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Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo
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Class I TCP transcription factor AtTCP8 modulates key brassinosteroid-responsive genes
Benjamin J Spears1,2,3, Samuel A McInturf2,3, Carina Collins4
1Department of Biological Sciences, Butler University, Indianapolis, Indiana, USA.
Plant Physiology
|July 22, 2022
Summary
The TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) transcription factor TCP8 regulates plant development and immunity by modulating brassinosteroid signaling. Pathogens may target TCP8 to disrupt these crucial plant pathways.
Area of Science:
- Plant molecular biology
- Plant signaling pathways
- Transcription factors
Background:
- TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) transcription factors regulate plant development and immunity.
- TCPs are targeted by pathogen virulence factors and regulate plant defense genes, but their targets are not fully characterized.
- Loss of function in AtTCP8, AtTCP14, and AtTCP15 affects Arabidopsis thaliana immune signaling.
Purpose of the Study:
- To comprehensively characterize the gene targets of the plant-specific TCP transcription factor, AtTCP8.
- To investigate the role of AtTCP8 in plant hormone signaling pathways, particularly brassinosteroids (BRs).
- To understand how AtTCP8's function relates to its targeting by pathogen effectors.
Main Methods:
- Chromatin immunoprecipitation sequencing (ChIP-seq) to identify AtTCP8-bound gene promoters.
- RNA sequencing (RNA-seq) to identify differentially regulated genes in tcp8 mutants.
- Analysis of phytohormone signaling components, focusing on BR signaling genes like BZR1 and BZR2/BES1.
Main Results:
- TCP8-bound promoters and differentially regulated genes were enriched in signaling components of BR, auxin, and jasmonic acid pathways.
- AtTCP8 directly binds and activates the promoters of key BR transcriptional regulators, BZR1 and BZR2/BES1.
- tcp8 mutant seedlings showed altered BR-responsive growth, reduced BZR2 transcript levels, and BR-responsive changes in TCP8 localization and activity.
Conclusions:
- AtTCP8 acts as a direct modulator of brassinosteroid signaling by regulating BZR1 and BZR2/BES1 expression.
- TCP8's role in modulating plant hormone pathways, including BR signaling, provides a potential explanation for its targeting by pathogen effectors.
- This study elucidates a key mechanism by which TCP transcription factors integrate developmental and immune signaling in plants.
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