HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kurokawa, M. Articles by Hirai, H. Search for Related Content PubMed PubMed Citation Articles by Kurokawa, M. Articles by Hirai, H. Social Bookmarking                      What's this?

Volume 271, Number 28, Issue of July 12, 1996 pp. 16870-16876
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

---

A Conserved Cysteine Residue in the runt Homology Domain of AML1 Is Required for the DNA Binding Ability and the Transforming Activity on Fibroblasts

(Received for publication, January 25, 1996)

From the Third Department of Internal Medicine, Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113, Japan

The AML1 gene encodes DNA-binding proteins that contain the runt homology domain and is found at the breakpoints of t(8;21), t(3;21), and t(12;21) translocations associated with myelogenous leukemias. AML1 heterodimerizes with PEBP2/CBF, resulting in the enhanced affinity with DNA. The runt homology domain is responsible for binding with DNA and heterodimerizing with PEBP2/CBF. AML1 is suggested to perform a pivotal role in myeloid cell differentiation, whereas it can cause neoplastic transformation when overexpressed in fibroblasts. In this study, we demonstrated that the reducing reagent, dithiothreitol (DTT), markedly enhances the DNA binding of AML1 expressed in COS7 cells. Oxidation by diamide or modification by N-ethylmaleimide of the free sulfhydryl residues inhibited the interaction of AML1 with DNA. The diamide effect was reversible with excess of DTT, whereas DTT could not restore the DNA binding of AML1 treated with N-ethylmaleimide. Site-directed mutagenesis of the amino acid residue 72, a highly conserved cysteine in the runt homology domain of AML1, to serine almost completely abolished DNA binding without altering the interaction with PEBP2/CBF. This substitution also impaired transactivation through the consensus DNA sequence and transformation of fibroblasts induced by AML1b. These data indicate an essential role of the conserved cysteine residue in DNA binding of AML1, and it is possible that the redox state of AML1 could contribute to the regulation of its function.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Jo and T. E. Curry Jr. Luteinizing Hormone-Induced RUNX1 Regulates the Expression of Genes in Granulosa Cells of Rat Periovulatory Follicles Mol. Endocrinol., September 1, 2006; 20(9): 2156 - 2172. [Abstract] [Full Text] [PDF]

				M. Jo, M. C. Gieske, C. E. Payne, S. E. Wheeler-Price, J. B. Gieske, I. V. Ignatius, T. E. Curry Jr., and C. Ko Development and Application of a Rat Ovarian Gene Expression Database Endocrinology, November 1, 2004; 145(11): 5384 - 5396. [Abstract] [Full Text] [PDF]

				F.-Q. Li, R. E. Person, K.-I. Takemaru, K. Williams, K. Meade-White, A. H. Ozsahin, T. Gungor, R. T. Moon, and M. Horwitz Lymphoid Enhancer Factor-1 Links Two Hereditary Leukemia Syndromes through Core-binding Factor {alpha} Regulation of ELA2 J. Biol. Chem., January 23, 2004; 279(4): 2873 - 2884. [Abstract] [Full Text] [PDF]

				H. Sakoda, Y. Gotoh, H. Katagiri, M. Kurokawa, H. Ono, Y. Onishi, M. Anai, T. Ogihara, M. Fujishiro, Y. Fukushima, et al. Differing Roles of Akt and Serum- and Glucocorticoid-regulated Kinase in Glucose Metabolism, DNA Synthesis, and Oncogenic Activity J. Biol. Chem., July 3, 2003; 278(28): 25802 - 25807. [Abstract] [Full Text] [PDF]

				P. Ducy, M. Starbuck, M. Priemel, J. Shen, G. Pinero, V. Geoffroy, M. Amling, and G. Karsenty A Cbfa1-dependent genetic pathway controls bone formation beyond embryonic development Genes & Dev., April 15, 1999; 13(8): 1025 - 1036. [Abstract] [Full Text]

				M. Osato, N. Asou, E. Abdalla, K. Hoshino, H. Yamasaki, T. Okubo, H. Suzushima, K. Takatsuki, T. Kanno, K. Shigesada, et al. Biallelic and Heterozygous Point Mutations in the Runt Domain of the AML1/PEBP2alpha B Gene Associated With Myeloblastic Leukemias Blood, March 15, 1999; 93(6): 1817 - 1824. [Abstract] [Full Text] [PDF]

				S. Kramer, T. Jinks, P Schedl, and J. Gergen Direct activation of Sex-lethal transcription by the Drosophila runt protein Development, January 1, 1999; 126(1): 191 - 200. [Abstract] [PDF]

				K. Thirunavukkarasu, M. Mahajan, K. W. McLarren, S. Stifani, and G. Karsenty Two Domains Unique to Osteoblast-Specific Transcription Factor Osf2/Cbfa1 Contribute to Its Transactivation Function and Its Inability To Heterodimerize with Cbfbeta Mol. Cell. Biol., July 1, 1998; 18(7): 4197 - 4208. [Abstract] [Full Text]

				N. Selvamurugan, W.-Y. Chou, A. T. Pearman, M. R. Pulumati, and N. C. Partridge Parathyroid Hormone Regulates the Rat Collagenase-3 Promoter in Osteoblastic Cells through the Cooperative Interaction of the Activator Protein-1 Site and the runt Domain Binding Sequence J. Biol. Chem., April 24, 1998; 273(17): 10647 - 10657. [Abstract] [Full Text] [PDF]

				K. Tanaka, T. Tanaka, M. Kurokawa, Y. Imai, S. Ogawa, K. Mitani, Y. Yazaki, and H. Hirai The AML1/ETO(MTG8) and AML1/Evi-1 Leukemia-Associated Chimeric Oncoproteins Accumulate PEBP2beta (CBFbeta ) in the Nucleus More Efficiently Than Wild-Type AML1 Blood, March 1, 1998; 91(5): 1688 - 1699. [Abstract] [Full Text] [PDF]

				C. Zeng, A. J. van Wijnen, J. L. Stein, S. Meyers, W. Sun, L. Shopland, J. B. Lawrence, S. Penman, J. B. Lian, G. S. Stein, et al. Identification of a nuclear matrix targeting signal in the leukemia and bone-related AML/CBF-alpha transcription factors PNAS, June 24, 1997; 94(13): 6746 - 6751. [Abstract] [Full Text] [PDF]

				Y. Akamatsu, T. Ohno, K. Hirota, H. Kagoshima, J. Yodoi, and K. Shigesada Redox Regulation of the DNA Binding Activity in Transcription Factor PEBP2. THE ROLES OF TWO CONSERVED CYSTEINE RESIDUES J. Biol. Chem., June 6, 1997; 272(23): 14497 - 14500. [Abstract] [Full Text] [PDF]

				K. Thirunavukkarasu, D. L. Halladay, R. R. Miles, X. Yang, R. J. S. Galvin, S. Chandrasekhar, T. J. Martin, and J. E. Onyia The Osteoblast-specific Transcription Factor Cbfa1 Contributes to the Expression of Osteoprotegerin, a Potent Inhibitor of Osteoclast Differentiation and Function J. Biol. Chem., August 11, 2000; 275(33): 25163 - 25172. [Abstract] [Full Text] [PDF]

				B. Kern, J. Shen, M. Starbuck, and G. Karsenty Cbfa1 Contributes to the Osteoblast-specific Expression of type I collagen Genes J. Biol. Chem., March 2, 2001; 276(10): 7101 - 7107. [Abstract] [Full Text] [PDF]