Dataset Information

Auxin sensing is a property of an unstructured domain in the Auxin Response Factor ETTIN of Arabidopsis thaliana.

ABSTRACT: The plant hormone auxin regulates numerous aspects of the plant life cycle. Auxin signalling is mediated by auxin response factors (ARFs) that dimerise with modulating Aux/IAA repressors. ARF3 (ETTIN or ETT) is atypical as it does not interact with Aux/IAA repressors. It is proposed to be a non-canonical auxin sensor, regulating diverse functions essential for development. This sensing ability relies on a unique C-terminal ETT specific domain (ES domain). Alignments of ETT orthologues across the angiosperm phylum revealed that the length and sequence identities of ES domains are poorly conserved. Computational predictors suggested the ES domains to be intrinsically disordered, explaining their tolerance of insertions, deletions and mutations during evolution. Nevertheless, five highly conserved short linear motifs were identified suggesting functional significance. High-throughput library screening identified an almost full-length soluble ES domain that did not bind auxin directly, but exhibited a dose-dependent response in a yeast two-hybrid system against the Arabidopsis INDEHISCENT (IND) transcription factor. Circular dichroism confirmed the domain was disordered. The identification and purification of this domain opens the way to the future characterisation of the ETT auxin-sensing mechanism in planta and an improved understanding of auxin-mediated regulation.

SUBMITTER: Simonini S

PROVIDER: S-EPMC6131142 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature

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Auxin sensing is a property of an unstructured domain in the Auxin Response Factor ETTIN of Arabidopsis thaliana.

Simonini Sara S Mas Philippe J PJ Mas Caroline M V S CMVS Østergaard Lars L Hart Darren J DJ

Scientific reports 20180910 1

The plant hormone auxin regulates numerous aspects of the plant life cycle. Auxin signalling is mediated by auxin response factors (ARFs) that dimerise with modulating Aux/IAA repressors. ARF3 (ETTIN or ETT) is atypical as it does not interact with Aux/IAA repressors. It is proposed to be a non-canonical auxin sensor, regulating diverse functions essential for development. This sensing ability relies on a unique C-terminal ETT specific domain (ES domain). Alignments of ETT orthologues across the ...[more]

PMID: 30202032

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Project description:BackgroundAuxins play key roles in the phytohormone network. Early auxin response genes in the AUX/IAA, SAUR, and GH3 families show functional redundancy, which makes it very difficult to study the functions of individual genes based on gene knockout analysis or transgenic technology. As an alternative, chemical genetics provides a powerful approach that can be used to address questions relating to plant hormones.ResultsBy screening a small-molecule chemical library of compounds that can induce abnormal seedling and vein development, we identified and characterized a piperazine compound 1-[(4-bromophenoxy) acetyl]-4-[(4-fluorophenyl) sulfonyl] piperazine (ASP). The Arabidopsis DR5::GFP line was used to assess if the effects mentioned were correlated with the auxin response, and we accordingly verified that ASP altered the auxin-related pathway. Subsequently, we examined the regulatory roles of ASP in hypocotyl and root development, auxin distribution, and changes in gene expression. Following ASP treatment, we detected hypocotyl elongation concomitant with enhanced cell elongation. Furthermore, seedlings showed retarded primary root growth, reduced gravitropism and increased root hair development. These phenotypes were associated with an increased induction of DR5::GUS expression in the root/stem transition zone and root tips. Auxin-related mutants including tir1-1, aux1-7 and axr2-1 showed phenotypes with different root-development pattern from that of the wild type (Col-0), and were insensitive to ASP. Confocal images of propidium iodide (PI)-stained root tip cells showed no detectable damage by ASP. Furthermore, RT-qPCR analyses of two other genes, namely, Ethylene Response Factor (ERF115) and Mediator 18 (MED18), which are related to cell regeneration and damage, indicated that the ASP inhibitory effect on root growth was not attributable to toxicity. RT-qPCR analysis provided further evidence that ASP induced the expression of early auxin-response-related genes.ConclusionsASP altered the auxin response pathway and regulated Arabidopsis growth and development. These results provide a basis for dissecting specific molecular components involved in auxin-regulated developmental processes and offer new opportunities to discover novel molecular players involved in the auxin response.

Project description:Background and aimsLATERAL ORGAN BOUNDARIES DOMAIN 29 (LBD29), an important molecule downstream of auxin response factors ARF7 and ARF19, has a critical role in lateral root formation in Arabidopsis thaliana. The cell cycle activation of pericycle cells and their speciﬁcation triggered by auxin are crucial for the initiation of lateral roots. In this study, we attempted to determine whether LBD29 is involved in auxin signalling and/or cell cycle regulation and to characterize the roles of LBD29 in these processes.MethodsThe impact of LBD29 on cell cycling progression in pericycle cells was investigated in lbd29 loss-of-function mutant or LBD29-over-expressing plants. The cell cycle was determined by measuring the expression of some cell cycle-related genes using in situ hybridization and quantitative real-time reverse transcription-PCR (qRT-PCR). Furthermore, the cell division in the root explants from either the lbd29 mutant, LBD29-over-expressing plants or the wild type grown in auxin-rich media was also analysed and compared by the distribution of DR5:β-glucuronidase (GUS) in the primordia or by the expression of PIN-FORMED (PIN) members and PLETHROA 1 (PLT1) which represented the auxin response by the pericycle cells.Key resultslbd29 mutation resulted in reduced numbers of lateral roots and primordia, whereas LBD29 over-expression resulted in more lateral root and primordia formation than in the wild type. More importantly, the level of LBD29 expression was found to be positively correlated with the level of expression of cell cycle-related genes and correlated with the numbers of subcellular organelles found in pericycle cells in the maturation zone. In addition, an in vitro experiment using root explants demonstrated that the presence of LBD29 was required for the maintenance of the cell division capacity of the pericycle. Furthermore, LBD29 appeared to modify PIN-dependent auxin signalling in the primordia since there was a correlated association between the expression of PINs, PLT1 and DR5:GUS and the expression of LBD29.ConclusionsThe ability of LBD29 to regulate lateral root initiation is associated with its maintenance of the cell division capacity of the pericycle in response to auxin and its involvement in the auxin signalling pathway.

Dataset Information

Auxin sensing is a property of an unstructured domain in the Auxin Response Factor ETTIN of Arabidopsis thaliana.

Publications

Auxin sensing is a property of an unstructured domain in the Auxin Response Factor ETTIN of Arabidopsis thaliana.

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