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- Title
Mutational analysis reveals dispensability of the N-terminal region of the Aspergillus transcription factor mediating nitrogen metabolite repression.
- Authors
Langdon, Tim; Sheerins, Alan; Ravagnani, Adriana; Gielkens, Marco; Caddick, Mark X.; Arst Jr., Herbert N.
- Abstract
Mutational analysis has enabled identification and localization of an upstream exon of the <em>areA</em> gene of <em>Aspergillus nidulans</em> mediating nitrogen metabolite repression. A mutation in the initiation codon and frameshift mutations, which revert by restoration of the reading frame, established the coding role of the exon and mutations affecting intron splicing in conjunction with DNA sequencing of reverse transcriptase polymerase chain reaction (RT-PCR) products localized the coding region intron. The resulting AREA translation product would have 876 residues. Deletion of the upstream exon such that translation of the remaining <em>areA</em> coding region would yield a protein containing only the 719 C-terminal residues has only a subtle phenotype, very similar to those resulting from single amino acid replacements in upstream exon-encoded regions of strong sequence similarity to the <em>Neurospora crassa</em> and <em>Penicillium chrysogenum</em> homologues. A number of <em>areA</em> mRNAs of different sizes are synthesised and appear to be functionally redundant. Synthesis of at least the smallest mRNA(s) is probably subject to autogenous activation. Suppression of frameshift mutations by compensating mutations preventing intron splicing suggests that insertion of a markedly hydrophobic sequence can impair AREA function. Finally, translational initiation for <em>areA</em> can occur within a region of at least 123 nucleotides.
- Subjects
GENETIC mutation; ASPERGILLUS nidulans; DNA polymerases; MESSENGER RNA; NUCLEOTIDE sequence
- Publication
Molecular Microbiology, 1995, Vol 17, Issue 5, p877
- ISSN
0950-382X
- Publication type
Article
- DOI
10.1111/j.1365-2958.1995.mmi_17050877.x