The regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. Mutations affecting specificity of gene activation alter a loop residue of a putative zinc finger.
The regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans has been sequenced and its transcript mapped and orientated. A single ORF can encode a protein of 719 amino acids. A 52 amino acid region including a putative 'zinc finger' strongly resembles putative DNA binding regions of the major regulatory protein of erythroid cells. The derived protein sequence also contains a highly acidic region possibly involved in gene activation and 22 copies of the motif S(T)PXX, abundant in DNA binding proteins. Analysis of chromosomal rearrangements and transformation with deletion clones identified 342 N-terminal and 124 C-terminal residues as inessential and localized a C-terminal region required for nitrogen metabolite repressibility. A -1 frameshift eliminating the inessential 122 C-terminal amino acids is a surprising loss-of-function mutation. Extraordinary basicity of the replacement C terminus might explain its phenotype. Mutant sequencing also identified a polypeptide chain termination and several missense mutations, but most interesting are sequence changes associated with specificity mutations. A mutation elevating expression of some structural genes under areA control whilst reducing or not affecting expression of others is a leucine to valine change in the zinc finger loop. It reverts to a partly reciprocal phenotype by replacing the mutant valine by methionine.
Kudla
B
B
Department of Bacteriology, Royal Postgraduate Medical School, London, UK.
Caddick
M X
MX
Langdon
T
T
Martinez-Rossi
N M
NM
Bennett
C F
CF
Sibley
S
S
Davies
R W
RW
Arst
H N
HN
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics*; D001483:Base Sequence; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000235:genetics*; D050981:Erythroid-Specific DNA-Binding Factors; D005656:Fungal Proteins / Q000235:genetics*; D015321:Gene Rearrangement; D005800:Genes, Fungal; D005809:Genes, Regulator; D008667:Metalloproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism*; D011061:Poly A / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors; D015032:Zinc
D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D005656:Fungal Proteins; D008667:Metalloproteins; D012333:RNA, Messenger; D014157:Transcription Factors; D011061:Poly A; D015032:Zinc; D009584:Nitrogen