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Molecular and Cellular Biology, October 1999, p. 6523-6531, Vol. 19, No. 10
Department of Genetics, University of
Melbourne, Parkville, Victoria 3052, Australia
Received 18 March 1999/Returned for modification 28 April
1999/Accepted 17 June 1999
The CCAAT sequence in the amdS promoter of
Aspergillus nidulans is recognized by AnCF, a complex
consisting of the three evolutionary conserved subunits HapB, HapC, and
HapE. In this study we have investigated the effect of AnCF on the
chromatin structure of the amdS gene. The AnCF complex and
the CCAAT sequence were found to be necessary for the formation of a
nucleosome-free, DNase I-hypersensitive region in the 5' region of the
amdS gene. Deletion of the hapE gene results in
loss of the DNase I-hypersensitive site, and the positioning of
nucleosomes over the transcriptional start point is lost. Likewise, a
point mutation in the CCAAT motif, as well as a 530-bp deletion which
removes the CCAAT box, results in the loss of the DNase
I-hypersensitive region. The DNase I-hypersensitive region and the
nucleosome positioning can be restored by insertion of a 35-bp
oligonucleotide carrying the CCAAT motif. A DNase I-hypersensitive region has been found in the CCAAT-containing fmdS gene and
was also hapE dependent. These data indicate a critical
role for the AnCF complex in establishing an open chromatin structure
in A. nidulans.
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
AnCF, the CCAAT Binding Complex of
Aspergillus nidulans, Is Essential for the Formation of a
DNase I-Hypersensitive Site in the 5' Region of the
amdS Gene
*
Corresponding author. Mailing address: Department of
Genetics, University of Melbourne, Gate 12, Royal Parade Parkville,
Victoria 3052, Australia. Phone: (61) (3) 9344-6246. Fax: (61) (3)
9344-5139. E-mail:
hynes.lab{at}genetics.unimelb.edu.au.
Molecular and Cellular Biology, October 1999, p. 6523-6531, Vol. 19, No. 10
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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