova, A. M. Mazo, V. Sh. Scheinker, S. F. Beresten, S. L. Bogdanova, T. A. Avdonina & L. L. Kisselev

Authors

1. T. D. Mashkova
2. A. M. Mazo
3. V. Sh. Scheinker
4. S. F. Beresten
5. S. L. Bogdanova
6. T. A. Avdonina
7. L. L. Kisselev

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Mashkova, T.D., Mazo, A.M., Scheinker, V.S. et al. A rapid method for mapping exposed cytosines in polyribonucleotides. Application to tRNATrp (yeast, beef liver).Mol Biol Rep 6, 83–87 (1980). https://doi.org/10.1007/BF00778434

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• Received: 23 October 1979
• Issue date: July 1980
• DOI: https://doi.org/10.1007/BF00778434

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