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Regulation of human thymidine kinase during the cell cycle.

Open AccessPublished:June 15, 1988DOI:https://doi.org/10.1016/S0021-9258(18)68484-4
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      As an approach to defining the molecular basis for the periodic expression of thymidine kinase activity during the cell cycle, we have examined properties of the cytosolic enzyme in cycling HeLa cells synchronized by centrifugal elutriation and mitotic selection. By immunoblot analyses with a specific antiserum raised against the purified HeLa enzyme, we have demonstrated that changes in the levels of thymidine kinase activity reflect similar changes in the levels of thymidine kinase polypeptide. In contrast, the steady state levels of thymidine kinase mRNA show relatively small changes during the cell cycle. Using pulse labeling methods, we have shown that the synthetic rate of thymidine kinase protein is about 10-fold greater in S phase than in G1 phase, indicating that the efficiency of translation of thymidine kinase mRNA increases as cells begin DNA replication. In addition, the stability of thymidine kinase protein dramatically decreases upon cell division, resulting in the rapid clearance of the enzyme from newly divided G1 cells. Thus, two different post-transcriptional mechanisms largely account for the periodic behavior of the enzyme activity during the cell cycle.

      REFERENCES

        • Bello L.J.
        Cell Res. 1974; 89: 263-274
        • Bradford M.M.
        Anal. Biochem. 1976; 72: 248-254
        • Bradshaw Jr., H.D.
        Proc. Natl. Acad. Sci. U. S. A. 1983; 80: 5588-5591
        • Bradshaw Jr., H.D.
        • Deininger P.L.
        Mol. Cell. Biol. 1984; 4: 2316-2320
        • Brent T.P.
        Cell Tissue Kinet. 1971; 4: 297-305
        • Brown E.H.
        • Schildkraut C.L.
        J. Cell. Physiol. 1979; 99: 261-278
        • Coppock D.L.
        • Pardee A.B.
        Mol. Cell. Biol. 1987; 7: 2925-2932
        • Evans T.
        • Rosenthal E.T.
        • Youngblom J.
        • Distel D.
        • Hunt T.
        Cell. 1983; 33: 389-396
        • Farnham P.J.
        • Schimke R.T.
        J. Biol. Chem. 1985; 260: 7675-7680
        • Farnham P.J.
        • Schimke R.T.
        Mol. Cell. Biol. 1986; 6: 365-371
        • Feinberg A.P.
        • Vogelstein B.
        Anal. Biochem. 1983; 132: 6-13
        • Hendrickson S.L.
        • Wu J.-S. R.
        • Johnson L.F.
        Proc. Natl. Acad. Sci. U. S. A. 1980; 77: 5140-5144
        • Hochauser S.J.
        • Stein J.L.
        • Stein G.S.
        Int. Rev. Cytol. 1981; 71: 95-243
        • Jenh C.-H.
        • Geyer P.K.
        • Johnson L.F.
        Mol. Cell. Biol. 1985; 5: 2527-2532
        • Jenh C.-H.
        • Rao L.G.
        • Johnson L.F.
        J. Cell. Physiol. 1985; 122: 149-154
        • Johnson L.F.
        • Rao L.G.
        • Muench A.J.
        Exp. Cell Res. 1982; 138: 79-85
        • Kit S.
        Mol. Cell. Biochem. 1976; 11: 161-182
        • Kit S.
        • Leung W.-C.
        • Trkula D.
        Arch. Biochem. Biophys. 1973; 158: 503-513
        • Kreidberg J.A.
        • Kelly T.J.
        Mol. Cell. Biol. 1986; 6: 2903-2909
        • Kucera R.
        • Brown C.L.
        • Paulus H.
        J. Cell. Physiol. 1983; 117: 158-168
        • Lachman H.M.
        • Hatton K.S.
        • Skoultchi A.I.
        • Schildkraut C.L.
        Proc. Natl. Acad. Sci. U. S. A. 1985; 82: 5323-5327
        • Lewis J.A.
        • Shimizu K.
        • Zipser D.
        Mol. Cell. Biol. 1983; 3: 1815-1823
        • Maniatis T.
        • Fritsch E.F.
        • Sambrook J.
        Molecular Cloning: A Laboratory Manual.
        Cold Spring Harbor Laboratory, Cold Spring Harbor, NY1982: 202
        • Mariani B.D.
        • Slate D.L.
        • Schimke R.T.
        Proc. Natl. Acad. Sci. U. S. A. 1981; 78: 4985-4989
        • Paul J.R.
        Cell and Tissue Culture.
        Williams and Wilkins Co., Baltimore, MD1960: 253
        • Ricciardi R.P.
        • Miller J.S.
        • Roberts B.E.
        Proc. Natl. Acad. Sci. U. S. A. 1979; 76: 4927-4931
        • Robbins E.
        • Marcus P.I.
        Science. 1964; 144: 1152-1153
        • Schlosser C.A.
        • Steglich C.
        • DeWet J.R.
        • Scheffler I.E.
        Proc. Natl. Acad. Sci. U. S. A. 1981; 78: 1119-1123
        • Sherley J.L.
        Towards Understanding the Control of Chromosomal DNA Replication: Studies on the Human Cytosolic Thymidine Kinase.
        (Ph.D. Dissertation) Johns Hopkins University, 1988
        • Sherley J.L.
        • Kelly T.J.
        J. Biol. Chem. 1988; 263: 375-382
        • Stewart C.J.
        • Ito M.
        • Conrad S.E.
        Mol. Cell. Biol. 1987; 7: 1156-1163
        • Stuart P.
        • Ito M.
        • Stewart C.
        • Conrad S.E.
        Mol. Cell. Biol. 1985; 5: 1490-1497
        • Swenson K.I.
        • Farrell K.M.
        • Ruderman J.V.
        Cell. 1986; 47: 861-870
        • Thömmes P.
        • Reiter T.
        • Knippers R.
        Biochemistry. 1986; 25: 1308-1314
        • Tricoli J.V.
        • Sahai B.M.
        • McCormick P.J.
        • Jarlinski S.J.
        • Bertram J.S.
        • Kowalski D.
        Exp. Cell Res. 1985; 158: 1-14
        • Wiedemann L.M.
        • Johnson L.F.
        Proc. Natl. Acad. Sci. U. S. A. 1979; 76: 2818-2822