nirA, the pathway-specific regulatory gene of nitrate assimilation in Aspergillus nidulans, encodes a putative GAL4-type zinc finger protein and contains four introns in highly conserved regions.
Open Access
- 1 November 1991
- journal article
- research article
- Published by Taylor & Francis in Molecular and Cellular Biology
- Vol. 11 (11) , 5746-5755
- https://doi.org/10.1128/mcb.11.11.5746
Abstract
The nucleotide sequence of nirA, mediating nitrate induction in Aspergillus nidulans, has been determined. Alignment of the cDNA and the genomic DNA sequence indicates that the gene contains four introns and encodes a protein of 892 amino acids. The deduced NIRA protein displays all characteristics of a transcriptional activator. A putative double-stranded DNA-binding domain in the amino-terminal part comprises six cysteine residues, characteristic for the GAL4 family of zinc finger proteins. An amino-terminal highly acidic region and two proline-rich regions are also present. The nucleotide sequences of two mutations were determined after they were mapped by transformation with overlapping DNA fragments, amplified by the polymerase chain reaction. nirA87, a mutation conferring noninducibility by nitrate and nitrite, has a -1 frameshift at triplet 340, which eliminates 549 C-terminal amino acids from the polypeptide. Under the assumption that the truncated polypeptide is stable, it comprises the zinc finger domain and the acidic region, which seem not sufficient for transcriptional activation. nirAd-106, an allele conferring nitrogen metabolite derepression of nitrate and nitrite reductase activity, includes two transitions, changing a glutamic acid to a lysine and a valine to an alanine, situated between a basic and a proline-rich region of the protein. Northern (RNA) analysis of the wild type and of constitutive (nirAc) and derepressed (nirAd) mutants show that the nirA transcript does not vary between these strains, being in all cases constitutively expressed. On the other hand, transcript levels of structural genes (niaD and niiA) do vary, being highly inducible in the wild type but constitutively expressed in the nirAc mutant. The nirAd mutant appears phenotypically derepressed, because the niaD and niiA transcript levels are overinduced in the presence of nitrate but are still partially repressed in the presence of ammonium.Keywords
This publication has 36 references indexed in Scilit:
- Correct intron splicing generates a new type of a putative zinc‐binding domain in a transcriptional activator of Aspergillus nidulansFEBS Letters, 1991
- The proline-rich transcriptional activator of CTF/NF-I is distinct from the replication and DNA binding domainCell, 1989
- Transcriptional Regulation in Mammalian Cells by Sequence-Specific DNA Binding ProteinsScience, 1989
- DNA binding specificity of steroid receptorsCell, 1989
- SPXX, a frequent sequence motif in gene regulatory proteinsJournal of Molecular Biology, 1989
- Helix-turn-helix, zinc-finger, and leucine-zipper motifs for eukaryotic transcriptional regulatory proteinsTrends in Biochemical Sciences, 1989
- Functional dissection and sequence of yeast HAP1 activatorCell, 1989
- Intron within the large rRNA gene of N. crassa mitochondria: A long open reading frame and a consensus sequence possibly important in splicingCell, 1982
- Mutations in nirA gene of Aspergillus nidulans and nitrogen metabolismNature, 1978
- Regulation of Nitrate Reduction in Aspergillus nidulansNature, 1967