Abstract
Protein phosphatase 1 (PP1) plays important roles in many different aspects of cellular activities including cell cycle control. One important function of PP1 is to activate the retinoblastoma protein pRB. Here we show that pRB is one of PP1's downstream targets during apoptosis. When HL-60 cells synchronized at the G1/S boundary were treated with pro-apoptotic cytosine arabinoside (araC), PP1α protein increased twofold and PP1 activity about 30% within 1 h. This was followed by pRB dephosphorylation, pRB cleavage by caspases, DNA fragmentation, the appearance of cells with <2n DNA content and finally, dying and dead cells. In vitro, pRB was protected from caspase-3 digestion by prior Cdk-mediated phosphorylation, whereas PP1α converted phospho-pRB into an efficient substrate for caspase-3. Introduction of active PP1α into HL-60 cells by electroporation was sufficient to induce characteristics of apoptosis. Similarly, araC-resistant cells, normally unable to die in response to araC, initiated apoptosis when electroporated with active PP1α. This was also accompanied by pRB cleavage. In contrast, introduction of inhibitor-2 delayed the onset of araC-induced apoptosis, whereas concomitant introduction of PP1α and inhibitor-2 completely prevented PP1α-induced apoptosis. These results suggest that dephosphorylation of key proteins by PP1α may be crucial for the initiation of apoptosis and further support the concept of PP1 serving as a potential target for anti-cancer therapy.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Alberts AS, Thorburn AM, Shenolikar S, Mumby MC, Feramisco JR . 1993 Proc. Natl. Acad. Sci. USA 90: 388–392
Axton JM, Dómbradi V, Cohen PTW, Glover DM . 1990 Cell 63: 33–46
Ayllón V, Martínez-A C, García A, Cayla X, Rebollo A . 2000 EMBO J. 19: 2237–2246
Baxter GD, Lavin MF . 1992 J. Immunol. 148: 1949–1954
Berndt N . 1995 Adv. Prot. Phosphatases 9: 63–86
Berndt N . 1999 Front. Biosci. 4: d22–d42
Berndt N . 2000 Emerg. Ther. Targets 4: 581–608
Berndt N, Dohadwala M, Liu CWY . 1997 Curr. Biol. 7: 375–386
Berry DE, Lu Y, Schmidt B, Fallon PG, O'Connell C, Hu S-X, Xu H-J, Blanck G . 1996 Oncogene 12: 1809–1819
Bøe R, Gjertsen BT, Vintermyr OK, Houge G, Lanotte M, Døskeland SO . 1991 Exp. Cell Res. 195: 237–246
Booher RN, Beach D . 1989 Cell 57: 1009–1016
Bradford MM . 1976 Anal. Biochem. 72: 248–254
Cardone MH, Roy N, Stennicke HR, Salvesen GS, Franke TF, Stanbridge EJ, Frisch S, Reed JC . 1998 Science 282: 1318–1321
Chen W, Otterson GA, Lipkowitz S, Khleif SN, Coxon AB, Kaye FJ . 1997 Oncogene 14: 1243–1248
Cheng AY, Dean NM, Honkanen RE . 2000 J. Biol. Chem. 275: 1846–1854
Clarke AR, Maandag ER, Van Roon M, Van der Lugt NMT, Van der Valk M, Hooper ML, Berns A, Te Riele H . 1992 Nature 359: 328–330
Damer CK, Partridge J, Pearson WR, Haystead TAJ . 1998 J. Biol. Chem. 273: 24396–24405
Dohadwala M, Berndt N . 1998 Protein Phosphatase Protocols Vol 93. Ludlow JW (ed) Humana Press: Totowa, NJ pp 191–199
Dohadwala M, Da Cruz e Silva EF, Hall FL, Williams RT, Carbonaro-Hall DA, Nairn AC, Greengard P, Berndt N . 1994 Proc. Natl. Acad. Sci. USA 91: 6408–6412
Dou QP, An B, Will PL . 1995 Proc. Natl. Acad. Sci. USA 92: 9019–9023
Downward J . 1998 Curr. Opin. Cell Biol. 10: 262–267
Dunigan DD, Madlener JC . 1995 Virology 207: 460–466
Durfee T, Becherer K, Chen P-L, Yeh S-H, Yang Y, Kilburn AE, Lee W-H, Elledge SJ . 1993 Genes Dev. 7: 555–569
Fan G, Ma X, Kren BT, Steer CJ . 1996 Oncogene 12: 1909–1919
Fernandez A, Brautigan DL, Lamb NJC . 1992 J. Cell Biol. 116: 1421–1430
Green DR . 1998 Cell 94: 695–698
Haas-Kogan DA, Kogan SC, Levi D, Dazin P, T'Ang A, Fung Y-KT, Israel MA . 1995 EMBO J. 14: 461–472
Hotz MA, Gong J, Traganos F, Darzynkiewicz Z . 1994 Cytometry 15: 237–244
Inomata M, Saijo N, Kawashima K, Kaneko A, Fujiwara Y, Kunikane H, Tanaka Y . 1995 J. Cancer Res. Clin. Oncol. 121: 729–738
Jacks T, Fazeli A, Schmitt EM, Bronson RT, Goodell MA, Weinberg RA . 1992 Nature 359: 295–300
Jänicke RU, Walker PA, Lin XY, Porter AG . 1996 EMBO J. 15: 6969–6978
Knudsen KE, Weber E, Arden KC, Cavenee WK, Feramisco JR, Knudsen ES . 1999 Oncogene 18: 5239–5245
Kwon Y-G, Lee S-Y, Choi Y, Greengard P, Nairn AC . 1997 Proc. Natl. Acad. Sci. USA 94: 2168–2173
Laemmli UK . 1970 Nature 227: 680–685
Lazebnik YA, Kaufmann SH, Desnoyer S, Poirier GG, Earnshaw WC . 1994 Nature 371: 346–347
Lee EYHP, Chang C-Y, Hu N, Wang Y-CJ, Lai C-C, Herrup K, Lee W-H, Bradley A . 1992 Nature 359: 288–294
Liu CWY, Wang R-H, Dohadwala M, Schönthal AH, Villa-Moruzzi E, Berndt N . 1999 J. Biol. Chem. 274: 29470–29475
Ludlow JW, Glendening CL, Livingston DM, DeCaprio JA . 1993 Mol. Cell. Biol. 13: 367–372
Martin-Aragon S, Mukherjee SK, Taylor BJ, Ivy SP, Fu CH, Ardi VC, Avramis VI . 2000 Anticancer Res. 20: 139–150
Martin SJ, Green DR . 1995 Cell 82: 349–352
Mihara K, Cao X-R, Yen A, Chandler S, Driscoll B, Murphree AL, T'Ang A, Fung Y-KT . 1989 Science 246: 1300–1303
Morana SJ, Wolf CM, Li J, Reynolds JE, Brown MK, Eastman A . 1996 J. Biol. Chem. 271: 18263–18271
Morimoto Y, Ohba T, Kobayashi S, Haneji T . 1997 Exp. Cell Res. 230: 181–186
Ohkura H, Kinoshita N, Miyatani S, Toda T, Yanagida M . 1989 Cell 57: 997–1007
Puntoni F, Villa-Moruzzi E . 1999 Biochem. Biophys. Res. Commun. 266: 279–283
Schönthal AH, Feramisco JR . 1993 Oncogene 8: 433–441
Shibasaki F, McKeon F . 1995 J. Cell Biol. 131: 735–743
Spector DL, Goldman RD, Leinwand LA . 1998 Cells–A Laboratory Manual Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY
Takemura M, Kitagawa T, Izuta S, Wasa J, Takai A, Akiyama T, Yoshida S . 1997 Oncogene 15: 2483–2492
Tan X, Wang JYJ . 1998 Trends Cell Biol. 8: 116–120
Taylor BK, Stoops TD, Everett AD . 2000 Am. J. Physiol. 278: L1062–L1070
Wang HG, Pathan N, Ethell IM, Krajewski S, Yamaguchi Y, Shibasaki F, McKeon F, Bobo T, Franke TF, Reed JC . 1999 Science 284: 339–343
Yamano H, Ishii K, Yanagida M . 1994 EMBO J. 13: 5310–5318
Yi H-K, Fujimura Y, Ouchida M, Prasad DDK, Rao VN, Reddy SP . 1997 Oncogene 14: 1259–1268
Zheng B, Woo CF, Kuo JF . 1991 J. Biol. Chem. 266: 10031–10034
Acknowledgements
This work was supported in part by NIH grant CA54167 (to N Berndt). We wish to thank Balwant S Khatra, Charles J Sherr, David O Morgan, and Emma Villa-Moruzzi for their kind sharing of vectors and antibodies.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, RH., Liu, C., Avramis, V. et al. Protein phosphatase 1α-mediated stimulation of apoptosis is associated with dephosphorylation of the retinoblastoma protein. Oncogene 20, 6111–6122 (2001). https://doi.org/10.1038/sj.onc.1204829
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1204829
Keywords
This article is cited by
-
Nuclear Protein Phosphatase 1 α (PP1A) Expression is Associated with Poor Prognosis in p53 Expressing Glioblastomas
Pathology & Oncology Research (2016)
-
Molecular biology of cantharidin in cancer cells
Chinese Medicine (2007)
-
Altered structure and deregulated expression of the tumor suppressor gene retinoblastoma (RB1) in human brain tumors
Molecular and Cellular Biochemistry (2007)
-
Downregulation of Fer induces PP1 activation and cell-cycle arrest in malignant cells
Oncogene (2006)
-
DARPP-32 expression arises after a phase of dysplasia in oesophageal squamous cell carcinoma
British Journal of Cancer (2004)