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aroG1和aroG2两种假定的DAHP合酶的功能表征及其与Ⅲ型分泌系统的联系

Functional Characterization of Two Putative DAHP Synthases of AroG1 and AroG2 and Their Links With Type III Secretion System in .

作者信息

Zhang Weiqi, Li Jing, Shi Xiaojun, Hikichi Yasufumi, Zhang Yong, Ohnishi Kouhei

机构信息

College of Resources and Environment, Southwest University, Chongqing, China.

The Ninth People's Hospital of Chongqing, Chongqing, China.

出版信息

Front Microbiol. 2019 Feb 12;10:183. doi: 10.3389/fmicb.2019.00183. eCollection 2019.

Abstract

Type three secretion system (T3SS) is essential for to cause disease in host plants and we previously screened AroG1 as a candidate with impact on the T3SS expression. Here, we focused on two putative DAHP synthases of AroG1 and AroG2, which control the first step of the shikimate pathway, a common route for biosynthesis of aromatic amino acids (AAA), to characterize their functional roles and possible links with virulence in . Deletion of or , but not , significantly impaired the T3SS expression both and , and the impact of AroG1 on T3SS was mediated with a well-characterized PrhA signaling cascade. Virulence of the or mutants was completely diminished or significantly impaired in tomato and tobacco plants, but not the mutants. The mutants failed to grow in limited medium, but grew slowly . This significantly impaired growth was also observed in the mutants both and limited medium, but not in mutants. Complementary significantly restored the impaired or diminished bacterial growth, T3SS expression and virulence. Supplementary AAA or shikimic acid, an important intermediate of the shikimate pathway, significantly restored diminished growth in limited medium. The promoter activity assay showed that expression of and was greatly increased to 10-20-folder higher levels with deletion of the other. All these results demonstrated that both AroG1 and AroG2 are involved in the shikimate pathway and cooperatively essential for AAA biosynthesis in . The AroG1 plays a major role on bacterial growth, T3SS expression and pathogenicity, while the AroG2 is capable to partially carry out the function of AroG1 in the absence of AroG1.

摘要

三型分泌系统(T3SS)对于在宿主植物中致病至关重要,我们之前筛选出AroG1作为对T3SS表达有影响的候选基因。在此,我们聚焦于AroG1和AroG2这两个假定的3-脱氧-D-阿拉伯庚酮糖-7-磷酸(DAHP)合酶,它们控制莽草酸途径的第一步,莽草酸途径是芳香族氨基酸(AAA)生物合成的常见途径,以表征它们的功能作用以及与[具体对象]毒力的可能联系。缺失aroG1或aroG2,但不包括aroG3,在[具体对象]和[具体对象]中均显著损害T3SS表达,并且AroG1对T3SS的影响是通过一个特征明确的PrhA信号级联介导的。aroG1或aroG2突变体在番茄和烟草植物中的毒力完全丧失或显著受损,但aroG3突变体没有。aroG1和aroG2突变体在有限培养基中无法生长,但在[具体条件]下生长缓慢。在[具体条件]和有限培养基中,aroG1和aroG2突变体均观察到这种显著受损的生长情况,但aroG3突变体没有。互补[具体操作]显著恢复了受损或减弱的细菌生长、T3SS表达和毒力。补充AAA或莽草酸(莽草酸途径的一种重要中间体)显著恢复了在有限培养基中减弱的生长。启动子活性分析表明,缺失另一个基因时,aroG1和aroG2的表达大幅增加至高出10 - 20倍的水平。所有这些结果表明,AroG1和AroG2都参与莽草酸途径,并且对[具体对象]中AAA的生物合成是协同必需的。AroG1在细菌生长、T3SS表达和致病性方面起主要作用,而AroG2在没有AroG1的情况下能够部分执行AroG1的功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c999/6379268/e2debe7e2173/fmicb-10-00183-g001.jpg

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