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J. Biol. Chem., Vol. 283, Issue 8, 5158-5167, February 22, 2008

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Structural and Biophysical Analysis of the DNA Binding Properties of Myelin Transcription Factor 1^*

From the School of Molecular and Microbial Biosciences, University of Sydney, Building G08, New South Wales, Sydney 2006, Australia

Zinc binding domains, or zinc fingers (ZnFs), form one of the most numerous and most diverse superclasses of protein structural motifs in eukaryotes. Although our understanding of the functions of several classes of these domains is relatively well developed, we know much less about the molecular mechanisms of action of many others. Myelin transcription factor 1 (MyT1) type ZnFs are found in organisms as diverse as nematodes and mammals and are found in a range of sequence contexts. MyT1, one of the early transcription factors expressed in the developing central nervous system, contains seven MyT1 ZnFs that are very highly conserved both within the protein and between species. We have used a range of biophysical techniques, including NMR spectroscopy and data-driven macromolecular docking, to investigate the structural basis for the interaction between MyT1 ZnFs and DNA. Our data indicate that MyT1 ZnFs recognize the major groove of DNA in a way that appears to differ from other known zinc binding domains.

Received for publication, May 8, 2007 , and in revised form, November 27, 2007.

The atomic coordinates and structure factors (code 2JX1 and 2JYD) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

^* This work was supported in part by a National Health and Medical Research Council (NHMRC) program grant (to J. P. M. and J. M. M.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement"in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Both authors contributed equally to this work.

² Supported by a fellowship from the Austrian Science Fund (Grant J2474).

³ Supported by a PHD (plant homeodomain) scholarship from Dr. Geoff Duggin.

⁴ A Viertel Medical Research Fellow.

⁵ An NHMRC Senior Research Fellow. To whom correspondence should be addressed. Tel.: 61-2-9351-3906; Fax: 61-2-9351-4726; E-mail: j.mackay{at}mmb.usyd.edu.au.

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