Tag A G, Garifullina G F, Peplow A W, Ake C, Phillips T D, Hohn T M, Beremand M N
Department of Plant Pathology and Microbiology, Texas A&M University, College Station, Texas 77843, USA.
Appl Environ Microbiol. 2001 Nov;67(11):5294-302. doi: 10.1128/AEM.67.11.5294-5302.2001.
We report here the characterization of Tri10, a novel regulatory gene within the trichothecene gene cluster. Comparison of Tri10 genomic and mRNA sequences revealed that removal of a single 77-bp intron provided a 1,260-bp open reading frame, encoding a 420-amino-acid protein. Disruption of Tri10 in Fusarium sporotrichioides abolished T-2 toxin production and dramatically decreased the transcript accumulation for four trichothecene genes (Tri4, Tri5, Tri6, and Tri101) and an apparent farnesyl pyrophosphate synthetase (Fpps) gene. Conversely, homologous integration of a disruption vector by a single upstream crossover event significantly increased T-2 toxin production and elevated the transcript accumulation of the trichothecene genes and Fpps. Further analysis revealed that disruption of Tri10, and to a greater extent the disruption of Tri6, increased sensitivity to T-2 toxin under certain growth conditions. Although Tri10 is conserved in Fusarium graminearum and Fusarium sambucinum and clearly plays a central role in regulating trichothecene gene expression, it does not show any significant matches to proteins of known or predicted function or to motifs except a single transmembrane domain. We suggest a model in which Tri10 acts upstream of the cluster-encoded transcription factor TRI6 and is necessary for full expression of both the other trichothecene genes and the genes for the primary metabolic pathway that precedes the trichothecene biosynthetic pathway, as well as for wild-type levels of trichothecene self-protection. We further suggest the presence of a regulatory loop where Tri6 is not required for the transcription of Tri10 but is required to limit the expression of Tri10.
我们在此报告了Tri10的特性,它是单端孢霉烯基因簇中的一个新型调控基因。Tri10基因组序列与mRNA序列的比较显示,去除一个77碱基对的单一内含子后产生了一个1260碱基对的开放阅读框,编码一个420个氨基酸的蛋白质。在拟分枝镰刀菌中破坏Tri10消除了T-2毒素的产生,并显著降低了四个单端孢霉烯基因(Tri4、Tri5、Tri6和Tri101)以及一个明显的法尼基焦磷酸合成酶(Fpps)基因的转录积累。相反,通过单个上游交叉事件进行破坏载体的同源整合显著增加了T-2毒素的产生,并提高了单端孢霉烯基因和Fpps的转录积累。进一步分析表明,在某些生长条件下,破坏Tri10,以及在更大程度上破坏Tri6,会增加对T-2毒素的敏感性。尽管Tri10在禾谷镰刀菌和接骨木镰刀菌中保守,并且在调节单端孢霉烯基因表达中显然起着核心作用,但除了一个单一的跨膜结构域外,它与已知或预测功能的蛋白质或基序没有任何显著匹配。我们提出了一个模型,其中Tri10在簇编码的转录因子TRI6的上游起作用,对于其他单端孢霉烯基因以及单端孢霉烯生物合成途径之前的初级代谢途径基因的完全表达,以及对于单端孢霉烯自我保护的野生型水平而言是必需的。我们进一步提出存在一个调控环,其中Tri6对于Tri10的转录不是必需的,但对于限制Tri10的表达是必需的。