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The ThbZIP1 can be regulated by an ABF Transcription Factor and sensitivity to ABA treatment

ABSTRACT: In the current report, we report that ThbZIP1 is a direct target gene of the ThABF1 transcription factor. There are three ABRE motifs in the promoter of ThbZIP1, Yeast one-hybrid (Y1H) assays showed that a ABF protein, ThABF1, specifically binds to the ABRE motifs. The interaction between ThABF1 and the promoter of ThbZIP1 was further confirmed by transient expression assays in tobacco leaves. Chromatin Immunoprecipitation (ChIP) results suggested that binding of ThABF1 to ABRE motifs in the promoter of ThbZIP1 occurs in vivo in Tamarix hispida to regulate the expression of ThbZIP1. Moreover, ThABF1 and ThbZIP1 share similar expression patterns in response to salt, drought, ABA, methyl viologen (MV) and cold stress. Microarray analyses results showed there were 1,662 and 1,609 genes that were significantly upregulated or downregulated, respectively, under ABA stress conditions. ThbZIP1 regulated the genes via binding to the C-, G- or A-box motifs in their promoter sequences. Based on these data, the results suggested a regulatory network model mediated by ThbZIP1, under abiotic stress conditions, ThABF1 regulates the expression of ThbZIP1, and the activated ThbZIP1 binds to bZIP recognition sequences or other motifs to regulate the expression of genes containing these motifs in their promoters.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE62888 | GEO | 2014/11/01

SECONDARY ACCESSION(S): PRJNA265947

REPOSITORIES: GEO

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Project description:The aim of this experiment was to decipher a transcriptional regulatory code of Arabidopsis Auxin Response Factor 2 (ARF2) and understand the molecular basis of ARF2’s role as a promoter of dark-induced senescence. A combination of computational tools and yeast one-hybrid experiments identified potential direct transcriptional regulators of ARF2 belonging to several TF families including AP2/ERFs and AREB/ABF/ABI5s. Using ARF2 promoter mutants in transcriptional activation and Y1H experiments, the binding sites of these TFs in the ARF2 promoter were identified. AP2/ERFs including ERF1, ERF15 and ORA59 transactivate the ARF2 promoter in Arabidopsis mesophyll protoplasts and this transactivation is positively regulated by ABA. Crucially, the positive effect of ABA on ERF1 transactivation of ARF2 relies on the functional ABRE motif in the ARF2 promoter and the presence of AREB/ABF/ABI5s (ABF3 and ABF4) suggesting the existence of combinatorial regulation of ARF2 by these direct regulators. ABF3 and ABF4 were unable to activate the ARF2 promoter on their own indicating the necessity of ERF1 as a coupling TF for AREB/ABF/ABI5-driven regulation of ARF2. We also show that MED25 (a subunit of the Mediator complex) is required for proper transcriptional regulation of ARF2 during senescence. med25 plants show decreased ARF2 mRNA levels during natural senescence and, similarly to arf2-6 mutants, exhibit a delayed dark-induced senescence phenotype. We show that MED25 is important for ERF1-, ERF15- and ORA59-dependent transactivation of ARF2 in Arabidopsis protoplasts and finally idenitfy genes differentially expressed in arf2-6 mutants compared to wildtype during dark-induced senescence. Hence, we elucidate a regulatory network around ARF2 identifying combinatorial transcriptional regulation of this gene during senescence as well as likely downstream targets. The combinatorial regulation, comprised of AP2/ERFs and AREB/ABF/ABI5s TFs, enables integration of at least two key hormone signaling pathways to drive expression of a key senescence regulator.

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The ThbZIP1 can be regulated by an ABF Transcription Factor and sensitivity to ABA treatment

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