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A Proximal Promoter Region of Arabidopsis DREB2C Confers Tissue-specific Expression under Heat Stress

Journal of Integrative Plant Biology 2012年第9期54卷 640-651页

作者： Jihyun Je Chieun Song Jung Eun Hwang Chae Oh Lim systems and synthetic agrobiotech center and pmbbrc Division of Applied Life Science Gyeongsang National University

The dehydration-responsive element-binding factor 2C （DREB2C） is a member of the CBF/DREB subfamily of proteins, which contains a single APETALA2/Ethylene responsive element-binding factor （AP2/ERF） domain. To identify the expression pattern of the DREB2C gene, which contains multiple transcription cis-regulatory elements in its promoter, an approximately 1.4 kb upstream DREB2C sequence was fused to the β-glucuronidase reporter gene （GUS） and the recombinant p1244 construct was transformed into Arabidopsis thaliana （L.） Heynh. The promoter of the gene directed prominent GUS activity in the vasculature in diverse young dividing tissues. Upon applying heat stress （HS）, GUS staining was also enhanced in the vasculature of the growing tissues. Analysis of a series of 5＇-deletions of the DREB2C promoter revealed that a proximal upstream sequence sufficient for the tissue-specific spatial and temporal induction of GUS expression by HS is localized in the promoter region between -204 and -34 bps relative to the transcriptional start site. Furthermore, electrophoretic mobility shift assay （EMSA） demonstrated that nuclear protein binding activities specific to a -120 to -32 bp promoter fragment increased after HS. These results indicate that the TATA-proximal region and some latent trans-acting factors may cooperate in HS-induced activation of the Arabidopsis DREB2C promoter.

关键词： cis-acting elements β-glucuronidase gene expression histochemical assay promoter deletion transgenic plant.

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Light-stabilized FHA2 suppresses miRNA biogenesis through interactions with DCL1 and HYL1

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Molecular Plant 2021年第4期14卷 647-663页

作者： Seung Jun Park Suk Won Choi Gu Min Kim Christian Moller Hyun-Sook Pai Seong Wook Yang Department of systems Biology College of Life Science and BiotechnologyYonsei UniversitySeoul 120749Korea EDepartment of systems Biology Institute of Life Science and BiotechnologyYonsei UniversitySeoul 120-749Korea Department of Plant and Environnental Sciences Faculty of ScienceUniversity of CopenhagenThorvaldsensvei 40DK-1871 FrederiksbergCopenhagenDenmark Pohang University of Science and Technology(POSTECH) 77 Cheongam-RoNam-GuPohangGyeongbuk37673.Korea These authors contributed equally to this article.

The precise regulation of microRNA(miRNA)biogenesis is crucial for plant development,which requires core microprocessors and many fine tuners to coordinate their miRNA processing activity/specificity in fluctuating cellular *** de-etiolation,light triggers a dramatic accumulation of core microprocessors and primary miRNAs(pri-miRNAs)but decreases pri-miRNA processing activity,resulting in relatively constant miRNA *** mechanisms underlying these seemingly contradictory regulatory changes remain *** this study,we identified forkhead-associated domain 2(FHA2)as a light-stabilized suppressor of miRNA *** found that FHA2 deficiency increased the level of mature miRNAs,accompanied by a reduction in pri-miRNAs and target *** assays showed that FHA2 associates with the core microprocessors DCL1,HYL1,and SE,forming a complex to suppress their pri-miRNA processing *** analyses revealed that FHA2 promotes HYL1 binding but inhibits the binding of DCL1-PAZ-RNase-RNA-binding domains(DCL1-PRR)to miRNAs,whereas FHA2 does not directly bind to these ***,we found that FHA2 protein is unstable in the dark but stabilized by light during ***,disruption of FH A led to defects in light-triggered changes in miRNA expression and reduced the survival rate of deetiolated seedlings after prolonged light ***,these data suggest that FHA2 is a novel light-stabilized suppressor of miRNA biogenesis and plays a role in fine-tuning miRNA processing during de-etiolation.

关键词： miRNA biogenesis Suppressor of the microprocessor miRNA-biogenetic inconsistency

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