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. 1996 Aug 15;15(16):4392–4401.

Translational regulation of human p53 gene expression.

L Fu 1, M D Minden 1, S Benchimol 1
PMCID: PMC452163  PMID: 8861966

Abstract

In blast cells obtained from patients with acute myelogenous leukemia, p53 mRNA was present in all the samples examined while the expression of p53 protein was variable from patient to patient. Mutations in the p53 gene are infrequent in this disease and, hence, variable protein expression in the majority of the samples cannot be accounted for by mutation. In this study, we examined the regulation of p53 gene expression in human leukemic blasts and characterized the p53 transcripts in these cells. We found control both at the level of RNA abundance and at the level of translation. Four experiments point towards translational control of human p53 gene expression. First, there is no correlation between the level of p53 mRNA and the level of p53 protein expression in blast cells. Second, in two cell lines with similar levels of p53 protein expression but with different levels of p53 mRNA, we find that there is preferential association of p53 mRNA with large polysomes in the cells with less p53 RNA. Third, translation of synthetic human p53 transcripts in cell-free extracts is inhibited by the p53 3'UTR. Fourth, the p53 3'UTR, when present in cis, can repress translation of a heterologous transcript. These observations raise the possibility that human p53 mRNA translation may be regulated in vivo by RNA binding factors acting on the p53 3'UTR.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Banks L., Matlashewski G., Crawford L. Isolation of human-p53-specific monoclonal antibodies and their use in the studies of human p53 expression. Eur J Biochem. 1986 Sep 15;159(3):529–534. doi: 10.1111/j.1432-1033.1986.tb09919.x. [DOI] [PubMed] [Google Scholar]
  2. Bienz-Tadmor B., Zakut-Houri R., Libresco S., Givol D., Oren M. The 5' region of the p53 gene: evolutionary conservation and evidence for a negative regulatory element. EMBO J. 1985 Dec 1;4(12):3209–3213. doi: 10.1002/j.1460-2075.1985.tb04067.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bienz B., Zakut-Houri R., Givol D., Oren M. Analysis of the gene coding for the murine cellular tumour antigen p53. EMBO J. 1984 Sep;3(9):2179–2183. doi: 10.1002/j.1460-2075.1984.tb02110.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Evans T. C., Crittenden S. L., Kodoyianni V., Kimble J. Translational control of maternal glp-1 mRNA establishes an asymmetry in the C. elegans embryo. Cell. 1994 Apr 22;77(2):183–194. doi: 10.1016/0092-8674(94)90311-5. [DOI] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  7. Fenaux P., Jonveaux P., Quiquandon I., Laï J. L., Pignon J. M., Loucheux-Lefebvre M. H., Bauters F., Berger R., Kerckaert J. P. P53 gene mutations in acute myeloid leukemia with 17p monosomy. Blood. 1991 Oct 1;78(7):1652–1657. [PubMed] [Google Scholar]
  8. Fenaux P., Preudhomme C., Quiquandon I., Jonveaux P., Laï J. L., Vanrumbeke M., Loucheux-Lefebvre M. H., Bauters F., Berger R., Kerckaert J. P. Mutations of the P53 gene in acute myeloid leukaemia. Br J Haematol. 1992 Feb;80(2):178–183. doi: 10.1111/j.1365-2141.1992.tb08897.x. [DOI] [PubMed] [Google Scholar]
  9. Feng S., Holland E. C. HIV-1 tat trans-activation requires the loop sequence within tar. Nature. 1988 Jul 14;334(6178):165–167. doi: 10.1038/334165a0. [DOI] [PubMed] [Google Scholar]
  10. Fort P., Marty L., Piechaczyk M., el Sabrouty S., Dani C., Jeanteur P., Blanchard J. M. Various rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family. Nucleic Acids Res. 1985 Mar 11;13(5):1431–1442. doi: 10.1093/nar/13.5.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fritsche M., Haessler C., Brandner G. Induction of nuclear accumulation of the tumor-suppressor protein p53 by DNA-damaging agents. Oncogene. 1993 Feb;8(2):307–318. [PubMed] [Google Scholar]
  12. Fu L. N., Ye R. Q., Browder L. W., Johnston R. N. Translational potentiation of messenger RNA with secondary structure in Xenopus. Science. 1991 Feb 15;251(4995):807–810. doi: 10.1126/science.1990443. [DOI] [PubMed] [Google Scholar]
  13. Gavis E. R., Lehmann R. Translational regulation of nanos by RNA localization. Nature. 1994 May 26;369(6478):315–318. doi: 10.1038/369315a0. [DOI] [PubMed] [Google Scholar]
  14. Goodwin E. B., Okkema P. G., Evans T. C., Kimble J. Translational regulation of tra-2 by its 3' untranslated region controls sexual identity in C. elegans. Cell. 1993 Oct 22;75(2):329–339. doi: 10.1016/0092-8674(93)80074-o. [DOI] [PubMed] [Google Scholar]
  15. Harlow E., Crawford L. V., Pim D. C., Williamson N. M. Monoclonal antibodies specific for simian virus 40 tumor antigens. J Virol. 1981 Sep;39(3):861–869. doi: 10.1128/jvi.39.3.861-869.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harlow E., Williamson N. M., Ralston R., Helfman D. M., Adams T. E. Molecular cloning and in vitro expression of a cDNA clone for human cellular tumor antigen p53. Mol Cell Biol. 1985 Jul;5(7):1601–1610. doi: 10.1128/mcb.5.7.1601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Herzog H., Höfferer L., Schneider R., Schweiger M. cDNA encoding the human homologue of rat ribosomal protein L35a. Nucleic Acids Res. 1990 Aug 11;18(15):4600–4600. doi: 10.1093/nar/18.15.4600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hsu H. C., Tseng H. J., Lai P. L., Lee P. H., Peng S. Y. Expression of p53 gene in 184 unifocal hepatocellular carcinomas: association with tumor growth and invasiveness. Cancer Res. 1993 Oct 1;53(19):4691–4694. [PubMed] [Google Scholar]
  19. Huarte J., Stutz A., O'Connell M. L., Gubler P., Belin D., Darrow A. L., Strickland S., Vassalli J. D. Transient translational silencing by reversible mRNA deadenylation. Cell. 1992 Jun 12;69(6):1021–1030. doi: 10.1016/0092-8674(92)90620-r. [DOI] [PubMed] [Google Scholar]
  20. Jackson R. J. Cytoplasmic regulation of mRNA function: the importance of the 3' untranslated region. Cell. 1993 Jul 16;74(1):9–14. doi: 10.1016/0092-8674(93)90290-7. [DOI] [PubMed] [Google Scholar]
  21. Kastan M. B., Onyekwere O., Sidransky D., Vogelstein B., Craig R. W. Participation of p53 protein in the cellular response to DNA damage. Cancer Res. 1991 Dec 1;51(23 Pt 1):6304–6311. [PubMed] [Google Scholar]
  22. Kastan M. B., Radin A. I., Kuerbitz S. J., Onyekwere O., Wolkow C. A., Civin C. I., Stone K. D., Woo T., Ravindranath Y., Craig R. W. Levels of p53 protein increase with maturation in human hematopoietic cells. Cancer Res. 1991 Aug 15;51(16):4279–4286. [PubMed] [Google Scholar]
  23. Kupiec J. J., Giron M. L., Vilette D., Jeltsch J. M., Emanoil-Ravier R. Isolation of high-molecular-weight DNA from eukaryotic cells by formamide treatment and dialysis. Anal Biochem. 1987 Jul;164(1):53–59. doi: 10.1016/0003-2697(87)90366-6. [DOI] [PubMed] [Google Scholar]
  24. Kwon Y. K., Hecht N. B. Binding of a phosphoprotein to the 3' untranslated region of the mouse protamine 2 mRNA temporally represses its translation. Mol Cell Biol. 1993 Oct;13(10):6547–6557. doi: 10.1128/mcb.13.10.6547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lai J. L., Preudhomme C., Zandecki M., Flactif M., Vanrumbeke M., Lepelley P., Wattel E., Fenaux P. Myelodysplastic syndromes and acute myeloid leukemia with 17p deletion. An entity characterized by specific dysgranulopoïesis and a high incidence of P53 mutations. Leukemia. 1995 Mar;9(3):370–381. [PubMed] [Google Scholar]
  26. Lamb P., Crawford L. Characterization of the human p53 gene. Mol Cell Biol. 1986 May;6(5):1379–1385. doi: 10.1128/mcb.6.5.1379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lu X., Lane D. P. Differential induction of transcriptionally active p53 following UV or ionizing radiation: defects in chromosome instability syndromes? Cell. 1993 Nov 19;75(4):765–778. doi: 10.1016/0092-8674(93)90496-d. [DOI] [PubMed] [Google Scholar]
  28. Matlashewski G. J., Tuck S., Pim D., Lamb P., Schneider J., Crawford L. V. Primary structure polymorphism at amino acid residue 72 of human p53. Mol Cell Biol. 1987 Feb;7(2):961–963. doi: 10.1128/mcb.7.2.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Matlashewski G., Banks L., Pim D., Crawford L. Analysis of human p53 proteins and mRNA levels in normal and transformed cells. Eur J Biochem. 1986 Feb 3;154(3):665–672. doi: 10.1111/j.1432-1033.1986.tb09449.x. [DOI] [PubMed] [Google Scholar]
  30. Matlashewski G., Lamb P., Pim D., Peacock J., Crawford L., Benchimol S. Isolation and characterization of a human p53 cDNA clone: expression of the human p53 gene. EMBO J. 1984 Dec 20;3(13):3257–3262. doi: 10.1002/j.1460-2075.1984.tb02287.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Melefors O., Hentze M. W. Translational regulation by mRNA/protein interactions in eukaryotic cells: ferritin and beyond. Bioessays. 1993 Feb;15(2):85–90. doi: 10.1002/bies.950150203. [DOI] [PubMed] [Google Scholar]
  32. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Milner J. A conformation hypothesis for the suppressor and promoter functions of p53 in cell growth control and in cancer. Proc Biol Sci. 1991 Aug 22;245(1313):139–145. doi: 10.1098/rspb.1991.0100. [DOI] [PubMed] [Google Scholar]
  34. Minden M. D., Buick R. N., McCulloch E. A. Separation of blast cell and T-lymphocyte progenitors in the blood of patients with acute myeloblastic leukemia. Blood. 1979 Jul;54(1):186–195. [PubMed] [Google Scholar]
  35. Moll U. M., LaQuaglia M., Bénard J., Riou G. Wild-type p53 protein undergoes cytoplasmic sequestration in undifferentiated neuroblastomas but not in differentiated tumors. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4407–4411. doi: 10.1073/pnas.92.10.4407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Moll U. M., Riou G., Levine A. J. Two distinct mechanisms alter p53 in breast cancer: mutation and nuclear exclusion. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7262–7266. doi: 10.1073/pnas.89.15.7262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Momand J., Zambetti G. P., Olson D. C., George D., Levine A. J. The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation. Cell. 1992 Jun 26;69(7):1237–1245. doi: 10.1016/0092-8674(92)90644-r. [DOI] [PubMed] [Google Scholar]
  38. Oliner J. D., Kinzler K. W., Meltzer P. S., George D. L., Vogelstein B. Amplification of a gene encoding a p53-associated protein in human sarcomas. Nature. 1992 Jul 2;358(6381):80–83. doi: 10.1038/358080a0. [DOI] [PubMed] [Google Scholar]
  39. Papayannopoulou T., Nakamoto B., Kurachi S., Tweeddale M., Messner H. Surface antigenic profile and globin phenotype of two new human erythroleukemia lines: characterization and interpretations. Blood. 1988 Sep;72(3):1029–1038. [PubMed] [Google Scholar]
  40. Pause A., Méthot N., Sonenberg N. The HRIGRXXR region of the DEAD box RNA helicase eukaryotic translation initiation factor 4A is required for RNA binding and ATP hydrolysis. Mol Cell Biol. 1993 Nov;13(11):6789–6798. doi: 10.1128/mcb.13.11.6789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Rogel A., Popliker M., Webb C. G., Oren M. p53 cellular tumor antigen: analysis of mRNA levels in normal adult tissues, embryos, and tumors. Mol Cell Biol. 1985 Oct;5(10):2851–2855. doi: 10.1128/mcb.5.10.2851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sasano H., Goukon Y., Nishihira T., Nagura H. In situ hybridization and immunohistochemistry of p53 tumor suppressor gene in human esophageal carcinoma. Am J Pathol. 1992 Sep;141(3):545–550. [PMC free article] [PubMed] [Google Scholar]
  43. Scheffner M., Werness B. A., Huibregtse J. M., Levine A. J., Howley P. M. The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53. Cell. 1990 Dec 21;63(6):1129–1136. doi: 10.1016/0092-8674(90)90409-8. [DOI] [PubMed] [Google Scholar]
  44. Slingerland J. M., Minden M. D., Benchimol S. Mutation of the p53 gene in human acute myelogenous leukemia. Blood. 1991 Apr 1;77(7):1500–1507. [PubMed] [Google Scholar]
  45. Smith L. J., McCulloch E. A., Benchimol S. Expression of the p53 oncogene in acute myeloblastic leukemia. J Exp Med. 1986 Sep 1;164(3):751–761. doi: 10.1084/jem.164.3.751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sugimoto K., Hirano N., Toyoshima H., Chiba S., Mano H., Takaku F., Yazaki Y., Hirai H. Mutations of the p53 gene in myelodysplastic syndrome (MDS) and MDS-derived leukemia. Blood. 1993 Jun 1;81(11):3022–3026. [PubMed] [Google Scholar]
  47. Sugimoto K., Toyoshima H., Sakai R., Miyagawa K., Hagiwara K., Hirai H., Ishikawa F., Takaku F. Mutations of the p53 gene in lymphoid leukemia. Blood. 1991 Mar 15;77(6):1153–1156. [PubMed] [Google Scholar]
  48. Torczynski R. M., Fuke M., Bollon A. P. Cloning and sequencing of a human 18S ribosomal RNA gene. DNA. 1985 Aug;4(4):283–291. doi: 10.1089/dna.1985.4.283. [DOI] [PubMed] [Google Scholar]
  49. Trecca D., Longo L., Biondi A., Cro L., Calori R., Grignani F., Maiolo A. T., Pelicci P. G., Neri A. Analysis of p53 gene mutations in acute myeloid leukemia. Am J Hematol. 1994 Aug;46(4):304–309. doi: 10.1002/ajh.2830460409. [DOI] [PubMed] [Google Scholar]
  50. Tuck S. P., Crawford L. Characterization of the human p53 gene promoter. Mol Cell Biol. 1989 May;9(5):2163–2172. doi: 10.1128/mcb.9.5.2163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Ueda H., Ullrich S. J., Gangemi J. D., Kappel C. A., Ngo L., Feitelson M. A., Jay G. Functional inactivation but not structural mutation of p53 causes liver cancer. Nat Genet. 1995 Jan;9(1):41–47. doi: 10.1038/ng0195-41. [DOI] [PubMed] [Google Scholar]
  52. Ullrich S. J., Mercer W. E., Appella E. Human wild-type p53 adopts a unique conformational and phosphorylation state in vivo during growth arrest of glioblastoma cells. Oncogene. 1992 Aug;7(8):1635–1643. [PubMed] [Google Scholar]
  53. Wang C., Curtis J. E., Minden M. D., McCulloch E. A. Expression of a retinoic acid receptor gene in myeloid leukemia cells. Leukemia. 1989 Apr;3(4):264–269. [PubMed] [Google Scholar]
  54. Wattel E., Preudhomme C., Hecquet B., Vanrumbeke M., Quesnel B., Dervite I., Morel P., Fenaux P. p53 mutations are associated with resistance to chemotherapy and short survival in hematologic malignancies. Blood. 1994 Nov 1;84(9):3148–3157. [PubMed] [Google Scholar]
  55. Wiggans R. G., Jacobson R. J., Fialkow P. J., Woolley P. V., 3rd, Macdonald J. S., Schein P. S. Probable clonal origin of acute myeloblastic leukemia following radiation and chemotherapy of colon cancer. Blood. 1978 Oct;52(4):659–663. [PubMed] [Google Scholar]
  56. Winship P. R. An improved method for directly sequencing PCR amplified material using dimethyl sulphoxide. Nucleic Acids Res. 1989 Feb 11;17(3):1266–1266. doi: 10.1093/nar/17.3.1266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Winter E., Yamamoto F., Almoguera C., Perucho M. A method to detect and characterize point mutations in transcribed genes: amplification and overexpression of the mutant c-Ki-ras allele in human tumor cells. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7575–7579. doi: 10.1073/pnas.82.22.7575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Zhan Q., Carrier F., Fornace A. J., Jr Induction of cellular p53 activity by DNA-damaging agents and growth arrest. Mol Cell Biol. 1993 Jul;13(7):4242–4250. doi: 10.1128/mcb.13.7.4242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Zhang W., Hu G., Estey E., Hester J., Deisseroth A. Altered conformation of the p53 protein in myeloid leukemia cells and mitogen-stimulated normal blood cells. Oncogene. 1992 Aug;7(8):1645–1647. [PubMed] [Google Scholar]
  60. Zhu Y. M., Bradbury D., Russell N. Expression of different conformations of p53 in the blast cells of acute myeloblastic leukaemia is related to in vitro growth characteristics. Br J Cancer. 1993 Nov;68(5):851–855. doi: 10.1038/bjc.1993.444. [DOI] [PMC free article] [PubMed] [Google Scholar]

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