amdR product and a CCAAT-binding factor bind to adjacent, possibly overlapping DNA sequences in the promoter region of the Aspergillus nidulans amdS gene

The amdS gene of Aspergillus nidulans is regulated by a number of positively acting regulatory genes which act additively and independently. Using gel mobility shift assays with crude nuclear extracts we show here that the product of one of these regulatory genes, the amdR gene, binds to DNA fragmen...

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Bibliographic Details
Published in:Nucleic acids research 1991-05, Vol.19 (10), p.2655-2660
Main Authors: Van Heeswijck, R, Hynes, M.J
Format: Article
Language:English
Subjects:
Amidohydrolases - genetics
Aspergillus nidulans
Aspergillus nidulans - enzymology
Aspergillus nidulans - genetics
Aspergillus nidulellus
Base Sequence
Binding Sites
Binding, Competitive
Biological and medical sciences
CCAAT-Enhancer-Binding Proteins
DNA, Fungal - metabolism
DNA-binding proteins
DNA-Binding Proteins - metabolism
Fundamental and applied biological sciences. Psychology
Fungal Proteins - metabolism
Genes, Fungal
genetic regulation
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
nucleotide sequences
Oligonucleotide Probes
Promoter Regions, Genetic
transcription (genetics)
Transcription. Transcription factor. Splicing. Rna processing
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Summary:The amdS gene of Aspergillus nidulans is regulated by a number of positively acting regulatory genes which act additively and independently. Using gel mobility shift assays with crude nuclear extracts we show here that the product of one of these regulatory genes, the amdR gene, binds to DNA fragments containing part of the promoter region of the amdS gene. This confirms the earlier prediction from DNA sequence data that amdR encodes a DNA-binding protein containing a cysteine-rich 'zinc finger' motif. In addition we detected the binding of another previously unidentified protein to an adjacent, possibly overlapping region of the amdS 51 sequence at the site of a consensus 'CCAAT-box' sequence. Replacement of the CCAAT sequence with CCTTT abolished the binding of this protein which we have designated as an A. nidulans 'CCAAT-box' binding factor (AnCF). The 'CCAAT-box' sequence appears to be involved in determining the basal level of transcription of amdS (T.G. Littlejohn and M.J.H., unpublished data). This suggests that AnCF is a transcription factor, and that the 'CCAAT-box' sequences found in the promoters of some filamentous fungal genes function as binding sites for these factors, as in other eucaryotes.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/19.10.2655