Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, R, Giuseppe Máximo Scolfaro 10000, Campinas, SP 13083-970, Brazil.
BMC Genomics. 2014 Feb 25;15:157. doi: 10.1186/1471-2164-15-157.
Transcriptional activator-like (TAL) effectors, formerly known as the AvrBs3/PthA protein family, are DNA-binding effectors broadly found in Xanthomonas spp. that transactivate host genes upon injection via the bacterial type three-secretion system. Biologically relevant targets of TAL effectors, i.e. host genes whose induction is vital to establish a compatible interaction, have been reported for xanthomonads that colonize rice and pepper; however, citrus genes modulated by the TAL effectors PthA"s" and PthC"s" of the citrus canker bacteria Xanthomonas citri (Xc) and Xanthomonas aurantifolii pathotype C (XaC), respectively, are poorly characterized. Of particular interest, XaC causes canker disease in its host lemon (Citrus aurantifolia), but triggers a defense response in sweet orange.
Based on, 1) the TAL effector-DNA binding code, 2) gene expression data of Xc and XaC-infiltrated sweet orange leaves, and 3) citrus hypocotyls transformed with PthA2, PthA4 or PthC1, we have identified a collection of Citrus sinensis genes potentially targeted by Xc and XaC TAL effectors. Our results suggest that similar with other strains of Xanthomonas TAL effectors, PthA2 and PthA4, and PthC1 to some extent, functionally converge. In particular, towards induction of genes involved in the auxin and gibberellin synthesis and response, cell division, and defense response. We also present evidence indicating that the TAL effectors act as transcriptional repressors and that the best scoring predicted DNA targets of PthA"s" and PthC"s" in citrus promoters predominantly overlap with or localize near to TATA boxes of core promoters, supporting the idea that TAL effectors interact with the host basal transcriptional machinery to recruit the RNA pol II and start transcription.
The identification of PthA"s" and PthC"s" targets, such as the LOB (lateral organ boundary) and CCNBS genes that we report here, is key for the understanding of the canker symptoms development during host susceptibility, or the defenses of sweet orange against the canker bacteria. We have narrowed down candidate targets to a few, which pointed out the host metabolic pathways explored by the pathogens.
转录激活子样(TAL)效应子,以前称为 AvrBs3/PthA 蛋白家族,是广泛存在于黄单胞菌属中的 DNA 结合效应子,通过细菌的 III 型分泌系统注入后可使宿主基因转录激活。已报道了在定殖水稻和辣椒的黄单胞菌中 TAL 效应子的生物相关靶标,即诱导其建立相容互作至关重要的宿主基因;然而,由柑橘溃疡病菌 Xanthomonas citri(Xc)和柑橘溃疡病菌黄化型 C(XaC)的 PthA“s”和 PthC“s”TAL 效应子调节的柑橘基因的特征描述较差。特别值得关注的是,XaC 在其宿主柠檬(Citrus aurantifolia)中引起溃疡病,但在甜橙中触发防御反应。
基于 1)TAL 效应子-DNA 结合密码子,2)Xc 和 XaC 浸润甜橙叶片的基因表达数据,以及 3)用 PthA2、PthA4 或 PthC1 转化的柑橘下胚轴,我们鉴定了一套可能被 Xc 和 XaC TAL 效应子靶向的柑橘基因。我们的结果表明,与其他黄单胞菌菌株的 TAL 效应子类似,PthA2 和 PthA4,以及在某种程度上的 PthC1,功能上趋于一致。特别是,诱导参与生长素和赤霉素合成和反应、细胞分裂和防御反应的基因。我们还提供了证据表明,TAL 效应子作为转录抑制剂起作用,并且 PthA“s”和 PthC“s”在柑橘启动子中的最佳评分预测 DNA 靶标主要与核心启动子的 TATA 框重叠或位于其附近,支持 TAL 效应子与宿主基本转录机制相互作用以招募 RNA pol II 并启动转录的观点。
鉴定 PthA“s”和 PthC“s”的靶标,例如我们在此报告的 LOB(侧生器官边界)和 CCNBS 基因,是理解宿主易感性期间溃疡症状发展或甜橙对溃疡细菌防御的关键。我们已经将候选靶标缩小到少数几个,这些靶标指出了病原体探索的宿主代谢途径。