Conservation and Diversity of MicroRNA-associated Copper-regulatory Networks in Populus trichocarpa

Conservation and Diversity of MicroRNA-associated Copper-regulatory Networks in Populus trichocarpa
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毛果杨中 microRNA 相关铜调节网络的保守性和多样性。

DOI:
10.1111/j.1744-7909.2011.01080.x
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发表时间:
2011-11-01
影响因子:
11.4
通讯作者:
Chiang, Vincent L.
Chiang, Vincent L.
中科院分区:
生物学1区
文献类型:
--
作者:
Lu, Shanfa;Yang, Chenmin;Chiang, Vincent L.

文献摘要

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工厂开发重要的监管网络以适应铜 (Cu) 供应量的频繁变化。然而,人们对拟南芥以外的植物物种中与 miRNA 相关的铜调控网络知之甚少。在这里,我们报道毛果杨 (Torr. & Gray) 中的 Cu 响应 miRNA 不仅包括保守的 miR397、miR398 和 miR408,还包括杨属特异性 miR1444,表明植物中 Cu 响应 miRNA 的保守性和多样性。与铜相关的成熟 miRNA 的抑制与杨属中编码含铜蛋白的靶基因的上调相伴随。这些靶标包括 miR397 靶向的 PtLAC5、PtLAC6 和 PtLAC110a,miR398 靶向的 PtCSD1、PtCSD2a 和 PtCSD2b,miR408 靶向的 PtPCL1、PtPCL2、PtPCL3 和 PtLAC4,以及 miR1444 靶向的 PtPPO3 和 PtPPO6。一致地,转基因烟草植物中 Cu 下调毛果 P. trichocarpa miR408 启动子指导的 GUS 基因表达。 Cu 响应元件 (CuRE) 存在于 Cu 响应 miRNA 基因的启动子中。我们鉴定了 34 个 SQUAMOSA 启动子结合蛋白样 (SPL) 基因,其中 17 个是全长 PtSPL 蛋白或具有至少 300 个氨基酸的部分序列。系统发育分析表明 PtSPL3 和 PtSPL4 是控制 Cu 响应基因表达的 CuRE 结合蛋白。 Cu 似乎不参与这些转录因子的调节,因为 PtSPL3 和 PtSPL4 都不受 Cu 调节,并且它们的启动子中不存在 CuRE。
Plants develop important regulatory networks to adapt to the frequently-changing availability of copper (Cu). However, little is known about miRNA-associated Cu-regulatory networks in plant species other than Arabidopsis. Here, we report that Cu-responsive miRNAs in Populus trichocarpa (Torr. & Gray) include not only conserved miR397, miR398 and miR408, but also Populus-specific miR1444, suggesting the conservation and diversity of Cu-responsive miRNAs in plants. Copper-associated suppression of mature miRNAs is in company with the up-regulation of their target genes encoding Cu-containing proteins in Populus. The targets include miR397-targeted PtLAC5, PtLAC6 and PtLAC110a, miR398-targeted PtCSD1, PtCSD2a and PtCSD2b, miR408-targeted PtPCL1, PtPCL2, PtPCL3 and PtLAC4, and miR1444-targeted PtPPO3 and PtPPO6. Consistently, P. trichocarpa miR408 promoter-directed GUS gene expression is down-regulated by Cu in transgenic tobacco plants. Cu-response elements (CuREs) are found in the promoters of Cu-responsive miRNA genes. We identified 34 SQUAMOSA-promoter binding protein-like (SPL) genes, of which 17 are full-length PtSPL proteins or partial sequences with at least 300 amino acids. Phylogenetic analysis indicates that PtSPL3 and PtSPL4 are CuRE-binding proteins controlling Cu-responsive gene expression. Cu appears to be not involved in the regulation of these transcription factors because neither PtSPL3 nor PtSPL4 is Cu-regulated and no CuRE exists in their promoters.