Knowledge of the transcriptional circuitry responsible for pluripotentiality and self-renewal in embryonic stem cells is tantamount to understanding early mammalian development and a prerequisite to determining their therapeutic potential. Various techniques have employed genomics to identify transcripts that were abundant in stem cells, in an attempt to define the molecular basis of ‘stemness’. In this study, we have extended traditional genomic analyses to identify cis-elements that might be implicated in the control of embryonic stem cell-restricted gene promoters. The strategy relied on the generation of a problem-specific list from serial analysis of gene expression profiles and subsequent promoter analyses to identify frameworks of multiple cis-elements conserved in space and orientation among genes from the problem-specific list. Subsequent experimental data suggest that 2 novel transcription factors, B-Myb and Maz, predicted from these models, are implicated either in the maintenance of the undifferentiated stem cell state or in early steps of differentiation.

1.
Anisimov, S.V., K.V. Tarasov, D. Tweedie, M.D. Stern, A.M. Wobus, K.R. Boheler (2002) SAGE identification of gene transcripts with profiles unique to pluripotent mouse R1 embryonic stem cells. Genomics 79: 169–176.
2.
Bhattacharya, B., T. Miura, R. Brandenberger, J. Mejido, Y. Luo, A.X. Yang, B.H. Joshi, I. Ginis, R.S. Thies, M. Amit, I. Lyons, B.G. Condie, J. Itskovitz-Eldor, M.S. Rao, R.K. Puri (2004) Gene expression in human embryonic stem cell lines: unique molecular signature. Blood 103: 2956–2964.
3.
Bossone, S.A., C. Asselin, A.J. Patel, Marcu K.B. (1992) MAZ, a zinc finger protein, binds to c-MYC and C2 gene sequences regulating transcriptional initiation and termination. Proc Natl Acad Sci USA 89: 7452–7456.
4.
Boyer, L.A., T.I. Lee, M.F. Cole, S.E. Johnstone, S.S. Levine, J.P. Zucker, M.G. Guenther, R.M. Kumar, H.L. Murray, R.G. Jenner, D.K. Gifford, D.A. Melton, R. Jaenisch, R.A. Young (2005) Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 122: 947–956.
5.
Cartharius, K., K. Frech, K. Grote, B. Klocke, M. Haltmeier, A. Klingenhoff, M. Frisch, M. Bayerlein, T. Werner (2005) MatInspector and beyond: promoter analysis based on transcription factor binding sites. Bioinformatics 21: 2933–2942.
6.
Cartwright, P., C. McLean, A. Sheppard, D. Rivett, K. Jones, S. Dalton (2005) LIF/STAT3 controls ES cell self-renewal and pluripotency by a Myc-dependent mechanism. Development 132: 885–896.
7.
Doetschman, T.C., H. Eistetter, M. Katz, W. Schmidt, R. Kemler (1985) The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morphol 87: 27–45.
8.
Dohr, S., A. Klingenhoff, H. Maier, M. Hrabe de Angelis, T. Werner, R. Schneider (2005) Linking disease-associated genes to regulatory networks via promoter organization. Nucleic Acids Res 33: 864–872.
9.
Gan, Q., T. Yoshida, O.G. McDonald, G.K. Owens (2007) Concise review: epigenetic mechanisms contribute to pluripotency and cell lineage determination of embryonic stem cells. Stem Cells 25: 2–9.
10.
Ginis, I., Y. Luo, T. Miura, S. Thies, R. Brandenberger, S. Gerecht-Nir, M. Amit, A. Hoke, M.K. Carpenter, J. Itskovitz-Eldor, M.S. Rao (2004) Differences between human and mouse embryonic stem cells. Dev Biol 269: 360–380.
11.
Iwai, N., K. Kitajima, K. Sakai, T. Kimura, T. Nakano (2001) Alteration of cell adhesion and cell cycle properties of ES cells by an inducible dominant interfering Myb mutant. Oncogene 20: 1425–1434.
12.
Joaquin, M., R.J. Watson (2003) Cell cycle regulation by the B-Myb transcription factor. Cell Mol Life Sci 60: 2389–2401.
13.
Johnson, L.R., T.K. Johnson, M. Desler, T.A. Luster, T. Nowling, R.E. Lewis, A. Rizzino (2002) Effects of B-Myb on gene transcription: phosphorylation-dependent activity and acetylation by p300. J Biol Chem 277: 4088–4097.
14.
Keller, G. (2005) Embryonic stem cell differentiation: emergence of a new era in biology and medicine. Genes Dev 19: 1129–1155.
15.
Komatsu, M., H.O. Li, H. Tsutsui, K. Itakura, M. Matsumura, K.K. Yokoyama (1997) MAZ, a Myc-associated zinc finger protein, is essential for the ME1a1-mediated expression of the c-myc gene during neuroectodermal differentiation of P19 cells. Oncogene 15: 1123–1131.
16.
Lee, J.H., S.R. Hart, D.G. Skalnik (2004) Histone deacetylase activity is required for embryonic stem cell differentiation. Genesis 38: 32–38.
17.
Nagy, A., J. Rossant, R. Nagy, W. Abramow-Newerly, J.C. Roder (1993) Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc Natl Acad Sci USA 90: 8424–8428.
18.
Nakagoshi, H., C. Kanei-Ishii, T. Sawazaki, G. Mizuguchi, S. Ishii (1992) Transcriptional activation of the c-myc gene by the c-myb and B-myb gene products. Oncogene 7: 1233–1240.
19.
Rathjen, J., J.A. Lake, M.D. Bettess, J.M. Washington, G. Chapman, P.D. Rathjen (1999) Formation of a primitive ectoderm like cell population, EPL cells, from ES cells in response to biologically derived factors. J Cell Sci 112: 601–612.
20.
Richards, M., S.P. Tan, J.H. Tan, W.K. Chan, A. Bongso (2004) The transcriptome profile of human embryonic stem cells as defined by SAGE. Stem Cells 22: 51–64.
21.
Sala, A., R. Watson (1999) B-Myb protein in cellular proliferation, transcription control, and cancer: latest developments. J Cell Physiol 179: 245–250.
22.
Sala, A. (2005) B-MYB, a transcription factor implicated in regulating cell cycle, apoptosis and cancer. Eur J Cancer 41: 2479–2484.
23.
Sandberg, M.L., S.E. Sutton, M.T. Pletcher, T. Wiltshire, L.M. Tarantino, J.B. Hogenesch, M.P. Cooke (2005) c-Myb and p300 regulate hematopoietic stem cell proliferation and differentiation. Dev Cell 8: 153–166.
24.
Sato, N., I.M. Sanjuan, M. Heke, M. Uchida, F. Naef, A.H. Brivanlou (2003) Molecular signature of human embryonic stem cells and its comparison with the mouse. Dev Biol 260: 404–413.
25.
Seifert, M., M. Scherf, A. Epple, T. Werner (2005) Multievidence microarray mining. Trends Genet 21: 553–558.
26.
Song, J., H. Ugai, H. Nakata-Tsutsui, S. Kishikawa, E. Suzuki, T. Murata, K.K. Yokoyama (2003) Transcriptional regulation by zinc-finger proteins Sp1 and MAZ involves interactions with the same cis-elements. Int J Mol Med 11: 547–553.
27.
Sperger, J.M., X. Chen, J.S. Draper, J.E. Antosiewicz, C.H. Chon, S.B. Jones, J.D. Brooks, P.W. Andrews, P.O. Brown, J.A. Thomson (2003) Gene expression patterns in human embryonic stem cells and human pluripotent germ cell tumors. Proc Natl Acad Sci USA 100: 13350–13355.
28.
Stewart, C.L., I. Gadi, H. Bhatt (1994) Stem cells from primordial germ cells can reenter the germ line. Dev Biol 161: 626–628.
29.
Tanaka, Y., N.P. Patestos, T. Maekawa, S. Ishii (1999) B-myb is required for inner cell mass formation at an early stage of development. J Biol Chem 274: 28067–28070.
30.
Tarasova, Y.S., D.R. Riordon, K.V. Tarasov, K.R. Boheler (2006) In vitro differentiation of mouse ES cells to muscle cells; in Notarianni, E., M.J. Evans (eds): Embryonic Stem Cells: A Practical Approach. New York, Oxford University Press, pp 130–168.
31.
Tashiro, S., Y. Takemoto, H. Handa, S. Ishii (1995) Cell type-specific trans-activation by the B-myb gene product: requirement of the putative cofactor binding to the C-terminal conserved domain. Oncogene 10: 1699–1707.
32.
Tashiro, S., T. Sumi, N. Uozumi, T. Shimizu, T. Nakamura (2004) B-Myb-dependent regulation of c-Myc expression by cytosolic phospholipase A2. J Biol Chem 279: 17715–17722.
33.
Thomson, J.A., J. Itskovitz-Eldor, S.S. Shapiro, M.A. Waknitz, J.J. Swiergiel, V.S. Marshall, J.M. Jones (1998) Embryonic stem cell lines derived from human blastocysts. Science 282: 1145–1147.
34.
Toscani, A., R.V. Mettus, R. Coupland, H. Simpkins, J. Litvin, J. Orth, K.S. Hatton, E.P. Reddy (1997) Arrest of spermatogenesis and defective breast development in mice lacking A-myb. Nature 386: 713–717.
35.
Velculescu, V.E., L. Zhang, B. Vogelstein, K.W. Kinzler (1995) Serial analysis of gene expression. Science 270: 484–487.
36.
Wang, J., S. Rao, J. Chu, X. Shen, D.N. Levasseur, T.W. Theunissen, S.H. Orkin (2006) A protein interaction network for pluripotency of embryonic stem cells. Nature 444: 364–368.
37.
Werner, T. (2000) Identification and functional modelling of DNA sequence elements of transcription. Brief Bioinform 1: 372–380.
38.
Werner, T., S. Fessele, H. Maier, P.J. Nelson (2003) Computer modeling of promoter organization as a tool to study transcriptional coregulation. FASEB J 17: 1228–1237.
39.
Wobus, A.M., K. Guan, H.-T. Yang, K.R. Boheler (2002) Embryonic stem cells as a model to study cardiac, skeletal muscle and vascular smooth muscle cell differentiation. Methods Mol Biol 185: 127–156.
40.
Wobus, A.M., K.R. Boheler (2005) Embryonic stem cells: prospects for developmental biology and cell therapy. Physiol Rev 85: 635–678.
41.
Yamane, T., S.J. Dylla, M. Muijtjens, I.L. Weissman (2005) Enforced Bcl-2 expression overrides serum and feeder cell requirements for mouse embryonic stem cell self-renewal. Proc Natl Acad Sci USA 102: 3312–3317.
42.
Ying, Q.L., J. Nichols, I. Chambers, A. Smith (2003) BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell 115: 281–292.
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