Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Differences in mutant p53 protein stability and functional activity in teniposide-sensitive and -resistant human leukemic CEM cells

Abstract

We examined p53 protein stability and DNA damage-induced p53-dependent responses in a human leukemic CEM cell line and two teniposide-resistant sublines, CEM/VM-1 and CEM/VM-1-5 (40 and 400-fold resistant to teniposide, respectively). Although all cell lines contain the same p53 mutations at codons 175 (Arg→His) and 248 (Arg→Gln), the constitutive levels of p53 were progressively increased with the resistance of the cells to teniposide. By pulse-chase experiments, we found that the half-lives of mutant p53 protein were approximately 12, 17, and >30 h in CEM, CEM/VM-1, and CEM/VM-1-5 cells, respectively. The prolonged half-lives of p53 in these cells is consistent with the fact that the protein harbors the indicated mutations. Of note, however, is the fact that the increased p53 protein half-lives in the two drug-resistant cell lines corresponds to a proportional decrease in MDM2 protein levels but an increase in p53-MDM2 binding interactions. This suggests that MDM2-mediated p53 degradation may be altered in our leukemic cell lines. The DNA damage-induced p53 response is fully functional in the drug-sensitive CEM cells containing a mutant p53, but this pathway is attenuated in the drug-resistant cells. Specifically, while the mutant p53 was phosphorylated at serine-15 in response to ionizing radiation in all these cell lines, mutant p53 induction in response to teniposide or ionizing radiation and induction of the p53-target genes, p21 and GADD45 only occurred in the drug-sensitive CEM cells. As assessed by MTT cytotoxicity assay, CEM cells were also significantly more sensitive to ionizing radiation, compared to the drug-resistant cell lines, and this correlated with p53 induction. Collectively, these results suggest that changes in constitutive mutant p53 protein levels, p53-MDM2 binding interactions, and altered regulation of the DNA damage-inducible p53-dependent pathway may play a role in drug- and radiation-responsiveness in these cells.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9

Similar content being viewed by others

References

  • Alarcon R, Koumenis C, Geyer RK, Maki C and Giaccia AJ. . 1999 Cancer Res. 59: 6046–6051.

  • Arriola EL, Lopez AR and Chresta CM. . 1999 Oncogene 18: 1081–1091.

  • Barak Y, Juven T, Haffner R and Oren M. . 1993 EMBO J. 12: 461–468.

  • Boise LH, Gottschalk AR, Quintans J and Thompson CB. . 1995 Curr. Topics. Microbiol. Immunol. 200: 107–121.

  • Brown R, Clugston C, Burns P, Edlin A, Vasey P, Vojtesek B and Kaye SB. . 1993 Int. J. Cancer 55: 678–684.

  • Bugg BY, Danks MK, Beck WT and Suttle DP. . 1991 Proc. Natl. Acad. Sci. USA 88: 7654–7658.

  • Chen M and Beck WT. . 1993 Cancer Res. 53: 5946–5953.

  • Cheng EHJ, Kirsch DG, Clem RJ, Ravi R, Kastan MB, Bedi A, Ueno K and Hardwick JM. . 1997 Science 278: 1966–1968.

  • Cheng J and Haas M. . 1990 Mol. Cell. Biol. 10: 5502–5509.

  • Craig RW. . 1995 Semin. Cancer Biol. 6: 35–43.

  • Danks MK, Schmidt CA, Cirtain MC, Suttle DP and Beck WT. . 1988 Biochemistry 27: 8861–8869.

  • Danks MK, Warmoth MR, Friche E, Granzen B, Bugg BY, Harker WG, Zwelling LA, Futshcer BW, Suttle DP and Beck WT. . 1993 Cancer Res. 53: 1373–1379.

  • Danks MK, Yalowich J and Beck WT. . 1987 Cancer Res. 47: 1297–1301.

  • Donehower LA and Bradley A. . 1993 Biochem. Biophys. Acta 1155: 181–205.

  • El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW and Vogelstein B. . 1993 Cell 75: 817–825.

  • Eliyahu D, Michalovitz D, Eliyahu S, Pinhasi-Kimhi O and Oren M. . 1989 Proc. Natl. Acad. Sci. USA 86: 8763–8767.

  • Finlay CA, Hinds PW and Levine AJ. . 1989 Cell 57: 1083–1093.

  • Foster BA, Coffey HA, Morin MJ and Rastinejad F. . 1999 Science 286: 2507–2510.

  • Giaccia AJ and Kastan MB. . 1998 Genes. Dev. 12: 2973–2983.

  • Harper JW, Adami GR, Wei N, Keyomarsi K and Elledge SJ. . 1993 Cell 75: 805–816.

  • Hartwell LH and Kastan MB. . 1994 Science 266: 1821–1828.

  • Haupt Y, Maya R, Kazaz A and Oren M. . 1997 Nature 387: 296–299.

  • Hollstein M, Sidransky D, Vogelstein B and Harris CC. . 1991 Science 253: 49–53.

  • Honda R, Tanaka H and Yasuda H. . 1997 FEBS Lett. 420: 25–27.

  • Kastan MB, Onyekwere O, Sidransky D, Vogelstein B and Craig RW. . 1991 Cancer Res. 51: 6304–6311.

  • Kastan MB, Zhan Q, El-Diery WS, Carrier F, Jacks T, Walsh WV, Plunkett BS, Vogelstein B and Fornace AJ. . 1992 Cell 71: 587–597.

  • Kim R and Beck WT. . 1994 Cancer Res. 54: 4958–4966.

  • Kubbutat MHG, Jones SN and Vousden KH. . 1997 Nature 387: 299–303.

  • Kubbutat MHG, Ludwig RL, Levine AJ and Vousden KH. . 1999 Cell Growth Differ. 10: 87–92.

  • Kwok TT, Mok CH and Menton-Brennan L. . 1994 Cancer Res. 54: 2834–2836.

  • Laemmli UK. . 1970 Nature (Lond.) 227: 680–685.

  • Maki CG, Huibregtse JM and Howley PM. . 1996 Cancer Res. 56: 2649–2654.

  • Maltzman W and Czyzyk L. . 1984 Mol. Cell. Biol. 4: 1689–1694.

  • Miyashita T, Krajewski S, Krajewska M, Wang HG, Lin HK, Liebermann DA, Hoffman B and Reed JC. . 1994 Oncogene 9: 1799–1805.

  • Miyashita T and Reed JC. . 1995 Cell 80: 293–299.

  • Momand J, Zambetti GP, Olson DC, George DL and Levine AJ. . 1992 Cell 69: 1237–1245.

  • Morgan SE and Kastan MB. . 1997 Adv. Cancer Res. 71: 1–25.

  • Oltvai ZN, Milliman CL and Korsmeyer SJ. . 1993 Cell 74: 609–619.

  • Park DJ, Nakamura H, Chumakov AM, Said JW, Miller CW, Chen DL and Koeffler HP. . 1994 Oncogene 9: 1899–1906.

  • Prives C. . 1998 Cell 95: 5–8.

  • Reed JC. . 1994 J. Cell. Biol. 124: 1–6.

  • Sedlak TW, Oltvai ZN, Yang E, Wang K, Boise LH, Thompson CB and Korsmeyer SJ. . 1995 Proc. Natl. Acad. Sci. USA 92: 7834–7838.

  • Selvakumaran M, Lin H, Miyashita T, Wang HG, Krajewski S, Reed JC, Hoffman B and Liebermann D. . 1994 Oncogene 9: 1791–1798.

  • Siliciano JD, Canman CE, Taya Y, Sakaguchi K, Appella E and Kastan MB. . 1997 Genes. Dev. 11: 3471–3481.

  • Smith ML, Chen I-T, Zhan Q, Bae I, Chen C-Y, Gilmer TM, Kastan MB, O'Connor PM and Fornace Jr AJ. . 1994 Science 266: 1376–1380.

  • Tishler RB, Calderwood SK, Coleman CN and Price BD. . 1993 Cancer Res. 53: 2212–2216.

  • Ullrich SJ, Anderson CW, Mercer WE and Appella E. . 1992 J. Biol. Chem. 267: 15259–15262.

  • Wang Q and Beck WT. . 1998 Cancer Res. 58: 5762–5769.

  • Zambetti GP and Levine AJ. . 1993 FASEB J. 7: 855–865.

  • Zauberman A, Barak Y, Ragimov N, Levy N and Oren M. . 1993 EMBO J. 12: 2799–2808.

  • Zhang W, Funk WD, Wright WE, Shay JW and Deisseroth AB. . 1993 Oncogene 8: 2555–2559.

Download references

Acknowledgements

We are grateful to Linda Rawlinson and the Biomedical Communications Department at St. Jude Children's Research Hospital for preparation of some of the artwork. This work was supported in part by Research Grants CA40570 and CA30103 (to WT Beck) from the National Cancer Institute, DHHS, Bethesda, MD, and in part by the University of Illinois at Chicago.

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Morgan, S., Kim, R., Wang, P. et al. Differences in mutant p53 protein stability and functional activity in teniposide-sensitive and -resistant human leukemic CEM cells. Oncogene 19, 5010–5019 (2000). https://doi.org/10.1038/sj.onc.1203865

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1203865

Keywords

This article is cited by

Search

Quick links