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Nucleic Acids Research, 1994, Vol. 22, No. 24 5164-5172
© 1994

Articles

Direct analysis of native and chimeric GATA specific DNA binding proteins from Aspergillus nidulans

David G. Peters and Mark X. Caddick*

Department of Genetics and Microbiology, Donnan Laboratories, University of Liverpool PO Box 147, Liverpool, L69 3BX, UK

*To whom correspondence should be addressed

Received October 18, 1994. Accepted November 2, 1994.

In Aspergillus nidulans the regulatory gene areA Is responsible for mediating nitrogen metabolite repression. The areA product (AREA) represents an example of the GATA family of DNA binding proteins, which are characterised by the presence of a GATA domain consisting of a zinc finger within a highly conserved region of 52 amino acids. Among the other transcription factors Included in this family Is the principal erythrold transcription factor, GATA-1, which contains two GATA domains. In order to demonstrate high specificity binding of native AREA to DNA containing the sequence -GATA-, and investigate the presence in A.nidulans of other proteins with related specificities, we have used gel mobility shift assays. Both AREA-dependent and independent complexes have been identified. Two strains bearing chimeric genes were also characterised. In these, the region encoding the native GATA domain of AREA was replaced by sequences from murtne GATA-1 cDNA encoding either the equivalent C-termlnal domain or both the N and C-terminal domains. Strains bearing the areA::NC-GATA construct, which Includes the sequence encoding both the N and C-terminal domains of GATA–1, leads to a pronounced Increase in one of two AREA-dependent complexes and Implicates the N-termlnal domain of GATA-1 in mediating protein-protein interactions.


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