MUTATIONAL ANALYSIS REVEALS DISPENSABILITY OF THE N-TERMINAL REGION OF THE ASPERGILLUS TRANSCRIPTION FACTOR MEDIATING NITROGEN METABOLITE REPRESSION

被引:79
作者
LANGDON, T
SHEERINS, A
RAVAGNANI, A
GIELKENS, M
CADDICK, MX
ARST, HN
机构
[1] ROYAL POSTGRAD MED SCH,DEPT INFECT DIS & BACTERIOL,LONDON W12 0NN,ENGLAND
[2] UNIV LIVERPOOL,DEPT GENET & MICROBIOL,DONNAN LABS,LIVERPOOL L69 3BX,MERSEYSIDE,ENGLAND
关键词
D O I
10.1111/j.1365-2958.1995.mmi_17050877.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Mutational analysis has enabled identification and localization of an upstream exon of the areA gene of Aspergillus nidulans 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 areA 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 Neurospora crassa and Penicillium chrysogenum homologues. A number of areA 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 areA can occur within a region of at least 123 nucleotides.
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收藏
页码:877 / 888
页数:12
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