Journal of Cell Science, Vol 113, Issue 12 2221-2231, Copyright © 2000 by Company of Biologists

JOURNAL ARTICLES

Groucho/TLE/R-esp proteins associate with the nuclear matrix and repress RUNX (CBF(alpha)/AML/PEBP2(alpha)) dependent activation of tissue-specific gene transcription

A Javed, B Guo, S Hiebert, JY Choi, J Green, SC Zhao, MA Osborne, S Stifani, JL Stein, JB Lian, AJ van Wijnen and GS Stein
Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA, USA.

The Runt related transcription factors RUNX (AML/CBF(alpha)/PEBP2(alpha)) are key regulators of hematopoiesis and osteogenesis. Using co-transfection experiments with four natural promoters, including those of the osteocalcin (OC), multi drug resistance (MDR), Rous Sarcoma Virus long terminal repeat (LTR), and bone sialoprotein (BSP) genes, we show that each of these promoters responds differently to the forced expression of RUNX proteins. However, the three RUNX subtypes (i.e. AML1, AML2, and AML3) regulate each promoter in a similar manner. Although the OC promoter is activated in a C terminus dependent manner, the MDR, LTR and BSP promoters are repressed by three distinct mechanisms, either independent of or involving the AML C terminus, or requiring only the conserved C-terminal pentapeptide VWRPY. Using yeast two hybrid assays we find that the C terminus of AML1 interacts with a Groucho/TLE/R-esp repressor protein. Co-expression assays reveal that TLE proteins repress AML dependent activation of OC gene transcription. Western and northern blot analyses suggest that TLE expression is regulated reciprocally with the levels of OC gene expression during osteoblast differentiation. Digital immunofluorescence microscopy results show that TLE1 and TLE2 are both associated with the nuclear matrix, and that a significant subset of each colocalizes with AML transcription factors. This co-localization of TLE and AML proteins is lost upon removing the C terminus of AML family members. Our findings indicate that suppression of AML-dependent gene activation by TLE proteins involves functional interactions with the C terminus of AML at the nuclear matrix in situ. Our data are consistent with the concept that the C termini of AML proteins support activation or repression of cell-type specific genes depending on the regulatory organization of the target promoter and subnuclear localization.

This article has been cited by other articles:

				L. J. Marshall, A. C. Moore, M. Ohki, I. Kitabayashi, D. Patterson, and D. A. Ornelles RUNX1 Permits E4orf6-Directed Nuclear Localization of the Adenovirus E1B-55K Protein and Associates with Centers of Viral DNA and RNA Synthesis J. Virol., July 1, 2008; 82(13): 6395 - 6408. [Abstract] [Full Text] [PDF]

				A. Javed, J.-S. Bae, F. Afzal, S. Gutierrez, J. Pratap, S. K. Zaidi, Y. Lou, A. J. van Wijnen, J. L. Stein, G. S. Stein, et al. Structural Coupling of Smad and Runx2 for Execution of the BMP2 Osteogenic Signal J. Biol. Chem., March 28, 2008; 283(13): 8412 - 8422. [Abstract] [Full Text] [PDF]

				V. Lamour, C. Detry, C. Sanchez, Y. Henrotin, V. Castronovo, and A. Bellahcene Runx2- and Histone Deacetylase 3-mediated Repression Is Relieved in Differentiating Human Osteoblast Cells to Allow High Bone Sialoprotein Expression J. Biol. Chem., December 14, 2007; 282(50): 36240 - 36249. [Abstract] [Full Text] [PDF]

				S. K. Zaidi, S. Pande, J. Pratap, T. Gaur, S. Grigoriu, S. A. Ali, J. L. Stein, J. B. Lian, A. J. van Wijnen, and G. S. Stein Runx2 deficiency and defective subnuclear targeting bypass senescence to promote immortalization and tumorigenic potential PNAS, December 11, 2007; 104(50): 19861 - 19866. [Abstract] [Full Text] [PDF]

				T. Egawa, R. E. Tillman, Y. Naoe, I. Taniuchi, and D. R. Littman The role of the Runx transcription factors in thymocyte differentiation and in homeostasis of naive T cells J. Exp. Med., August 6, 2007; 204(8): 1945 - 1957. [Abstract] [Full Text] [PDF]

				M. Q. Hassan, R. Tare, S. H. Lee, M. Mandeville, B. Weiner, M. Montecino, A. J. van Wijnen, J. L. Stein, G. S. Stein, and J. B. Lian HOXA10 Controls Osteoblastogenesis by Directly Activating Bone Regulatory and Phenotypic Genes Mol. Cell. Biol., May 1, 2007; 27(9): 3337 - 3352. [Abstract] [Full Text] [PDF]

				Y. Babaie, R. Herwig, B. Greber, T. C. Brink, W. Wruck, D. Groth, H. Lehrach, T. Burdon, and J. Adjaye Analysis of Oct4-Dependent Transcriptional Networks Regulating Self-Renewal and Pluripotency in Human Embryonic Stem Cells Stem Cells, February 1, 2007; 25(2): 500 - 510. [Abstract] [Full Text] [PDF]

				T. Heimbucher, C. Murko, B. Bajoghli, N. Aghaallaei, A. Huber, R. Stebegg, D. Eberhard, M. Fink, A. Simeone, and T. Czerny Gbx2 and Otx2 Interact with the WD40 Domain of Groucho/Tle Corepressors Mol. Cell. Biol., January 1, 2007; 27(1): 340 - 351. [Abstract] [Full Text] [PDF]

				C. S. Shin, M. J. Jeon, J.-Y. Yang, S.-J. Her, D. Kim, S. W. Kim, and S. Y. Kim CCAAT/enhancer-binding protein {delta} activates the Runx2-mediated transcription of mouse osteocalcin II promoter. J. Mol. Endocrinol., June 1, 2006; 36(3): 531 - 546. [Abstract] [Full Text] [PDF]

				K. S. Moorefield, H. Yin, T. D. Nichols, C. Cathcart, S. O. Simmons, and J. M. Horowitz Sp2 Localizes to Subnuclear Foci Associated with the Nuclear Matrix Mol. Biol. Cell, April 1, 2006; 17(4): 1711 - 1722. [Abstract] [Full Text] [PDF]

				Q. Shen and S. Christakos The Vitamin D Receptor, Runx2, and the Notch Signaling Pathway Cooperate in the Transcriptional Regulation of Osteopontin J. Biol. Chem., December 9, 2005; 280(49): 40589 - 40598. [Abstract] [Full Text] [PDF]

				L. F. Peterson, A. Boyapati, V. Ranganathan, A. Iwama, D. G. Tenen, S. Tsai, and D.-E. Zhang The Hematopoietic Transcription Factor AML1 (RUNX1) Is Negatively Regulated by the Cell Cycle Protein Cyclin D3 Mol. Cell. Biol., December 1, 2005; 25(23): 10205 - 10219. [Abstract] [Full Text] [PDF]

				C. Sakakura, K. Hasegawa, K. Miyagawa, S. Nakashima, T. Yoshikawa, S. Kin, Y. Nakase, S. Yazumi, H. Yamagishi, T. Okanoue, et al. Possible Involvement of RUNX3 Silencing in the Peritoneal Metastases of Gastric Cancers Clin. Cancer Res., September 15, 2005; 11(18): 6479 - 6488. [Abstract] [Full Text] [PDF]

				G. Xiao, D. Jiang, C. Ge, Z. Zhao, Y. Lai, H. Boules, M. Phimphilai, X. Yang, G. Karsenty, and R. T. Franceschi Cooperative Interactions between Activating Transcription Factor 4 and Runx2/Cbfa1 Stimulate Osteoblast-specific Osteocalcin Gene Expression J. Biol. Chem., September 2, 2005; 280(35): 30689 - 30696. [Abstract] [Full Text] [PDF]

				Y. Fukushima-Nakase, Y. Naoe, I. Taniuchi, H. Hosoi, T. Sugimoto, and T. Okuda Shared and distinct roles mediated through C-terminal subdomains of acute myeloid leukemia/Runt-related transcription factor molecules in murine development Blood, June 1, 2005; 105(11): 4298 - 4307. [Abstract] [Full Text] [PDF]

				M. Galindo, J. Pratap, D. W. Young, H. Hovhannisyan, H.-J. Im, J.-Y. Choi, J. B. Lian, J. L. Stein, G. S. Stein, and A. J. van Wijnen The Bone-specific Expression of Runx2 Oscillates during the Cell Cycle to Support a G1-related Antiproliferative Function in Osteoblasts J. Biol. Chem., May 27, 2005; 280(21): 20274 - 20285. [Abstract] [Full Text] [PDF]

				D. Vradii, S. K. Zaidi, J. B. Lian, A. J. van Wijnen, J. L. Stein, and G. S. Stein Point mutation in AML1 disrupts subnuclear targeting, prevents myeloid differentiation, and effects a transformation-like phenotype PNAS, May 17, 2005; 102(20): 7174 - 7179. [Abstract] [Full Text] [PDF]

				H.-I. Trompeter, N. Gomez-Lozano, S. Santourlidis, B. Eisermann, P. Wernet, C. Vilches, and M. Uhrberg Three Structurally and Functionally Divergent Kinds of Promoters Regulate Expression of Clonally Distributed Killer Cell Ig-Like Receptors (KIR), of KIR2DL4, and of KIR3DL3 J. Immunol., April 1, 2005; 174(7): 4135 - 4143. [Abstract] [Full Text] [PDF]

				A. Javed, G. L. Barnes, J. Pratap, T. Antkowiak, L. C. Gerstenfeld, A. J. van Wijnen, J. L. Stein, J. B. Lian, and G. S. Stein Impaired intranuclear trafficking of Runx2 (AML3/CBFA1) transcription factors in breast cancer cells inhibits osteolysis in vivo PNAS, February 1, 2005; 102(5): 1454 - 1459. [Abstract] [Full Text] [PDF]

				S. K. Zaidi, D. W. Young, J.-Y. Choi, J. Pratap, A. Javed, M. Montecino, J. L. Stein, J. B. Lian, A. J. van Wijnen, and G. S. Stein Intranuclear Trafficking: Organization and Assembly of Regulatory Machinery for Combinatorial Biological Control J. Biol. Chem., October 15, 2004; 279(42): 43363 - 43366. [Abstract] [Full Text] [PDF]

				H. N. Nuthall, K. Joachim, and S. Stifani Phosphorylation of Serine 239 of Groucho/TLE1 by Protein Kinase CK2 Is Important for Inhibition of Neuronal Differentiation Mol. Cell. Biol., October 1, 2004; 24(19): 8395 - 8407. [Abstract] [Full Text] [PDF]

				T. M. Schroeder, R. A. Kahler, X. Li, and J. J. Westendorf Histone Deacetylase 3 Interacts with Runx2 to Repress the Osteocalcin Promoter and Regulate Osteoblast Differentiation J. Biol. Chem., October 1, 2004; 279(40): 41998 - 42007. [Abstract] [Full Text] [PDF]

				D. W. Young, S. K. Zaidi, P. S. Furcinitti, A. Javed, A. J. van Wijnen, J. L. Stein, J. B. Lian, and G. S. Stein Quantitative signature for architectural organization of regulatory factors using intranuclear informatics J. Cell Sci., October 1, 2004; 117(21): 4889 - 4896. [Abstract] [Full Text] [PDF]

				G. L. Barnes, K. E. Hebert, M. Kamal, A. Javed, T. A. Einhorn, J. B. Lian, G. S. Stein, and L. C. Gerstenfeld Fidelity of Runx2 Activity in Breast Cancer Cells Is Required for the Generation of Metastases-Associated Osteolytic Disease Cancer Res., July 1, 2004; 64(13): 4506 - 4513. [Abstract] [Full Text] [PDF]

				T. Yoshizawa, F. Takizawa, F. Iizawa, O. Ishibashi, H. Kawashima, A. Matsuda, N. Endo, and H. Kawashima Homeobox Protein Msx2 Acts as a Molecular Defense Mechanism for Preventing Ossification in Ligament Fibroblasts Mol. Cell. Biol., April 15, 2004; 24(8): 3460 - 3472. [Abstract] [Full Text] [PDF]

				M. Nishimura, Y. Fukushima-Nakase, Y. Fujita, M. Nakao, S. Toda, N. Kitamura, T. Abe, and T. Okuda VWRPY motif-dependent and -independent roles of AML1/Runx1 transcription factor in murine hematopoietic development Blood, January 15, 2004; 103(2): 562 - 570. [Abstract] [Full Text] [PDF]

				F. LOMBARDO, D. KOMATSU, and M. HADJIARGYROU Molecular cloning and characterization of Mustang, a novel nuclear protein expressed during skeletal development and regeneration FASEB J, January 1, 2004; 18(1): 52 - 61. [Abstract] [Full Text] [PDF]

				S. K. Zaidi, D. W. Young, S. M. Pockwinse, A. Javed, J. B. Lian, J. L. Stein, A. J. van Wijnen, and G. S. Stein Mitotic partitioning and selective reorganization of tissue-specific transcription factors in progeny cells PNAS, December 9, 2003; 100(25): 14852 - 14857. [Abstract] [Full Text] [PDF]

				M. Ehlers, K. Laule-Kilian, M. Petter, C. J. Aldrian, B. Grueter, A. Wurch, N. Yoshida, T. Watanabe, M. Satake, and V. Steimle Morpholino Antisense Oligonucleotide-Mediated Gene Knockdown During Thymocyte Development Reveals Role for Runx3 Transcription Factor in CD4 Silencing During Development of CD4-/CD8+ Thymocytes J. Immunol., October 1, 2003; 171(7): 3594 - 3604. [Abstract] [Full Text] [PDF]

				M. J. Jeon, J. A. Kim, S. H. Kwon, S. W. Kim, K. S. Park, S.-W. Park, S. Y. Kim, and C. S. Shin Activation of Peroxisome Proliferator-activated Receptor-{gamma} Inhibits the Runx2-mediated Transcription of Osteocalcin in Osteoblasts J. Biol. Chem., June 20, 2003; 278(26): 23270 - 23277. [Abstract] [Full Text] [PDF]

				C. A. P. Bristow and P. Shore Transcriptional regulation of the human MIP-1{alpha} promoter by RUNX1 and MOZ Nucleic Acids Res., June 1, 2003; 31(11): 2735 - 2744. [Abstract] [Full Text] [PDF]

				J. Shen, H. Hovhannisyan, J. B. Lian, M. A. Montecino, G. S. Stein, J. L. Stein, and A. J. van Wijnen Transcriptional Induction of the Osteocalcin Gene During Osteoblast Differentiation Involves Acetylation of Histones H3 and H4 Mol. Endocrinol., April 1, 2003; 17(4): 743 - 756. [Abstract] [Full Text] [PDF]

				H. N. Nuthall, K. Joachim, A. Palaparti, and S. Stifani A Role for Cell Cycle-regulated Phosphorylation in Groucho-mediated Transcriptional Repression J. Biol. Chem., December 20, 2002; 277(52): 51049 - 51057. [Abstract] [Full Text] [PDF]

				M. Lepourcelet and R. A. Shivdasani Characterization of a Novel Mammalian Groucho Isoform and Its Role in Transcriptional Regulation J. Biol. Chem., November 27, 2002; 277(49): 47732 - 47740. [Abstract] [Full Text] [PDF]

				J. J. Westendorf, S. K. Zaidi, J. E. Cascino, R. Kahler, A. J. van Wijnen, J. B. Lian, M. Yoshida, G. S. Stein, and X. Li Runx2 (Cbfa1, AML-3) Interacts with Histone Deacetylase 6 and Represses the p21CIP1/WAF1 Promoter Mol. Cell. Biol., November 15, 2002; 22(22): 7982 - 7992. [Abstract] [Full Text] [PDF]

				S. K. Zaidi, A. J. Sullivan, A. J. van Wijnen, J. L. Stein, G. S. Stein, and J. B. Lian Integration of Runx and Smad regulatory signals at transcriptionally active subnuclear sites PNAS, June 11, 2002; 99(12): 8048 - 8053. [Abstract] [Full Text] [PDF]

				H. N. Nuthall, J. Husain, K. W. McLarren, and S. Stifani Role for Hes1-Induced Phosphorylation in Groucho-Mediated Transcriptional Repression Mol. Cell. Biol., January 15, 2002; 22(2): 389 - 399. [Abstract] [Full Text] [PDF]

				K. S. Harrington, A. Javed, H. Drissi, S. McNeil, J. B. Lian, J. L. Stein, A. J. van Wijnen, Y.-L. Wang, and G. S. Stein Transcription factors RUNX1/AML1 and RUNX2/Cbfa1 dynamically associate with stationary subnuclear domains J. Cell Sci., January 11, 2002; 115(21): 4167 - 4176. [Abstract] [Full Text] [PDF]

				R. C. D'Alonzo, N. Selvamurugan, G. Karsenty, and N. C. Partridge Physical Interaction of the Activator Protein-1 Factors c-Fos and c-Jun with Cbfa1 for Collagenase-3 Promoter Activation J. Biol. Chem., January 4, 2002; 277(1): 816 - 822. [Abstract] [Full Text]

				W. Liu, S. Toyosawa, T. Furuichi, N. Kanatani, C. Yoshida, Y. Liu, M. Himeno, S. Narai, A. Yamaguchi, and T. Komori Overexpression of Cbfa1 in osteoblasts inhibits osteoblast maturation and causes osteopenia with multiple fractures J. Cell Biol., October 1, 2001; 155(1): 157 - 166. [Abstract] [Full Text] [PDF]

				C. Banerjee, A. Javed, J.-Y. Choi, J. Green, V. Rosen, A. J. van Wijnen, J. L. Stein, J. B. Lian, and G. S. Stein Differential Regulation of the Two Principal Runx2/Cbfa1 N-Terminal Isoforms in Response to Bone Morphogenetic Protein-2 during Development of the Osteoblast Phenotype Endocrinology, September 1, 2001; 142(9): 4026 - 4039. [Abstract] [Full Text] [PDF]

				J.-Y. Choi, J. Pratap, A. Javed, S. K. Zaidi, L. Xing, E. Balint, S. Dalamangas, B. Boyce, A. J. van Wijnen, J. B. Lian, et al. Subnuclear targeting of Runx/Cbfa/AML factors is essential for tissue-specific differentiation during embryonic development PNAS, June 28, 2001; (2001) 151236498. [Abstract] [Full Text] [PDF]

				A. Javed, G. L. Barnes, B. O. Jasanya, J. L. Stein, L. Gerstenfeld, J. B. Lian, and G. S. Stein runt Homology Domain Transcription Factors (Runx, Cbfa, and AML) Mediate Repression of the Bone Sialoprotein Promoter: Evidence for Promoter Context-Dependent Activity of Cbfa Proteins Mol. Cell. Biol., April 15, 2001; 21(8): 2891 - 2905. [Abstract] [Full Text]

				S. K. Zaidi, A. Javed, J.-Y. Choi, A. J. van Wijnen, J. L. Stein, J. B. Lian, and G. S. Stein A specific targeting signal directs Runx2/Cbfa1 to subnuclear domains and contributes to transactivation of the osteocalcin gene J. Cell Sci., January 9, 2001; 114(17): 3093 - 3102. [Abstract] [Full Text] [PDF]

				K. W. McLarren, F. M. Theriault, and S. Stifani Association with the Nuclear Matrix and Interaction with Groucho and RUNX Proteins Regulate the Transcription Repression Activity of the Basic Helix Loop Helix Factor Hes1 J. Biol. Chem., January 5, 2001; 276(2): 1578 - 1584. [Abstract] [Full Text] [PDF]

				J. Hess, D. Porte, C. Munz, and P. Angel AP-1 and Cbfa/Runt Physically Interact and Regulate Parathyroid Hormone-dependent MMP13 Expression in Osteoblasts through a New Osteoblast-specific Element 2/AP-1 Composite Element J. Biol. Chem., June 1, 2001; 276(23): 20029 - 20038. [Abstract] [Full Text] [PDF]

				S. Gutierrez, A. Javed, D. K. Tennant, M. van Rees, M. Montecino, G. S. Stein, J. L. Stein, and J. B. Lian CCAAT/Enhancer-binding Proteins (C/EBP) beta and delta Activate Osteocalcin Gene Transcription and Synergize with Runx2 at the C/EBP Element to Regulate Bone-specific Expression J. Biol. Chem., January 4, 2002; 277(2): 1316 - 1323. [Abstract] [Full Text] [PDF]

				J.-Y. Choi, J. Pratap, A. Javed, S. K. Zaidi, L. Xing, E. Balint, S. Dalamangas, B. Boyce, A. J. van Wijnen, J. B. Lian, et al. Subnuclear targeting of Runx/Cbfa/AML factors is essential for tissue-specific differentiation during embryonic development PNAS, July 17, 2001; 98(15): 8650 - 8655. [Abstract] [Full Text] [PDF]