Abstract
Study of protein expression during the cell cycle requires preparation of pure fractions of cells at various phases of the cell cycle. This was achieved by the development of methods for cell synchronization. Successful cell synchronization requires knowledge of the duration of all phases of the cell cycle. So, in the present review these interrelated problems are considered together. The first part of this review deals with basic methods employed for analysis of duration of cell cycle phases. The second summarizes data on treatments used for cell synchronization. Methods for calculation of percent of cells at various stages of the cell cycle in fractions of synchronized cells are considered in the third part. The fourth part of this review deals with a method of study of protein expression during the cell cycle by means of immunoblotting of synchronized cell fractions. In the Appendix, basic principles are illustrated with practical examples of analysis of the cell cycle, synchronization, and study of expression of some proteins at various stages of the cell cycle using synchronized XL2 (Xenopus laevis) cells.
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REFERENCES
Howard, A., and Pelc, S. R. (1953) Heredity Suppl., 6, 261-273.
Zetterberg, A., and Larsson, O. (1995) in Cell Cycle Control (Hutchison, C., and Glover, D., eds.) IRL Press at Oxford University Press, Oxford, pp. 205-227.
Lajtha, L. G. (1963) J. Cell Compar. Physiol., 62, 143-145.
Quastler, H. (1963) Cell Proliferation, Blackwell, Oxford.
Mendelsohn, M. L. (1960) Science, 132, 1496.
Kisieleski, W. E., Baserga, R., and Lisco, H. (1961) Atompraxis, 7, 81-85.
Evans, T., Rosenthal, E. T., Youngblom, J., Distel, D., and Hunt, T. (1983) Cell, 33, 389-396.
Norbury, C., and Nurse, P. (1992) Annu Rev. Biochem., 61, 441-470.
Epifanova, O. I. (1997) Lections about Cell Cycle [in Russian], Commonwealth of Scientific Publishing Houses KMK, Moscow.
Nurse, P. (2000) Cell, 100, 71-78.
Bravo, R., Fey, S. J., Bellatin, J., Larsen, P. M., and Celis, J. E. (1982) Progr. Clin. Biol. Res., 85, Pt. A, 235-248.
Mathews, M. B., Bernstein, R. M., Franza, B. R., Jr., and Garrels, J. I. (1984) Nature, 309, 374-376.
Zimmerman, A. M., Zimmerman, S., Thomas, J., and Ginzburg, I. (1983) FEBS Lett., 164, 318-321.
Weisenberg, R. C. (1972) J. Cell Biol., 54, 266-278.
Hinchcliffe, E. H., Li, C., Thompson, E. A., Maller, J. L., and Sluder, G. (1999) Science, 283, 851-854.
Lacey, K. R., Jackson, P. K., and Stearns, T. (1999) Proc. Natl. Acad. Sci. USA, 96, 2817-2822.
Meraldi, P., Lukas, J., Fry, A. M., Bartek, J., and Nigg, E. A. (1999) Nature Cell Biol., 1, 88-93.
Uzbekov, R., Kireev, I., and Prigent, C. (2002) Biol. Cell, 94, 275-288.
Dawson, K. B., Madoc-Jones, H., and Field, F. G. (1965) Exp. Cell Res., 38, 75-84.
Sakharov, V. N., and Voronkova, L. N. (1993) Tsitologiia, 35, 79-85.
Uzbekov, R. E., Arlot-Bonnemains, Y., Chartrain, I., and Philippe, M. (1998) Biol. Membr. (Moscow), 15, 614-622.
Nias, A. H. W., and Fox, M. (1971) Cell Tissue Kinet., 4, 375-398.
Lodish, H., Baltimore, D., Berk, A., Zipursky, S. L., Matsudaira, P., and Darnell, J. (1995) Molecular Cell Biology, 3rd Edn., W. H. Freeman and Co., N. Y.
Quastler, H., and Sherman, F. G. (1959) Exp. Cell Res., 17, 420-438.
Ley, K. D., and Tobey, R. A. (1970) J. Cell Biol., 47, 453-459.
Zielke, H. R., and Littlefield, J. W. (1974) Meth. Cell Biol., 8, 107-121.
Ikegami, S., Taguchi, T., Ohashi, M., Oguro, M., Nagano, H., and Mano, Y. (1978) Nature (London), 275, 458-460.
Zieve, G. W., Turnbull, D., Mullins, J. M., and McIntosh, J. R. (1980) Exp. Cell Res., 126, 397-405.
Pardee, A. B. (1989) Science, 246, 603-608.
Lalande, M. (1990) Exp. Cell Res., 186, 332-339.
Pardee, A. B., and Keyomarsi, K. (1992) Curr. Opin. Cell Biol., 4, 186-191.
Sherwood, S. W., Kung, A. L., Roitelman, J., Simoni, R. D., and Schimke, R. T. (1993) Proc. Natl. Acad. Sci. USA, 90, 3353-3357.
Yerly-Motta, V., Pavy, J. J., and Herve, P. (1998) Biotech. Histochem., 74, 119-128.
Xeros, N. (1962) Nature (London), 194, 682.
Tobey, R. A., Petersen, D. F., Anderson, E. C., and Puck, T. T. (1966) Biophys. J., 6, 567-581.
Bostock, C. J., Prescott, D. M., and Kirkpatrick, J. B. (1971) Exp. Cell Res., 68, 163-168.
Stein, G. S., and Borun, T. W. (1972) J. Cell Biol., 52, 292-307.
Stein, G. S., Stein, J. L., Lian, J. B., Last, T. J., Owen, T. A., and McCabe, L. (1998) in Cell Biology: a Laboratory Handbook, 2nd Edn., Vol. 1, Academic Press, N. Y., pp. 253-260.
Kasten, F. H., Strasser, F. F., and Turner, M. (1965) Nature (London), 207, 161-164.
Pedrali-Noy, G., and Spadari, S. (1979) Biochem. Biophys. Res. Commun., 88, 1194-1202.
Pedrali-Noy, G., Spadari, S., Miller-Faures, A., Miller, A. O., Kruppa, J., and Koch, G. (1980) Nucleic Acids Res., 8, 377-387.
Matherly, L. H., Schuetz, J. D., Westin, E., and Goldman, I. D. (1989) Analyt. Biochem., 182, 338-345.
Fox, M. H., Read, R. A., and Bedford, J. S. (1987) Cytometry, 8, 315-320.
Uzbekov, R., Chartrain, I., Philippe, M., and Arlot-Bonnemains, Y. (1998) Exp. Cell Res., 242, 60-68.
Uzbekov, R., Prigent, C., and Arlot-Bonnemains, Y. (1999) Micr. Res. Tech., 45, 31-42.
Lonn, U., and Lonn, S. (1983) Proc. Natl. Acad. Sci. USA, 80, 3996-3999.
Iliakis, G., Nusse, M., and Bryant, P. (1982) Int. J. Radiat. Stud. Phys. Chem. Med., 42, 417-434.
Cordeiro-Stone, M., and Kaufman, D. G. (1985) Biochemistry, 24, 4815-4822.
Radford, I. R., and Broadhurst, S. (1988) Int. J. Radiat. Stud. Phys. Chem. Med., 53, 205-215.
D'Anna, J. A., Crissman, H. A., Jackson, P. J., and Tobey, R. (1985) Biochemistry, 24, 5020-5026.
Reichard, P., and Ehrenberg, A. (1983) Science, 221, 514-519.
Nakamura, M., Saito, H., Ebinuma, H., Wakabayashi, K., Saito, Y., Takagi, T., Nakamoto, N., and Ishii, H. (2001) J. Cell Physiol., 187, 392-401.
Johnson, R. T., Downes, C. S., and Meyn, R. E. (1993) in The Cell Cycle. A Practical Approach (Fantes, P., and Brooks, R., eds.) Oxford University Press, N. Y., pp. 1-24.
Urbani, L., Sherwood, S., and Schimke, R. T. (1995) Exp. Cell. Res., 219, 159-168.
Keyomarsi, K., Sandoval, L., Band, V., and Pardee, A. B. (1991) Cancer Res., 51, 3602-3609.
Wu Jia-Rui, and Gilbert, D. M. (2000) FEBS Lett., 484, 108-112.
Jakobisiak, M., Bruno, S., Skierski, J. S., and Darzynkiewicz, A. (1991) Proc. Natl. Acad. Sci. USA, 88, 3628-3632.
Stryer, L. (1981) Biochemistry, 2nd Edn., W. H. Freeman and Co., San Francisco.
Yunis, J. J. (1976) Science, 191, 1268-1270.
Pai, G. S., and Tomas, G. H. (1980) Hum. Genet., 54, 41-45.
Mosca, P. J., Dijkwel, P. A., and Hamlin, R. N. (1992) Mol. Cell Biol., 12, 4375-4383.
Gilbert, D. M., Neilson, A., Miyazawa, H., DePamphilis, M. L., and Burhans, W. C. (1995) J. Biol. Chem., 270, 9597-9606.
Kalejta, R. F., and Hamlin, J. L. (1997) Exp. Cell Res., 231, 173-183.
Hoffman, B. D., Hanauske-Abel, H. M., Flint, A., and Lalande, M. (1991) Cytometry, 12, 26-32.
De Brabander, M. J., van de Veire, R. M. L., Aerts, F. E. M., Borgers, M., and Janssen, P. A. J. (1976) Cancer Res., 36, 905-916.
Quintart, J., Bartholeyns, J., and Baudhuin, P. (1979) Biochem. J., 184, 133-141.
Rieder, C. L., and Cole, R. (2000) Curr. Biol., 10, 1067-1070.
Nusse, M., and Egner, H. J. (1984) Cell Tissue Kinet., 17, 13-23.
Kwon, T. K., Buchholz, A., Chrest, F. J., and Nordin, A. A. (1996) Cell Growth Differ., 7, 1305-1313.
Schnier, J. B., Nishi, K., Goodrich, D. W., and Bradbury, E. M. (1996) Proc. Natl. Acad. Sci. USA, 93, 5941-5946.
Courage, C., Snowden, R., and Gescher, A. (1996) Br. J. Cancer, 74, 1199-1205.
Shimizu, E., Zhao, M. R., Nakanishi, H., Yamamoto, A., Yoshida, S., Takada, M., Ogura, T., and Sone, S. (1996) Oncology, 53, 494-504.
Swe, M., Bay, B. H., and Sit, K. H. (1996) Cancer Lett., 104, 145-152.
Temin, H. (1971) J. Cell Physiol., 78, 161-170.
Pardee, A. B. (1974) Proc. Natl. Acad. Sci. USA, 71, 1286-1290.
Prescott, D. M. (1976) Reproduction of Eukaryote Cells, Academic Press, N. Y.
Niki, T., Rombouts, K., De Bleser, P., de Smet, K., Rogiers, V., Schuppan, D., Yoshida, M., Gabbiani, G., and Geerts, A. (1999) Hepatology, 29, 858-867.
Yoshida, M., and Beppu, T. (1988) Exp. Cell Res., 177, 122-131.
Jackson, R. S. (1978) J. Biol. Chem., 253, 7440-7446.
De Braekeleer, M., Keushnig, M., and Lin, C. C. (1985) Can. J. Genet. Cytol., 27, 622-625.
Lalande, M., and Hanauske-Abel, H. M. (1990) Exp. Cell Res., 188, 117-121.
Jue, S. G., Dawson, G. W., and Brogden, R. N. (1985) Drugs, 29, 330-341.
Tournier, F., Karsenti, E., and Bornens, M. (1989) Dev. Biol., 136, 321-329.
Chen, G., and Hitomi, M. (1999) Exp. Cell Res., 249, 327-336.
Shall, S. (1973) Meth. Cell Biol., 7, 269-285.
Mitchell, B. F., and Tupper, J. T. (1977) Exp. Cell Res., 106, 351-355.
Kaiser, N., Bourne, H. R., Insel, P. A., and Coffino, P. (1979) J. Cell Phys., 101, 369-374.
Gilbert, D. M., and Cohen, S. N. (1987) Cell, 50, 59-68.
Monahan, J. J. (1976) in Methods in Cell Biology, Vol. 14 (Prescott, D. M., ed.) Academic Press, N. Y., pp. 105-111.
Rice, G. C., Dean, P. N., Gray, J. W., and Dewey, W. C. (1984) Cytometry, 5, 289-298.
Crissman, H. A., and Tobey, R. A. (1974) Science, 184, 1297-1298.
Vindelov, L. L., Christensen, I. J., and Nissen, N. I. (1983) Cytometry, 3, 328-331.
Burhans, W. C., Vassilev, L. T., Caddle, M. S., Heintz, N. H., and DePamphilis, M. L. (1990) Cell, 62, 955-965.
Dolbeare, F., Gratzner, H., Pallavicini, M., and Gray, J. W. (1983) Proc. Natl. Acad. Sci. USA, 80, 5573-5577.
Gorbsky, G. J. (1994) Cancer Res., 54, 1042-1048.
Kamentsky, L. A., and Kamentsky, L. D. (1991) Cytometry, 12, 381-387.
Kamentsky, L. A., Burger, D. E., Gershman, R. J., Kamentsky, L. D., and Luther, E. (1997) Acta Cytol., 41, 123-143.
Epifanova, O. I., Terskikh, V. V., and Zakharov, A. F. (1977) Radioautography [in Russian], Vysshaya Shkola, Moscow.
Gratzner, H. G. (1982) Science, 218, 474-475.
Vorobiev, I. A., and Nadezhdina, E. S. (1987) Advances in Science and Technology. General Problems of Physico-Chemical Biology, Vol. 7 (Chentsov, Yu. S., ed.) [in Russian], VINITI, Moscow, pp. 1-164.
Uzbekov, R. E. (2004) Tsitologiya, 45, 249-256.
Uzbekov, R. E., Timirbulatova, E. R., Watrin, E., Cubizolles, F., Ogereau, D., Gulak, P., Legagneux, V., Polyakov, V. Ju., Le Guellec, K., and Kireev, I. (2003) J. Cell Sci., 116, 1667-1678.
Roghi, C., Giet, R., Uzbekov, R., Morin, N., Chartrain, I., Le Guellec, R., Couturier, A., Doree, M., Philippe, M., and Prigent, C. (1998) J. Cell Sci., 111, 157-172.
Arlot-Bonnemains, Y., Klotzbucher, A., Giet, R., Uzbekov, R., Bihan, R., and Prigent, C. (2001) FEBS Lett., 508, 149-152.
Uzbekov, R. E., Giet, R., Arlot-Bonnemains, Y., and Prigent, C. (2001) Biol. Cell, 93, 337.
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Uzbekov, R.E. Analysis of the Cell Cycle and a Method Employing Synchronized Cells for Study of Protein Expression at Various Stages of the Cell Cycle. Biochemistry (Moscow) 69, 485–496 (2004). https://doi.org/10.1023/B:BIRY.0000029845.11184.30
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DOI: https://doi.org/10.1023/B:BIRY.0000029845.11184.30