Inactivation of rat liver RNA polymerases I and II and yeast RNA polymerase I by pyrodixal 5'-phosphate. Evidence for the participation of lysyl residues at the active site.
[en] Purified DNA-dependent RNA polymerase forms I (A) and II (B) from rat liver and form I from yeast are rapidly inactivated by pyridoxal 5'-phosphate at pH 8.0. The inhibition is relatively specific since pyridoxamine 5'-phosphate is not an inhibitor and pyridoxal is about 12 times less effective than pyridoxal 5'-phosphate. The inactivation is reversed by high concentrations of amines, and can be made irreversible by reduction with NaBH4. Spectral analysis of the inhibited enzyme and its NaBH4 reduction product indicates that a Schiff base forms between the aldehyde group of pyridoxal 5'-phosphate and one or more amino groups of the protein. Nepsilon-Pyridoxyllysine was identified as the only product in acid hydrolysates of the reduced yeast RNA polymerase I-pyridoxal 5'-phosphate complex. Complete inactivation of yeast polymerase I results in the incorporation of 3-4 mol of pyridoxal 5'-phosphate/1 mol of enzyme. DNA and nucleotide substrates partially protect the enzymes from inactivation. These results suggest that one or more lysyl amino groups are critical for the activity of animal RNA polymerases and show that pyridoxal 5'-phosphate is a suitable probe for studying the active sites of these enzymes. Comparison of the present results with those previously obtained with Eschericha coli RNA polymerase in this laboratory suggest a new degree of structural homology between eucaryotic and procaryotic RNA polymerases.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Martial, Joseph ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Zaldivar, J.
Bull, P.
Venegas, A.
Valenzuela, P.
Language :
English
Title :
Inactivation of rat liver RNA polymerases I and II and yeast RNA polymerase I by pyrodixal 5'-phosphate. Evidence for the participation of lysyl residues at the active site.
Publication date :
04 November 1975
Journal title :
Biochemistry
ISSN :
0006-2960
eISSN :
1520-4995
Publisher :
American Chemical Society, Washington, United States - District of Columbia
scite shows how a scientific paper has been cited by providing the context of the citation, a classification describing whether it supports, mentions, or contrasts the cited claim, and a label indicating in which section the citation was made.
Bibliography
Anderson, B. M., Anderson, C. D., and Churchich, J. E. (1966), Biochemistry 5, 2893.
Buhler, J. M., Sentenac, A., and Fromageot, P. (1974), J. Biol. Chem. 249, 5963.
Bull, P., Zaldivar, J., Venegas, A., Martial, J., and Valenzuela, P. (1975), Biochem. Biophys. Res. Commun. 64, 1152.
Chambon, P. (1970), Enzymes, 3rd Ed. 10, 261-331.
Chesterton, G. J., and Butterworth, P. H. W. (1971), FEBS Lett. 15, 181.
Fischer, E. H., Forrey, A. W., Hedrick, J. L., Hughes, R. C., Kent, A. B., and Krebs, E. G. (1963), in Chemical and Biological Aspects of Pyridoxal Catalysis, Snell, E. E., Fasella, P. M., Braunstein, A., and Rossi-Fanelli, A., Ed., New York, N.Y., Pergamon Press, p 554.
Gissinger, F., and Chambon, P. (1972), Eur. J. Biochem. 28, 277.
Goldberg, M., Perriard, J. C., Hager, G., Hallick, R. E., and Rutter, W. J. (1973), in Control of Transcription, Biswas, B. B., Mandal, R. K., Stevens, A., and Cohn, W. E., Ed., New York, N.Y., Plenum Press, p 241.
Hinkle, D. C., and Chamberlin, M. J. (1972), J. Mol. Biol. 70, 157.
Johnson, G. S., and Deal, W. C. (1970), J. Biol. Chem. 245, 238.
Kedinger, C., and Chambon, P. (1972), Eur. J. Biochem. 28, 283.
Kedinger, C., Gissinger, F., Gniazdowski, M., Mandel, J. L., and Chambon, P. (1972), Eur. J. Biochem. 28, 269.
Kirkegaard, L. H., Johnson, T. J. A., and Bock, R. M. (1972), Anal. Biochem. 50, 122.
Krakow, J. A., and Goolsby, S. P. (1971), Biochem. Biophys. Res. Commun. 44, 453.
Mandel, J. L., and Chambon, P. (1971), FEBS Lett. 15, 175.
Marcus, F., and Hubert, E. (1968), J. Biol. Chem. 243, 4923.
Rippa, M., Spanio, L., and Pontremoli, S. (1967), Arch. Biochem. Biophys. 118, 48.
Roeder, R. G., and Rutter, W. J. (1969), Nature (London) 224, 234.
Roeder, R. G., and Rutter, W. J. (1970), Proc. Natl. Acad. Sci. U.S.A. 65, 675.
Schnackerz, K. D., and Noltman, E. A. (1971), Biochemistry 10, 4837.
Schwartz, L. B., and Roeder, R. G. (1974), J. Biol. Chem. 249, 5898.
Schwartz, L. B., and Roeder, R. G. (1975), J. Biol. Chem. 250, 3221.
Shapiro, S., Enser, M., Pugh, E., and Horecker, B. L. (1968), Arch. Biochem. Biophys. 128, 554.
Valenzuela, P., Bull, P., Martial, J., Venegas, A., Zaldivar, J., and Tellez, R. (1974), Fed. Proc., Fed. Am. Soc. Exp. Biol. 33, 1417.
Valenzuela, P., Morris, R. W., Faras, A., Levinson, W., and Rutter, W. J. (1973), Biochem. Biophys. Res. Commun. 53, 1036.
Venegas, A., Martial, J., and Valenzuela, P. (1973), Biochem. Biophys. Res. Commun. 55, 1053.
Weaver, R. F., Blatti, S. P., and Rutter, W. J. (1971), Proc. Natl. Acad. Sci. U.S.A. 68, 2994.
Widnell, C. C., and Tata, J. R. (1964), Biochem. J. 92, 313.
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