Functional analysis of molecular interactions in synthetic auxin response circuits

Auxin-regulated transcription pivots on the interaction between the AUXIN/INDOLE-3-ACETIC ACID (Aux/IAA) repressor proteins and the AUXIN RESPONSE FACTOR (ARF) transcription factors. Recent structural analyses of ARFs and Aux/IAAs have raised questions about the functional complexes driving auxin tr...

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Published in	Proceedings of the National Academy of Sciences - PNAS Vol. 113; no. 40; pp. 11354 - 11359
Main Authors	Pierre-Jerome, Edith, Moss, Britney L., Lanctot, Amy, Hageman, Amber, Nemhauser, Jennifer L.
Format	Journal Article
Language	English
Published	United States National Academy of Sciences 04.10.2016
Subjects	Acetic acid Arabidopsis - drug effects Arabidopsis - genetics Biological Sciences Deoxyribonucleic acid DNA Flowers & plants Gene Expression Regulation, Plant - drug effects Gene Regulatory Networks Indoleacetic Acids - pharmacology Models, Biological Mutation - genetics Plants, Genetically Modified Promoter Regions, Genetic - genetics Protein Multimerization Proteins Recombinant Fusion Proteins - metabolism Response Elements - genetics Transcription, Genetic - drug effects Transgenes Yeast Yeasts synthetic circuits transcription ARF auxin signaling PB1
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ISSN	0027-8424 1091-6490 1091-6490
DOI	10.1073/pnas.1604379113

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Summary:	Auxin-regulated transcription pivots on the interaction between the AUXIN/INDOLE-3-ACETIC ACID (Aux/IAA) repressor proteins and the AUXIN RESPONSE FACTOR (ARF) transcription factors. Recent structural analyses of ARFs and Aux/IAAs have raised questions about the functional complexes driving auxin transcriptional responses. To parse the nature and significance of ARF–DNA and ARF–Aux/IAA interactions, we analyzed structure-guided variants of synthetic auxin response circuits in the budding yeast Saccharomyces cerevisiae. Our analysis revealed that promoter architecture could specify ARF activity and that ARF19 required dimerization at two distinct domains for full transcriptional activation. In addition, monomeric Aux/IAAs were able to repress ARF activity in both yeast and plants. This systematic, quantitative structure-function analysis identified a minimal complex—comprising a single Aux/IAA repressing a pair of dimerized ARFs—sufficient for auxin-induced transcription.
Bibliography:	SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 14 ObjectType-Article-1 ObjectType-Feature-2 content type line 23 Edited by Athanasios Theologis, Plant Gene Expression Center, Albany, CA, and approved July 29, 2016 (received for review March 16, 2016) Author contributions: E.P.-J., B.L.M., and J.L.N. designed research; E.P.-J., B.L.M., and A.L. performed research; A.H. contributed new reagents/analytic tools; E.P.-J., B.L.M., and A.L. analyzed data; and E.P.-J. and J.L.N. wrote the paper.
ISSN:	0027-8424 1091-6490 1091-6490
DOI:	10.1073/pnas.1604379113