Saleeba J A, Cobbett C S, Hynes M J
Department of Genetics, University of Melbourne, Parkville, Victoria, Australia.
Mol Gen Genet. 1992 Nov;235(2-3):349-58. doi: 10.1007/BF00279380.
The structure, function and regulation of the acetate inducible aciA gene of Aspergillus nidulans was analysed. The aciA locus was mapped to chromosome 1 at a position where no acetate inducible gene has been previously located. The nucleotide sequence of aciA was determined, the structures of two transcripts were determined and the sequences of the polypeptide products of the gene were deduced. Construction of an aciA loss-of-function mutant was achieved via insertional inactivation, but it did not reveal a phenotype for an aciA- strain. The larger polypeptide, AciA, was found to have a putative dinucleotide cofactor binding site. Acetate inducibility of aciA was found to be dependent on the amdA regulatory gene. Use of a 5' deletion series of an aciA--lacZ fusion and an in vivo regulatory protein binding (titration) assay allowed the region required for amdA activity to be localized to a 124 bp segment 5' to aciA. A 13 bp region of sequence similarity was observed between aciA and the coregulated amdS gene in the regions required for amdA-mediated regulation of these genes. This sequence may have a role in amdA regulation of the two genes.
对构巢曲霉中乙酸盐诱导型aciA基因的结构、功能及调控进行了分析。aciA基因座被定位到1号染色体上一个先前未定位过乙酸盐诱导型基因的位置。测定了aciA的核苷酸序列,确定了两种转录本的结构,并推导了该基因多肽产物的序列。通过插入失活构建了aciA功能缺失突变体,但未发现aciA-菌株的表型。发现较大的多肽AciA具有一个假定的二核苷酸辅因子结合位点。发现aciA的乙酸盐诱导性依赖于amdA调控基因。使用aciA-lacZ融合的5'缺失系列和体内调控蛋白结合(滴定)试验,可将amdA活性所需区域定位到aciA 5'端的一个124 bp片段。在aciA与共调控的amdS基因之间,在amdA介导的这些基因调控所需区域观察到13 bp的序列相似性。该序列可能在amdA对这两个基因的调控中起作用。