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Originally published In Press as doi:10.1074/jbc.M106256200 on July 9, 2001

J. Biol. Chem., Vol. 276, Issue 38, 35794-35801, September 21, 2001
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The N-terminal Zinc Finger of the Erythroid Transcription Factor GATA-1 Binds GATC Motifs in DNA*

Anthea NewtonDagger , Joel Mackay, and Merlin Crossley§

From the Department of Biochemistry, G08, University of Sydney, New South Wales, Australia 2006

The mammalian transcription factor GATA-1 is required for normal erythroid and megakaryocytic development. GATA-1 contains two zinc fingers, the C-terminal finger, which is known to bind (A/T)GATA(A/G) motifs in DNA and the N-finger, which is important for interacting with co-regulatory proteins such as Friend of GATA (FOG). We now show that, like the C-finger, the N-finger of GATA-1 is also capable of binding DNA but recognizes distinct sequences with the core GATC. We demonstrate that the GATA-1 N-finger can bind these sequences in vitro and that in cellular assays, GATA-1 can activate promoters containing GATC motifs. Experiments with mutant GATA-1 proteins confirm the importance of the N-finger, as the C-finger is not required for transactivation from GATC sites. Recently four naturally occurring mutations in GATA-1 have been shown to be associated with familial blood disorders. These mutations all map to the N-finger domain. We have investigated the effect of these mutations on the recognition of GATC sites by the N-finger and show that one mutation R216Q abolishes DNA binding, whereas the others have only minor effects.

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

Dagger Supported by an Australian Postgraduate Award.

§ To whom correspondence should be addressed: Dept. of Biochemistry, G08, University of Sydney, NSW, Australia, 2006. Tel.: 61 2 9351 2233; Fax: 61 2 9351 4726; E-mail: M.Crossley@biochem.usyd.edu.au.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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