Abstract
Heterogeneous nuclear ribonucleoprotein particles (HnRNP) were separated in metrizamide density gradients, into two fractions migrating to 1.31 g ml-1 and 1.18 g ml-1, respectively. Proteins associated with each of these fractions were analysed by SDS-acrylamide gel electrophoresis. It is shown that the whole proteins extracted from these two metrizamide fractions exhibit clearly different electrophoretic patterns: 1.31 HnRNP particles contain as major polypeptide chains molecules with molecular weights ranging from 40 000 to 65 000, while major polypeptides of 1.18 HnRNP are banding in the 30 000–40 000 molecular weight region of the gels. Both fractions contain numerous other associated polypeptide chains whose molecular weights are above 65 000. A possible kinetic relationship between these two HnRNP classes was investigatedin vivo by performing chase experiments. No clear evidence for a precursor-product relationship was found. Implications arising from these structural and kinetic observations, and problems relating to nuclear maturation of pre-messenger RNA, are discussed.
Similar content being viewed by others
References
Harris, H.,Biochem. J. 84, 60 (1962).
Houssais, J. F. and Attardi, G.,Proc. Nat. Acad. Sci. U.S.A. 56, 616 (1966).
Soeiro, R., Birboim, H. and Darnell, J. E.,J. Mol. Biol. 19, 362 (1966).
Scherrer, K., Marcaud, L., Zajdela, F., London, I. M., and Gros, F.,Proc. Nat. Acad. Sci. U.S.A. 56, 1571 (1966).
Spohr, G., Imaizumi, T., and Scherrer, K.,Proc. Nat. Acad. Sci. U.S.A. 71, 5009 (1974).
Imaizumi, T., Diggelman, H., and Scherrer, K.,Proc. Nat. Acad. Sci. U.S.A. 70, 1122 (1973).
Williamson, R., Drewienkiewicz, C. E., and Paul, J.,Nature New Biol. 241, 66 (1973).
Ruiz-Carillo, A., Beato, M., Schutz, G., Feigelson, P., and Allfrey, V. G.,Proc. Nat. Acad. Sci. U.S.A. 70, 3641 (1973).
Macnaughton, M., Freeman, K. B., and Bishop, J. O.,Cell 1, 117 (1974).
Price, R. P., Ransom, L., and Penman, S.,Cell 2, 253 (1974).
Spohr, G., Dettori, G., and Manzari, V.,Cell 8, 505 (1976).
Hendrick, D. and De Kloet, S. R.,Mol. Biol. Reports 2, 225 (1975).
Samarina, O. P., Lukanidin, E. M., Molnar, J., and Georgiev, G. P.,J. Mol. Biol. 33, 251 (1968).
Stevenin, J., Mandel, P., and Jacob, M.,Bull. Soc. Chim. Biol. 52, 703 (1970).
Niessing, J. and Sekeris, C. E.,Biochim. Biophys. Acta 247, 391 (1971).
Faiferman, I., Hamilton, M. G., and Pogo, A. O.,Biochim. Biophys. Acta 232, 685 (1971).
Sommerville, J. and Hill, R. J.,Nature New Biol. 245, 104 (1973).
Albrecht, C. and Van Zyl, I. M.,Exp. Cell Res. 76, 8 (1973).
Pederson, T.,J. Mol. Biol. 83, 163 (1974).
Ducamp, C. and Jeanteur, P.,Biochimie 55, 1235 (1973).
Faiferman, I. and Pogo, A. O.,Biochemistry 14, 3808 (1975).
Kumar, A. and Pederson, T.,J. Mol. Biol. 96, 353 (1975).
Houssais, J. F.,FEBS Letters 56, 341 (1975).
Houssais, J. F., inBiological Separation in Iodinated Density Gradient Media, D. Rickwood (ed.), Information Retrieval Limited, London and Washington, D.C., (1976) p. 81.
Matringe, H. and Jacob, M.,Biochimie 54, 1169 (1972).
Krichevskaya, A. A. and Georgiev, G. P.,Biochim. Biophys. Acta 194, 619 (1969).
Martin, T., Billings, P., Levey, A., Ozarslan, S., Quinlan, T., Swift, H., and Urbas, L.,Cold Spring Harbor Symp. Quant. Biol. 38, 921 (1974).
Osborne-Rivet, L. and Houssais, J.F.,Eur. J. Biochem. 48, 427 (1974).
Houssais, J. F.Eur. J. Biochem. 24, 232 (1971).
Laemmli, U.K.,Nature 227, 680 (1970).
Gallinaro-Matringe, H., Stevenin, J., and Jacob, M.,Biochemistry 14, 2547 (1975).
Stevenin, J., Gallinaro-Matringe, H., Gattoni, R., and Jacob, M.,J. Mol. Biol., in press.
Ross, J.,J. Mol. Biol. 106, 403 (1976).
Kinniburgh, A. J. and Martin, T. E.,Proc. Nat. Acad. Sci. U.S.A. 73, 2725 (1976).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Houssais, JF. A comparative study on the two classes of heterogeneous nuclear ribonucleoprotein particles separated in metrizamide density gradient, by electrophoresis of proteins and chase experiments. Evidence for two distinct subfractions of HnRNP in mammalian nuclei. Mol Biol Rep 3, 251–261 (1977). https://doi.org/10.1007/BF00643481
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00643481