The transcription factor Sp1 is implicated in the activation of G0/G1 phase genes. Modulation of Sp1 transcription activities may affect G1-S checkpoint, resulting in changes in cell proliferation. In this study, our results demonstrated that activated poly(ADP-ribose) polymerase 1 (PARP-1) promoted cell proliferation by inhibiting Sp1 signaling pathway. Cell proliferation and cell cycle assays demonstrated that PARP inhibitors or PARP-1 siRNA treatment significantly inhibited proliferation of hepatoma cells and induced G0/G1 cell cycle arrest in hepatoma cells, while overexpression of PARP-1 or PARP-1 activator treatment promoted cell cycle progression. Simultaneously, inhibition of PARP-1 enhanced the expression of Sp1-mediated checkpoint proteins, such as p21 and p27. In this study, we also showed that Sp1 was poly(ADP-ribosyl)ated by PARP-1 in hepatoma cells. Poly(ADP-ribosyl)ation suppressed Sp1 mediated transcription through preventing Sp1 binding to the Sp1 response element present in the promoters of target genes. Taken together, these data indicated that PARP-1 inhibition attenuated the poly(ADP-ribosyl)ation of Sp1 and significantly increased the expression of Sp1 target genes, resulting in G0/G1 cell cycle arrest and the decreased proliferative ability of the hepatoma cells.
References
[1]
Kadonaga JT, Carner KR, Masiarz FR, Tjian R (1987) Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain. Cell 51: 1079–1090.
[2]
Black AR, Black JD, Azizkhan-Clifford J (2001) Sp1 and kruppel-like factor family of transcription factors in cell growth regulation and cancer. J Cell Physiol 188: 143–160.
[3]
Song J, Ugai H, Ogawa K, Wang Y, Sarai A, et al. (2001) Two consecutive zinc fingers in Sp1 and in MAZ are essential for interactions with cis-elements. J Biol Chem 276: 30429–30434.
[4]
Tapias A, Ciudad CJ, Roninson IB, Noe V (2008) Regulation of Sp1 by cell cycle related proteins. Cell Cycle 7: 2856–2867.
[5]
Feng XH, Lin X, Derynck R (2000) Smad2, Smad3 and Smad4 cooperate with Sp1 to induce p15(Ink4B) transcription in response to TGF-beta. EMBO J 19: 5178–5193.
[6]
Wu J, Xue L, Weng M, Sun Y, Zhang Z, et al. (2007) Sp1 is essential for p16 expression in human diploid fibroblasts during senescence. PLoS One 2: e164.
[7]
Deniaud E, Baguet J, Chalard R, Blanquier B, Brinza L, et al. (2009) Overexpression of transcription factor Sp1 leads to gene expression perturbations and cell cycle inhibition. PLoS One 4: e7035.
[8]
Biggs JR, Kudlow JE, Kraft AS (1996) The role of the transcription factor Sp1 in regulating the expression of the WAF1/CIP1 gene in U937 leukemic cells. J Biol Chem 271: 901–906.
[9]
Chen F, Kim E, Wang CC, Harrison LE (2005) Ciglitazone-induced p27 gene transcriptional activity is mediated through Sp1 and is negatively regulated by the MAPK signaling pathway. Cell Signal 17: 1572–1577.
[10]
Gartel AL, Goufman E, Najmabadi F, Tyner AL (2000) Sp1 and Sp3 activate p21 (WAF1/CIP1) gene transcription in the Caco-2 colon adenocarcinoma cell line. Oncogene 19: 5182–5188.
[11]
Abdelrahim M, Smith R 3rd, Burghardt R, Safe S (2004) Role of Sp proteins in regulation of vascular endothelial growth factor expression and proliferation of pancreatic cancer cells. Cancer Res 64: 6740–6749.
[12]
Mertens-Talcott SU, Chintharlapalli S, Li X, Safe S (2007) The oncogenic microRNA-27a targets genes that regulate specificity protein transcription factors and the G2-M checkpoint in MDA-MB-231 breast cancer cells. Cancer Res 67: 11001–11011.
[13]
Wang YT, Chuang JY, Shen MR, Yang WB, Chang WC, et al. (2008) Sumoylation of specificity protein 1 augments its degradation by changing the localization and increasing the specificity protein 1 proteolytic process. J Mol Biol 380: 869–885.
[14]
Chuang JY, Wu CH, Lai MD, Chang WC, Hung JJ (2009) Overexpression of Sp1 leads to p53-dependent apoptosis in cancer cells. Int J Cancer 125: 2066–2076.
[15]
Li L, Davie JR (2010) The role of Sp1 and Sp3 in normal and cancer cell biology. Ann Anat 192: 275–283.
[16]
Wang L, Wei D, Huang S, Peng Z, Le X, et al. (2003) Transcription factor Sp1 expression is a significant predictor of survival in human gastric cancer. Clin Cancer Res 9: 6371–6380.
[17]
Hosoi Y, Watanabe T, Nakagawa K, Matsumoto Y, Enomoto A, et al. (2004) Up-regulation of DNA-dependent protein kinase activity and Sp1 in colorectal cancer. Int J Oncol 25: 461–468.
[18]
Schreiber V, Dantzer F, Ame JC, de Murcia G (2006) Poly(ADP-ribose): novel functions for an old molecule. Nat Rev Mol Cell Biol 7: 517–528.
[19]
D'Amours D, Desnoyers S, D'Silva I, Poirier GG (1999) Poly(ADP-ribosyl)ation reactions in the regulation of nuclear functions. Biochem J 342 (Pt 2): 249–268.
[20]
Prasad R, Lavrik OI, Kim SJ, Kedar P, Yang XP, et al. (2001) DNA polymerase beta -mediated long patch base excision repair. Poly(ADP-ribose)polymerase-1 stimulates strand displacement DNA synthesis. J Biol Chem 276: 32411–32414.
[21]
Pleschke JM, Kleczkowska HE, Strohm M, Althaus FR (2000) Poly(ADP-ribose) binds to specific domains in DNA damage checkpoint proteins. J Biol Chem 275: 40974–40980.
[22]
Carbone M, Rossi MN, Cavaldesi M, Notari A, Amati P, et al. (2008) Poly(ADP-ribosyl)ation is implicated in the G0–G1 transition of resting cells. Oncogene 27: 6083–6092.
[23]
Augustin A, Spenlehauer C, Dumond H, Menissier-De Murcia J, Piel M, et al. (2003) PARP-3 localizes preferentially to the daughter centriole and interferes with the G1/S cell cycle progression. J Cell Sci 116: 1551–1562.
[24]
Simbulan-Rosenthal CM, Rosenthal DS, Luo R, Samara R, Espinoza LA, et al. (2003) PARP-1 binds E2F-1 independently of its DNA binding and catalytic domains, and acts as a novel coactivator of E2F-1-mediated transcription during re-entry of quiescent cells into S phase. Oncogene 22: 8460–8471.
[25]
Anderson MG, Scoggin KE, Simbulan-Rosenthal CM, Steadman JA (2000) Identification of poly(ADP-ribose) polymerase as a transcriptional coactivator of the human T-cell leukemia virus type 1 Tax protein. J Virol 74: 2169–2177.
[26]
Grinstein E, Jundt F, Weinert I, Wernet P, Royer HD (2002) Sp1 as G1 cell cycle phase specific transcription factor in epithelial cells. Oncogene 21: 1485–1492.
[27]
Kim MY, Zhang T, Kraus WL (2005) Poly(ADP-ribosyl)ation by PARP-1: 'PAR-laying' NAD+ into a nuclear signal. Genes Dev 19: 1951–1967.
[28]
Zaniolo K, Desnoyers S, Leclerc S, Guerin SL (2007) Regulation of poly(ADP-ribose) polymerase-1 (PARP-1) gene expression through the post-translational modification of Sp1: a nuclear target protein of PARP-1. BMC Mol Biol 8: 96.
[29]
Huang D, Wang Y, Yang C, Liao Y, Huang K (2009) Angiotensin II promotes poly(ADP-ribosyl)ation of c-Jun/c-Fos in cardiac fibroblasts. J Mol Cell Cardiol 46: 25–32.
[30]
Milutinovic S, Brown SE, Zhuang Q, Szyf M (2004) DNA methyltransferase 1 knock down induces gene expression by a mechanism independent of DNA methylation and histone deacetylation. J Biol Chem 279: 27915–27927.
[31]
Shimo T, Kurebayashi J, Kanomata N, Yamashita T, Kozuka Y, et al.. (2012) Antitumor and anticancer stem cell activity of a poly ADP-ribose polymerase inhibitor olaparib in breast cancer cells. Breast Cancer.
[32]
Dynan WS, Saffer JD, Lee WS, Tjian R (1985) Transcription factor Sp1 recognizes promoter sequences from the monkey genome that are simian virus 40 promoter. Proc Natl Acad Sci U S A 82: 4915–4919.
[33]
Jackson SP, Tjian R (1988) O-glycosylation of eukaryotic transcription factors: implications for mechanisms of transcriptional regulation. Cell 55: 125–133.
[34]
Chuang JY, Wang YT, Yeh SH, Liu YW, Chang WC, et al. (2008) Phosphorylation by c-Jun NH2-terminal kinase 1 regulates the stability of transcription factor Sp1 during mitosis. Mol Biol Cell 19: 1139–1151.
[35]
Tan NY, Khachigian LM (2009) Sp1 phosphorylation and its regulation of gene transcription. Mol Cell Biol 29: 2483–2488.
[36]
Wierstra I (2008) Sp1: emerging roles–beyond constitutive activation of TATA-less housekeeping genes. Biochem Biophys Res Commun 372: 1–13.
[37]
Shieh WM, Ame JC, Wilson MV, Wang ZQ, Koh DW, et al. (1998) Poly(ADP-ribose) polymerase null mouse cells synthesize ADP-ribose polymers. J Biol Chem 273: 30069–30072.
[38]
Oei SL, Griesenbeck J, Schweiger M, Ziegler M (1998) Regulation of RNA polymerase II-dependent transcription by poly(ADP-ribosyl)ation of transcription factors. J Biol Chem 273: 31644–31647.
[39]
Huang D, Yang C, Wang Y, Liao Y, Huang K (2009) PARP-1 suppresses adiponectin expression through poly(ADP-ribosyl)ation of PPAR gamma in cardiac fibroblasts. Cardiovasc Res 81: 98–107.
[40]
Curtin NJ (2005) PARP inhibitors for cancer therapy. Expert Rev Mol Med 7: 1–20.
[41]
Kirby CA, Cheung A, Fazal A, Shultz MD, Stams T (2012) Structure of human tankyrase 1 in complex with small-molecule inhibitors PJ34 and XAV939. Acta Crystallogr Sect F Struct Biol Cryst Commun 68: 115–118.
[42]
Simonin F, Menissier-de Murcia J, Poch O, Muller S, Gradwohl G, et al. (1990) Expression and site-directed mutagenesis of the catalytic domain of human poly(ADP-ribose)polymerase in Escherichia coli. Lysine 893 is critical for activity. J Biol Chem 265: 19249–19256.
[43]
Salic A, Mitchison TJ (2008) A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc Natl Acad Sci U S A 105: 2415–2420.
[44]
Zeng C, Luo Y, Asico LD, Hopfer U, Eisner GM, et al. (2003) Perturbation of D1 dopamine and AT1 receptor interaction in spontaneously hypertensive rats. Hypertension 42: 787–792.
[45]
Xu W, Cho H, Evans RM (2003) Acetylation and methylation in nuclear receptor gene activation. Methods Enzymol 364: 205–223.
[46]
Wilson AJ, Byun DS, Nasser S, Murray LB, Ayyanar K, et al. (2008) HDAC4 promotes growth of colon cancer cells via repression of p21. Mol Biol Cell 19: 4062–4075.
