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Molecular and Cellular Biology, January 1999, p. 99-106, Vol. 19, No. 1
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

AnCF, the CCAAT Binding Complex of Aspergillus nidulans, Contains Products of the hapB, hapC, and hapE Genes and Is Required for Activation by the Pathway-Specific Regulatory Gene amdR

Stefan Steidl,1,2,dagger Peter Papagiannopoulos,1 Olivier Litzka,2 Alex Andrianopoulos,1 Meryl A. Davis,1 Axel A. Brakhage,2,dagger and Michael J. Hynes1,*

Department of Genetics, University of Melbourne, Parkville, Victoria 3052, Australia,1 and Institut fuer Genetik und Mikrobiologie, Ludwig-Maximilians-Universität, Munich 80638, Germany2

Received 2 July 1998/Returned for modification 25 August 1998/Accepted 29 September 1998

CCAAT binding factors (CBFs) positively regulating the expression of the amdS gene (encoding acetamidase) and two penicillin biosynthesis genes (ipnA and aatA) have been previously found in Aspergillus nidulans. The factors were called AnCF and PENR1, respectively. Deletion of the hapC gene, encoding a protein with significant similarity to Hap3p of Saccharomyces cerevisiae, eliminated both AnCF and PENR1 binding activities. We now report the isolation of the genes hapB and hapE, which encode proteins with central regions of high similarity to Hap2p and Hap5p of S. cerevisiae and to the CBF-B and CBF-C proteins of mammals. An additional fungus-specific domain present in HapE was revealed by comparisons with the homologs from S. cerevisiae, Neurospora crassa, and Schizosaccharomyces pombe. The HapB, HapC, and HapE proteins have been shown to be necessary and sufficient for the formation of a CCAAT binding complex in vitro. Strains with deletions of each of the hapB, hapC, and hapE genes have identical phenotypes of slow growth, poor conidiation, and reduced expression of amdS. Furthermore, induction of amdS by omega amino acids, which is mediated by the AmdR pathway-specific activator, is abolished in the hap deletion mutants, as is growth on gamma -aminobutyric acid as a sole nitrogen or carbon source. AmdR and AnCF bind to overlapping sites in the promoters of the amdS and gatA genes. It is known that AnCF can bind independently of AmdR. We suggest that AnCF binding is required for AmdR binding in vivo.

* Corresponding author. Mailing address: Department of Genetics, University of Melbourne, Parkville, Victoria 3052, Australia. Phone: (61 3) 9344 6246. Fax: (61 3) 9344 5139. E-mail: hynes.lab{at}genetics.unimelb.edu.au.

dagger Present address: Institut für Mikrobiologie und Genetik, Technische Universität Darmstadt, 64287 Darmstadt, Germany.

Molecular and Cellular Biology, January 1999, p. 99-106, Vol. 19, No. 1
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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