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ETHYLENE RESPONSE FACTOR4 的具有激活子和阻遏子功能的交替多聚腺苷酸化异构体对 L. 衰老的影响

Impact of Alternatively Polyadenylated Isoforms of ETHYLENE RESPONSE FACTOR4 with Activator and Repressor Function on Senescence in L.

机构信息

Center for Plant Molecular Biology (ZMBP), University of Tuebingen, 72076 Tuebingen, Germany.

出版信息

Genes (Basel). 2019 Jan 28;10(2):91. doi: 10.3390/genes10020091.

Abstract

Leaf senescence is highly regulated by transcriptional reprogramming, implying an important role for transcriptional regulators. ETHYLENE RESPONSE FACTOR4 (ERF4) was shown to be involved in senescence regulation and to exist in two different isoforms due to alternative polyadenylation of its pre-mRNA. One of these isoforms, ERF4-R, contains an ERF-associated amphiphilic repression (EAR) motif and acts as repressor, whereas the other form, ERF4-A, is lacking this motif and acts as activator. Here, we analyzed the impact of these isoforms on senescence. Both isoforms were able to complement the delayed senescence phenotype of the mutant with a tendency of ERF4-A for a slightly better complementation. However, overexpression led to accelerated senescence of 35S: plants but not of 35S: plants. We identified () as direct target gene of ERF4 in a yeast-one-hybrid screen. Both isoforms directly bind to the promoter but have antagonistic effects on gene expression. The ratio of to mRNA changed during development, leading to a complex age-dependent regulation of CAT3 activity. The RNA-binding protein FPA shifted the R/A-ratio and mutants are pointing towards a role of alternative polyadenylation regulators in senescence.

摘要

叶片衰老受转录重编程的高度调控,这意味着转录调控因子在其中起着重要作用。ETHYLENE RESPONSE FACTOR4(ERF4)被证明参与衰老调控,并由于其前体 mRNA 的可变多聚腺苷酸化而存在两种不同的异构体。这些异构体之一,ERF4-R,含有一个 ERF 相关的两亲性抑制(EAR)基序,作为抑制剂,而另一种形式,ERF4-A,缺乏这个基序,作为激活剂。在这里,我们分析了这些异构体对衰老的影响。两种异构体都能够补充 的延迟衰老表型,而 ERF4-A 则具有稍微更好的补充作用。然而,过表达导致 35S:植物加速衰老,但 35S:植物没有。我们在酵母单杂交筛选中鉴定出 ()是 ERF4 的直接靶基因。两种异构体都直接结合到 启动子上,但对基因表达有拮抗作用。在发育过程中, 到 的 mRNA 比率发生变化,导致 CAT3 活性的复杂年龄依赖性调控。RNA 结合蛋白 FPA 改变了 R/A 比,而 突变体则表明可变多聚腺苷酸化调节剂在衰老中起作用。

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