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PMID: 9895308 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

p21WAF1 expression by an activator of protein kinase C is regulated mainly at the post-transcriptional level in cells lacking p53: important role of RNA stabilization.

The Biochemical journal ·Vol. 337 ( Pt 3) ·1999-02-01 ·Pages 607-16

Akashi M, Osawa Y, Koeffler HP, Hachiya M

Abstract

p21(WAF1) inhibits cyclin-cyclin-dependent kinase (Cdk) complexes, causing cell cycle arrest. p21(WAF1) contains p53-binding sites in its promoter and expression of p21(WAF1) is induced by functional p53. In the present work, we have studied the role of protein kinase C (PKC) in the induction of p21(WAF1) and show that induction of p21(WAF1) expression can occur by activation of PKC in cells having no p53. Human ovarian carcinoma cells, SKOV-3, lack p53 protein and PMA, a potent activator of PKC, did not induce p53. PMA increased the expression of p21(WAF1) mRNA both in these cells and in other cells which do not contain p53 (THP-1 and U937). Treatment of human embryonic fibroblasts, WI38, with PMA also induced the accumulation of p21(WAF1) without affecting p53 levels. However, PMA did not increase levels of p21(WAF1) mRNA in cells where either the PKC or the mitogen-activated protein kinase pathway was blocked. Furthermore, treatment of cells with various phorbol ester derivatives which activate PKC resulted in the induction of p21(WAF1) in SKOV-3 cells. In contrast, phorbol esters which do not activate PKC failed to induce p21(WAF1) expression. PMA increased the transcriptional rate of p21(WAF1) and activated the transcription of a luciferase reporter gene, controlled by the p21 promoter, in SKOV-3 cells with or without a p53 consensus-binding sequence. By contrast, PMA markedly stabilized p21(WAF1) mRNA; the half-life (t1/2) of p21(WAF1) in PMA-treated cells was >8 h compared with <1 h in untreated cells. These findings provide evidence that the PKC pathway induces expression of p21(WAF1) independently of p53. Our present study also suggests that the accumulation of p21(WAF1) transcripts by PMA occurs mainly at post-transcriptional level.

MeSH Terms
Calcium-Calmodulin-Dependent Protein Kinases/antagonists & inhibitors Cell Cycle/drug effects Cell Line Cyclin-Dependent Kinase Inhibitor p21 Cyclins/genetics,metabolism Enzyme Activation Gene Expression Regulation Humans Phorbol Esters/pharmacology Protein Kinase C/metabolism RNA/metabolism RNA, Messenger/metabolism Transcription, Genetic Tumor Suppressor Protein p53/deficiency
Chemicals
CDKN1A protein, human Cyclin-Dependent Kinase Inhibitor p21 Cyclins Phorbol Esters RNA, Messenger Tumor Suppressor Protein p53 RNA Protein Kinase C Calcium-Calmodulin-Dependent Protein Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Akashi M
Division of Radiation Health, National Institute of Radiological Sciences, Chiba, 263-8555 Japan. [email protected]
Osawa Y
Koeffler H P
Hachiya M
References (66)
66 references, click to expand
  1. Definition of a consensus binding site for p53.
    Nat Genet. 1992 Apr;1(1):45-9 PMID: 1301998
  2. Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C.
    Science. 1992 Oct 23;258(5082):607-14 PMID: 1411571
  3. Irradiation increases levels of GM-CSF through RNA stabilization which requires an AU-rich region in cancer cells.
    Biochem Biophys Res Commun. 1992 Dec 15;189(2):986-93 PMID: 1472071
  4. p53 function and dysfunction.
    Cell. 1992 Aug 21;70(4):523-6 PMID: 1505019
  5. Frequent mutations in the p53 gene in human myeloid leukemia cell lines.
    Blood. 1992 May 1;79(9):2378-83 PMID: 1571549
  6. Abnormal structure and expression of the p53 gene in human ovarian carcinoma cell lines.
    Cancer Res. 1992 Aug 1;52(15):4196-9 PMID: 1638534
  7. Levels of p53 protein increase with maturation in human hematopoietic cells.
    Cancer Res. 1991 Aug 15;51(16):4279-86 PMID: 1868448
  8. Regulation of interleukin 3 mRNA expression in mast cells occurs at the posttranscriptional level and is mediated by calcium ions.
    Proc Natl Acad Sci U S A. 1990 Jan;87(2):777-81 PMID: 2105489
  9. Cell cycle control in eukaryotes: molecular mechanisms of cdc2 activation.
    Trends Biochem Sci. 1990 Oct;15(10):378-83 PMID: 2147521
  10. Abnormal structure and expression of p53 gene in human hepatocellular carcinoma.
    Proc Natl Acad Sci U S A. 1990 Mar;87(5):1973-7 PMID: 2155427
  11. Cell proliferation in carcinogenesis.
    Science. 1990 Aug 31;249(4972):1007-11 PMID: 2204108
  12. Monoclonal antibody analysis of p53 expression in normal and transformed cells.
    J Virol. 1986 Aug;59(2):444-52 PMID: 2426467
  13. Regulation of lymphokine messenger RNA stability by a surface-mediated T cell activation pathway.
    Science. 1989 Apr 21;244(4902):339-43 PMID: 2540528
  14. Lymphotoxin: stimulation and regulation of colony-stimulating factors in fibroblasts.
    Blood. 1989 Nov 15;74(7):2383-90 PMID: 2679916
  15. Complementary DNA for human T-cell cyclophilin.
    EMBO J. 1987 Apr;6(4):947-50 PMID: 3297675
  16. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation.
    Cell. 1986 Aug 29;46(5):659-67 PMID: 3488815
  17. Growth factor-stimulated protein phosphorylation in 3T3-L1 cells. Evidence for protein kinase C-dependent and -independent pathways.
    J Biol Chem. 1985 Oct 25;260(24):13304-15 PMID: 3902816
  18. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
    Anal Biochem. 1983 Jul 1;132(1):6-13 PMID: 6312838
  19. Extreme instability of myc mRNA in normal and transformed human cells.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):7046-50 PMID: 6594679
  20. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters.
    J Biol Chem. 1982 Jul 10;257(13):7847-51 PMID: 7085651
  21. mRNA stability in mammalian cells.
    Microbiol Rev. 1995 Sep;59(3):423-50 PMID: 7565413
  22. Inhibition of cyclin-dependent kinases by p21.
    Mol Biol Cell. 1995 Apr;6(4):387-400 PMID: 7626805
  23. Irradiation induces WAF1 expression through a p53-independent pathway in KG-1 cells.
    J Biol Chem. 1995 Aug 11;270(32):19181-7 PMID: 7642586
  24. A synthetic inhibitor of the mitogen-activated protein kinase cascade.
    Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7686-9 PMID: 7644477
  25. Cloning of p57KIP2, a cyclin-dependent kinase inhibitor with unique domain structure and tissue distribution.
    Genes Dev. 1995 Mar 15;9(6):639-49 PMID: 7729683
  26. Inhibitors of mammalian G1 cyclin-dependent kinases.
    Genes Dev. 1995 May 15;9(10):1149-63 PMID: 7758941
  27. p53-dependent and independent expression of p21 during cell growth, differentiation, and DNA damage.
    Genes Dev. 1995 Apr 15;9(8):935-44 PMID: 7774811
  28. How MAP kinases are regulated.
    J Biol Chem. 1995 Jun 23;270(25):14843-6 PMID: 7797459
  29. Mutant p53 proteins behave in a dominant, negative fashion in vivo.
    Anticancer Res. 1994 Sep-Oct;14(5A):1853-9 PMID: 7847818
  30. Correlation of terminal cell cycle arrest of skeletal muscle with induction of p21 by MyoD.
    Science. 1995 Feb 17;267(5200):1018-21 PMID: 7863327
  31. Gene transfer with a series of lipophilic DNA-binding molecules.
    Bioconjug Chem. 1994 Nov-Dec;5(6):647-54 PMID: 7873668
  32. p53 in hematologic malignancies.
    Blood. 1994 Oct 15;84(8):2412-21 PMID: 7919360
  33. Differential effects by the p21 CDK inhibitor on PCNA-dependent DNA replication and repair.
    Nature. 1994 Oct 6;371(6497):534-7 PMID: 7935768
  34. p21-containing cyclin kinases exist in both active and inactive states.
    Genes Dev. 1994 Aug 1;8(15):1750-8 PMID: 7958854
  35. Mechanisms of regulation of WAF1/Cip1 gene expression in human breast carcinoma: role of p53-dependent and independent signal transduction pathways.
    Oncogene. 1994 Dec;9(12):3407-15 PMID: 7970699
  36. Induction of WAF1/CIP1 by a p53-independent pathway.
    Cancer Res. 1994 Jul 1;54(13):3391-5 PMID: 8012956
  37. Cloning of p27Kip1, a cyclin-dependent kinase inhibitor and a potential mediator of extracellular antimitogenic signals.
    Cell. 1994 Jul 15;78(1):59-66 PMID: 8033212
  38. Effects of c-myc expression on cell cycle progression.
    Mol Cell Biol. 1994 Sep;14(9):5748-55 PMID: 8065309
  39. WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis.
    Cancer Res. 1994 Mar 1;54(5):1169-74 PMID: 8118801
  40. p53-dependent inhibition of cyclin-dependent kinase activities in human fibroblasts during radiation-induced G1 arrest.
    Cell. 1994 Mar 25;76(6):1013-23 PMID: 8137420
  41. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases.
    Cell. 1993 Nov 19;75(4):805-16 PMID: 8242751
  42. WAF1, a potential mediator of p53 tumor suppression.
    Cell. 1993 Nov 19;75(4):817-25 PMID: 8242752
  43. p21 is a universal inhibitor of cyclin kinases.
    Nature. 1993 Dec 16;366(6456):701-4 PMID: 8259214
  44. The selective protein kinase C inhibitor GF 109203X inhibits phorbol ester-induced morphological and functional differentiation of SH-SY5Y human neuroblastoma cells.
    Biochem Biophys Res Commun. 1993 Dec 30;197(3):1185-93 PMID: 8280132
  45. Posttranscriptional down-regulation of ras oncogene expression by inhibitors of cellular glutathione.
    Mol Cell Biol. 1993 Jul;13(7):4416-22 PMID: 8321241
  46. Protein kinase C alpha activates RAF-1 by direct phosphorylation.
    Nature. 1993 Jul 15;364(6434):249-52 PMID: 8321321
  47. Ca2+ ionophore A23187-dependent stabilization of granulocyte-macrophage colony-stimulating factor messenger RNA in murine thymoma EL-4 cells is mediated through two distinct regions in the 3'-untranslated region.
    J Immunol. 1993 May 15;150(10):4386-94 PMID: 8482841
  48. Mammalian G1 cyclins.
    Cell. 1993 Jun 18;73(6):1059-65 PMID: 8513492
  49. Regulation of p21WAF1/CIP1 expression through mitogen-activated protein kinase signaling pathway.
    Cancer Res. 1996 Jan 1;56(1):31-5 PMID: 8548769
  50. The role of the transcription factor Sp1 in regulating the expression of the WAF1/CIP1 gene in U937 leukemic cells.
    J Biol Chem. 1996 Jan 12;271(2):901-6 PMID: 8557703
  51. Transcriptional activation of the Cdk inhibitor p21 by vitamin D3 leads to the induced differentiation of the myelomonocytic cell line U937.
    Genes Dev. 1996 Jan 15;10(2):142-53 PMID: 8566748
  52. Tumor necrosis factor alpha: posttranscriptional stabilization of WAF1 mRNA in p53-deficient human leukemic cells.
    J Cell Physiol. 1996 Mar;166(3):568-76 PMID: 8600160
  53. Cell growth arrest and induction of cyclin-dependent kinase inhibitor p21 WAF1/CIP1 mediated by STAT1.
    Science. 1996 May 3;272(5262):719-22 PMID: 8614832
  54. Posttranscriptional regulation of gene expression in liver regeneration: role of mRNA stability.
    FASEB J. 1996 Apr;10(5):559-73 PMID: 8621056
  55. Phorbol ester inhibits the phosphorylation of the retinoblastoma protein without suppressing cyclin D-associated kinase in vascular smooth muscle cells.
    J Biol Chem. 1996 Apr 5;271(14):8345-51 PMID: 8626531
  56. The cell cycle-coupled expression of topoisomerase IIalpha during S phase is regulated by mRNA stability and is disrupted by heat shock or ionizing radiation.
    Mol Cell Biol. 1996 Apr;16(4):1500-8 PMID: 8657123
  57. Novel form of p21(WAF1/CIP1/SDI1) protein in phorbol ester-induced G2/M arrest.
    J Biol Chem. 1996 Nov 22;271(47):29556-60 PMID: 8939883
  58. Ubiquitination of p53 and p21 is differentially affected by ionizing and UV radiation.
    Mol Cell Biol. 1997 Jan;17(1):355-63 PMID: 8972216
  59. Signal transducer and activator of transcription-3 (STAT3) is constitutively activated in normal, self-renewing B-1 cells but only inducibly expressed in conventional B lymphocytes.
    J Exp Med. 1997 Mar 17;185(6):1035-42 PMID: 9091577
  60. Redox-mediated regulation of p21(waf1/cip1) expression involves a post-transcriptional mechanism and activation of the mitogen-activated protein kinase pathway.
    Eur J Biochem. 1997 May 1;245(3):730-7 PMID: 9183012
  61. Lysophosphatidic acid inhibits epidermal-growth-factor-induced Stat1 signaling in human epidermoid carcinoma A431 cells.
    Biochem Biophys Res Commun. 1997 Nov 26;240(3):856-61 PMID: 9398658
  62. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  63. Antiproliferative effect of 1alpha,25-dihydroxyvitamin D3 in human prostate cancer cell line LNCaP involves reduction of cyclin-dependent kinase 2 activity and persistent G1 accumulation.
    Endocrinology. 1998 Mar;139(3):1197-207 PMID: 9492054
  64. The role of protein stability in the cell cycle and cancer.
    Biochim Biophys Acta. 1998 Apr 17;1377(2):M61-70 PMID: 9606978
  65. The JAK-STAT pathway: signal transduction involved in proliferation, differentiation and transformation.
    Leuk Lymphoma. 1998 Feb;28(5-6):459-67 PMID: 9613975
  66. Induction of cell death by stimulation of protein kinase C in human epithelial cells expressing a mutant ras oncogene: a potential therapeutic target.
    Br J Cancer. 1998 Sep;78(5):641-51 PMID: 9744505
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
Email
[email protected]
Published
1999-02-01
Pages
607-16
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1220016
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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