Snoeijers S S, Vossen P, Goosen T, Van den Broek H W, De Wit P J
Wageningen Agricultural University, Laboratory of Genetics, The Netherlands.
Mol Gen Genet. 1999 Jun;261(4-5):653-9. doi: 10.1007/s004380050008.
The avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum is highly induced during infection of tomato plants. Expression of the Avr9 gene can also be induced in vitro when cells are grown on synthetic liquid medium containing little or no nitrogen. The Avr9 promoter contains six copies of the sequence TAGATA and six additional copies of the core sequence GATA within 0.4 kb upstream of the translation start site. In the filamentous fungi Aspergillus nidulans and Neurospora crassa, these promoter sequences have been identified as the binding sites for a wide-domain GATA-type regulator (AREA in A. nidulans and NIT2 in N. crassa) involved in nitrogen utilization. Quantification of GUS activity of A. nidulans transformants containing a single copy of the fully active Avr9 promoter-uidA (GUS) reporter gene fusion in different areA backgrounds, following starvation for nitrogen, showed that induction of the Avr9 promoter is regulated similarly in A. nidulans and C. fulvum. This suggests that AREA can regulate the Avr9 promoter and that C. fulvum contains an AREA-like regulator that can bind to these specific sequence motifs. Comparison of the induction profiles of Avr9 and niaD showed that Avr9 expression is independent of NIRA, as is niaD expression upon nitrogen starvation. Studies with Avr9 promoter-uidA fusions in which all or most of these sequences had been deleted, showed that Avr9 promoter activity is dependent on the presence of these specific cis-regulatory elements, suggesting that they do indeed function in transcriptional regulation of the Avr9 gene.
番茄真菌病原体fulvum的无毒基因Avr9在感染番茄植株的过程中被高度诱导。当细胞在含很少或不含氮的合成液体培养基上生长时,Avr9基因的表达也可在体外被诱导。Avr9启动子在翻译起始位点上游0.4 kb范围内含有6个TAGATA序列拷贝和6个额外的核心序列GATA拷贝。在丝状真菌构巢曲霉和粗糙脉孢菌中,这些启动子序列已被鉴定为参与氮利用的广泛结构域GATA型调节因子(构巢曲霉中的AREA和粗糙脉孢菌中的NIT2)的结合位点。对在不同areA背景下含有单拷贝完全活性Avr9启动子-uidA(GUS)报告基因融合体的构巢曲霉转化体进行氮饥饿处理后,对GUS活性进行定量分析,结果表明Avr9启动子的诱导在构巢曲霉和fulvum中受到类似的调节。这表明AREA可以调节Avr9启动子,并且fulvum含有一种类似AREA的调节因子,它可以与这些特定的序列基序结合。对Avr9和niaD诱导谱的比较表明,Avr9的表达独立于NIRA,氮饥饿时niaD的表达也是如此。对其中所有或大部分这些序列已被删除的Avr9启动子-uidA融合体的研究表明,Avr9启动子活性依赖于这些特定顺式调节元件的存在,这表明它们确实在Avr9基因的转录调控中发挥作用。