This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Adnane, J.
Right arrow Articles by Sebti, S. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Adnane, J.
Right arrow Articles by Sebti, S. M.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, December 1998, p. 6962-6970, Vol. 18, No. 12
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

p21WAF1/CIP1 Is Upregulated by the Geranylgeranyltransferase I Inhibitor GGTI-298 through a Transforming Growth Factor beta - and Sp1-Responsive Element: Involvement of the Small GTPase RhoA

Jalila Adnane,1 Francisco A. Bizouarn,1 Yimin Qian,2 Andrew D. Hamilton,2 and Saïd M. Sebti1,*

Drug Discovery Program, H. Lee Moffitt Cancer Center, and Department of Biochemistry and Molecular Biology, University of South Florida, Tampa, Florida 33612,1 and Department of Chemistry, Yale University, New Haven, Connecticut2

Received 23 February 1998/Returned for modification 15 May 1998/Accepted 26 August 1998

We have recently reported that the geranylgeranyltransferase I inhibitor GGTI-298 arrests human tumor cells at the G1 phase of the cell cycle and increases the protein and RNA levels of the cyclin-dependent kinase inhibitor p21WAF1/CIP1. Here, we show that GGTI-298 acts at the transcriptional level to induce p21WAF1/CIP1 in a human pancreatic carcinoma cell line, Panc-1. This upregulation of p21WAF1/CIP1 promoter was selective, since GGTI-298 inhibited serum responsive element- and E2F-mediated transcription. A functional analysis of the p21WAF1/CIP1 promoter showed that a GC-rich region located between positions -83 and -74, which contains a transforming growth factor beta -responsive element and one Sp1-binding site, is sufficient for the upregulation of p21WAF1/CIP1 promoter by GGTI-298. Electrophoretic mobility shift assays showed a small increase in the amount of DNA-bound Sp1-Sp3 complexes. Furthermore, the analysis of Sp1 transcriptional activity in GGTI-298-treated cells by using GAL4-Sp1 chimera or Sp1-chloramphenicol acetyltransferase reporter revealed a significant increase in Sp1-mediated transcription. Moreover, GGTI-298 treatment also resulted in increased Sp1 and Sp3 phosphorylation. These results suggest that GGTI-298-mediated upregulation of p21WAF1/CIP1 involves both an increase in the amount of DNA-bound Sp1-Sp3 and enhancement of Sp1 transcriptional activity. To identify the geranylgeranylated protein(s) involved in p21WAF1/CIP1 transcriptional activation, we analyzed the effects of the small GTPases Rac1 and RhoA on p21WAF1/CIP1 promoter activity. The dominant negative mutant of RhoA, but not Rac1, was able to activate p21WAF1/CIP1. In contrast, constitutively active RhoA repressed p21WAF1/CIP1. Accordingly, the ADP-ribosyl transferase C3, which specifically inhibits Rho proteins, enhanced the activity of p21WAF1/CIP1. Taken together, these results suggest that one mechanism by which GGTI-298 upregulates p21WAF1/CIP1 transcription is by preventing the small GTPase RhoA from repressing p21WAF1/CIP1 induction.

* Corresponding author. Mailing address: Drug Discovery Program, H. Lee Moffitt Cancer Center, 12902 Magnolia Dr., Tampa, FL 33612. Phone: (813) 979-6734. Fax: (813) 979-6748. E-mail: sebti{at}moffitt.usf.edu.

Molecular and Cellular Biology, December 1998, p. 6962-6970, Vol. 18, No. 12
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Kazi, A., Carie, A., Blaskovich, M. A., Bucher, C., Thai, V., Moulder, S., Peng, H., Carrico, D., Pusateri, E., Pledger, W. J., Berndt, N., Hamilton, A., Sebti, S. M. (2009). Blockade of Protein Geranylgeranylation Inhibits Cdk2-Dependent p27Kip1 Phosphorylation on Thr187 and Accumulates p27Kip1 in the Nucleus: Implications for Breast Cancer Therapy. Mol. Cell. Biol. 29: 2254-2263 [Abstract] [Full Text]  
  • Watanabe, M., Fiji, H. D. G., Guo, L., Chan, L., Kinderman, S. S., Slamon, D. J., Kwon, O., Tamanoi, F. (2008). Inhibitors of Protein Geranylgeranyltransferase I and Rab Geranylgeranyltransferase Identified from a Library of Allenoate-derived Compounds. J. Biol. Chem. 283: 9571-9579 [Abstract] [Full Text]  
  • Leupold, J. H., Asangani, I., Maurer, G. D., Lengyel, E., Post, S., Allgayer, H. (2007). Src Induces Urokinase Receptor Gene Expression and Invasion/Intravasation via Activator Protein-1/p-c-Jun in Colorectal Cancer. Mol Cancer Res 5: 485-496 [Abstract] [Full Text]  
  • Walker, J. L., Castagnino, P., Chung, B. M., Kazanietz, M. G., Assoian, R. K. (2006). Post-transcriptional Destabilization of p21cip1 by Protein Kinase C in Fibroblasts. J. Biol. Chem. 281: 38127-38132 [Abstract] [Full Text]  
  • Zeng, Y., Zhuang, S., Gloddek, J., Tseng, C.-C., Boss, G. R., Pilz, R. B. (2006). Regulation of cGMP-dependent Protein Kinase Expression by Rho and Kruppel-like Transcription Factor-4. J. Biol. Chem. 281: 16951-16961 [Abstract] [Full Text]  
  • Croft, D. R., Olson, M. F. (2006). The Rho GTPase Effector ROCK Regulates Cyclin A, Cyclin D1, and p27Kip1 Levels by Distinct Mechanisms. Mol. Cell. Biol. 26: 4612-4627 [Abstract] [Full Text]  
  • Wang, D.-A., Sebti, S. M. (2005). Palmitoylated Cysteine 192 Is Required for RhoB Tumor-suppressive and Apoptotic Activities. J. Biol. Chem. 280: 19243-19249 [Abstract] [Full Text]  
  • Patil, S., Bunderson, M., Wilham, J., Black, S. M. (2004). Important role for Rac1 in regulating reactive oxygen species generation and pulmonary arterial smooth muscle cell growth. Am. J. Physiol. Lung Cell. Mol. Physiol. 287: L1314-L1322 [Abstract] [Full Text]  
  • Saika, S., Muragaki, Y., Okada, Y., Miyamoto, T., Ohnishi, Y., Ooshima, A., Kao, W. W.-Y. (2004). Sonic Hedgehog Expression and Role in Healing Corneal Epithelium. IOVS 45: 2577-2585 [Abstract] [Full Text]  
  • Lee, J. Y., Elmer, H. L., Ross, K. R., Kelley, T. J. (2004). Isoprenoid-Mediated Control of SMAD3 Expression in a Cultured Model of Cystic Fibrosis Epithelial Cells. Am. J. Respir. Cell Mol. Bio. 31: 234-240 [Abstract] [Full Text]  
  • Ray, A., Shakya, A., Kumar, D., Ray, B. K. (2004). Overexpression of Serum Amyloid A-Activating Factor 1 Inhibits Cell Proliferation by the Induction of Cyclin-Dependent Protein Kinase Inhibitor p21WAF-1/Cip-1/Sdi-1 Expression. J. Immunol. 172: 5006-5015 [Abstract] [Full Text]  
  • Sun, J., Ohkanda, J., Coppola, D., Yin, H., Kothare, M., Busciglio, B., Hamilton, A. D., Sebti, S. M. (2003). Geranylgeranyltransferase I Inhibitor GGTI-2154 Induces Breast Carcinoma Apoptosis and Tumor Regression in H-Ras Transgenic Mice. Cancer Res. 63: 8922-8929 [Abstract] [Full Text]  
  • Sebti, S. M. (2003). Blocked Pathways: FTIs Shut Down Oncogene Signals. The Oncologist 8: 30-38 [Abstract] [Full Text]  
  • Joyce, P. L., Cox, A. D. (2003). Rac1 and Rac3 Are Targets for Geranylgeranyltransferase I Inhibitor-Mediated Inhibition of Signaling, Transformation, and Membrane Ruffling. Cancer Res. 63: 7959-7967 [Abstract] [Full Text]  
  • Roovers, K., Assoian, R. K. (2003). Effects of Rho Kinase and Actin Stress Fibers on Sustained Extracellular Signal-Regulated Kinase Activity and Activation of G1 Phase Cyclin-Dependent Kinases. Mol. Cell. Biol. 23: 4283-4294 [Abstract] [Full Text]  
  • Doisneau-Sixou, S. F., Cestac, P., Chouini, S., Carroll, J. S., Hamilton, A. D., Sebti, S. M., Poirot, M., Balaguer, P., Faye, J.-C., Sutherland, R. L., Favre, G. (2003). Contrasting Effects of Prenyltransferase Inhibitors on Estrogen-Dependent Cell Cycle Progression and Estrogen Receptor-Mediated Transcriptional Activity in MCF-7 Cells. Endocrinology 144: 989-998 [Abstract] [Full Text]  
  • Chen, J. C., Zhuang, S., Nguyen, T. H., Boss, G. R., Pilz, R. B. (2003). Oncogenic Ras Leads to Rho Activation by Activating the Mitogen-activated Protein Kinase Pathway and Decreasing Rho-GTPase-activating Protein Activity. J. Biol. Chem. 278: 2807-2818 [Abstract] [Full Text]  
  • Sharpe, C. C., Hendry, B. M. (2003). Signaling: Focus on Rho in Renal Disease. J. Am. Soc. Nephrol. 14: 261-264 [Full Text]  
  • Lobell, R. B., Liu, D., Buser, C. A., Davide, J. P., DePuy, E., Hamilton, K., Koblan, K. S., Lee, Y., Mosser, S., Motzel, S. L., Abbruzzese, J. L., Fuchs, C. S., Rowinsky, E. K., Rubin, E. H., Sharma, S., Deutsch, P. J., Mazina, K. E., Morrison, B. W., Wildonger, L., Yao, S.-L., Kohl, N. E. (2002). Preclinical and Clinical Pharmacodynamic Assessment of L-778,123, a Dual Inhibitor of Farnesyl:Protein Transferase and Geranylgeranyl:Protein Transferase Type-I. Molecular Cancer Therapeutics 1: 747-758 [Abstract] [Full Text]  
  • Schwartz, M. A., Assoian, R. K. (2002). Integrins and cell proliferation: regulation of cyclin-dependent kinases via cytoplasmic signaling pathways. J. Cell Sci. 114: 2553-2560 [Abstract] [Full Text]  
  • Adnane, J., Seijo, E., Chen, Z., Bizouarn, F., Leal, M., Sebti, S. M., Munoz-Antonia, T. (2002). RhoB, Not RhoA, Represses the Transcription of the Transforming Growth Factor beta Type II Receptor by a Mechanism Involving Activator Protein 1. J. Biol. Chem. 277: 8500-8507 [Abstract] [Full Text]  
  • Wei, L., Imanaka-Yoshida, K., Wang, L., Zhan, S., Schneider, M. D., DeMayo, F. J., Schwartz, R. J. (2002). Inhibition of Rho family GTPases by Rho GDP dissociation inhibitor disrupts cardiac morphogenesis and inhibits cardiomyocyte proliferation. Development 129: 1705-1714 [Abstract] [Full Text]  
  • Shi, W., Gould, M. N. (2002). Induction of cytostasis in mammary carcinoma cells treated with the anticancer agent perillyl alcohol. Carcinogenesis 23: 131-142 [Abstract] [Full Text]  
  • Lobell, R. B., Omer, C. A., Abrams, M. T., Bhimnathwala, H. G., Brucker, M. J., Buser, C. A., Davide, J. P., deSolms, S. J., Dinsmore, C. J., Ellis-Hutchings, M. S., Kral, A. M., Liu, D., Lumma, W. C., Machotka, S. V., Rands, E., Williams, T. M., Graham, S. L., Hartman, G. D., Oliff, A. I., Heimbrook, D. C., Kohl, N. E. (2001). Evaluation of Farnesyl:Protein Transferase and Geranylgeranyl:Protein Transferase Inhibitor Combinations in Preclinical Models. Cancer Res. 61: 8758-8768 [Abstract] [Full Text]  
  • Han, J.-W., Ahn, S. H., Kim, Y. K., Bae, G.-U., Yoon, J. W., Hong, S., Lee, H. Y., Lee, Y.-W., Lee, H.-W. (2001). Activation of p21WAF1/Cip1 Transcription through Sp1 Sites by Histone Deacetylase Inhibitor Apicidin. INVOLVEMENT OF PROTEIN KINASE C. J. Biol. Chem. 276: 42084-42090 [Abstract] [Full Text]  
  • Adjei, A. A. (2001). Blocking Oncogenic Ras Signaling for Cancer Therapy. JNCI J Natl Cancer Inst 93: 1062-1074 [Abstract] [Full Text]  
  • Reszka, A. A., Halasy-Nagy, J., Rodan, G. A. (2001). Nitrogen-Bisphosphonates Block Retinoblastoma Phosphorylation and Cell Growth by Inhibiting the Cholesterol Biosynthetic Pathway in a Keratinocyte Model for Esophageal Irritation. Mol. Pharmacol. 59: 193-202 [Abstract] [Full Text]  
  • Danen, E. H.J., Sonneveld, P., Sonnenberg, A., Yamada, K. M. (2000). Dual Stimulation of Ras/Mitogen-activated Protein Kinase and RhoA by Cell Adhesion to Fibronectin Supports Growth Factor-stimulated Cell Cycle Progression. JCB 151: 1413-1422 [Abstract] [Full Text]  
  • de Caestecker, M. P., Piek, E., Roberts, A. B. (2000). Role of Transforming Growth Factor-{beta} Signaling in Cancer. JNCI J Natl Cancer Inst 92: 1388-1402 [Abstract] [Full Text]  
  • Park, J.-S., Qiao, L., Gilfor, D., Yang, M. Y., Hylemon, P. B., Benz, C., Darlington, G., Firestone, G., Fisher, P. B., Dent, P. (2000). A Role for Both Ets and C/EBP Transcription Factors and mRNA Stabilization in the MAPK-dependent Increase in p21 Cip-1/WAF1/mda6 Protein Levels in Primary Hepatocytes. Mol. Biol. Cell 11: 2915-2932 [Abstract] [Full Text]  
  • Hu, P. P.-c., Shen, X., Huang, D., Liu, Y., Counter, C., Wang, X.-F. (1999). The MEK Pathway Is Required for Stimulation of p21WAF1/CIP1 by Transforming Growth Factor-beta. J. Biol. Chem. 274: 35381-35387 [Abstract] [Full Text]  
  • Park, J.-S., Carter, S., Reardon, D. B., Schmidt-Ullrich, R., Dent, P., Fisher, P. B. (1999). Roles for Basal and Stimulated p21Cip-1/WAF1/MDA6 Expression and Mitogen-activated Protein Kinase Signaling in Radiation-induced Cell Cycle Checkpoint Control in Carcinoma Cells. Mol. Biol. Cell 10: 4231-4246 [Abstract] [Full Text]  
  • Park, H.-J., Galper, J. B. (1999). 3-Hydroxy-3-methylglutaryl CoA reductase inhibitors up-regulate transforming growth factor-beta signaling in cultured heart cells via inhibition of geranylgeranylation of RhoA GTPase. Proc. Natl. Acad. Sci. USA 96: 11525-11530 [Abstract] [Full Text]  
  • Sowa, Y., Orita, T., Minamikawa-Hiranabe, S., Mizuno, T., Nomura, H., Sakai, T. (1999). Sp3, but not Sp1, Mediates the Transcriptional Activation of the p21/WAF1/Cip1 Gene Promoter by Histone Deacetylase Inhibitor. Cancer Res. 59: 4266-4270 [Abstract] [Full Text]  
  • Sun, J., Qian, Y., Chen, Z., Marfurt, J., Hamilton, A. D., Sebti, S. M. (1999). The Geranylgeranyltransferase I Inhibitor GGTI-298 Induces Hypophosphorylation of Retinoblastoma and Partner Switching of Cyclin-dependent Kinase Inhibitors. A POTENTIAL MECHANISM FOR GGTI-298 ANTITUMOR ACTIVITY. J. Biol. Chem. 274: 6930-6934 [Abstract] [Full Text]  
  • Zhang, W., Geiman, D. E., Shields, J. M., Dang, D. T., Mahatan, C. S., Kaestner, K. H., Biggs, J. R., Kraft, A. S., Yang, V. W. (2000). The Gut-enriched Kruppel-like Factor (Kruppel-like Factor 4) Mediates the Transactivating Effect of p53 on the p21WAF1/Cip1 Promoter. J. Biol. Chem. 275: 18391-18398 [Abstract] [Full Text]  
  • Allal, C., Favre, G., Couderc, B., Salicio, S., Sixou, S., Hamilton, A. D., Sebti, S. M., Lajoie-Mazenc, I., Pradines, A. (2000). RhoA Prenylation Is Required for Promotion of Cell Growth and Transformation and Cytoskeleton Organization but Not for Induction of Serum Response Element Transcription. J. Biol. Chem. 275: 31001-31008 [Abstract] [Full Text]  
  • Huber, H. E., Robinson, R. G., Watkins, A., Nahas, D. D., Abrams, M. T., Buser, C. A., Lobell, R. B., Patrick, D., Anthony, N. J., Dinsmore, C. J., Graham, S. L., Hartman, G. D., Lumma, W. C., Williams, T. M., Heimbrook, D. C. (2001). Anions Modulate the Potency of Geranylgeranyl-Protein Transferase I Inhibitors. J. Biol. Chem. 276: 24457-24465 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»