The CUP-SHAPED COTYLEDON 2 - MIR164A regulatory feedback loop directs leaf morphogenesis
- 1INRAE, Institut Jean-Pierre Bourgin (IJPB), INRAE-AgroParisTech INRA Centre de Versailles-Grignon
Abstract
Gene regulatory networks (GRNs) establish the spatio-temporal gene expression patterns necessary for reproducible development of organisms. MicroRNAs have emerged as important regulators of gene expression controlling either target gene levels, spatial patterns or timing. These molecular functions may vary depending on how they are integrated into GRNs. Here we used the Arabidopsis leaf as a model to study the genetic and molecular interactions between the CUP-SHAPED COTYLEDON 2 (CUC2) transcription factor and its negative regulator, miR164A. Quantitative morphometric analyses showed that miR164A acts from early stages during leaf development regulating CUC2 expression dynamic rather than the spatial distribution of CUC2. At the molecular level, CUC2 directly activates MIR164A expression, forming a single negative feedback loop. Further, using a ratiometric approach, we identified a cis-regulatory element (CRE) sufficient to drive CUC2-dependent MIR164A expression at the leaf sinus. Ordinary differential equation (ODE) modelling suggests that CUC2-CRE may control the timely accumulation of CUC2. Accordingly, quantification of CUC2 protein accumulation dynamics using a spatio-temporal mean cartography strategy in CRISPR cis-regulatory variants at the MIR164A locus revealed that CUC2-dependent MIRA164A expression is required to maintain both the precise timing of CUC2 expression and CUC2 levels. Altogether, our work provides experimental evidence for a specific role of the CUC2-MIR164A module in shaping leaf development, highlighting the contribution of network topology to GRN functions.
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BibTeX
@misc{bigot2026cupshaped,
title = {{The CUP-SHAPED COTYLEDON 2 - MIR164A regulatory feedback loop directs leaf morphogenesis}},
author = {S. Bigot and A. Maugarny and N. Verdon and A. Vialette and M. Goussot and F. Roudier and J. Burguet and P. Laufs and N. Arnaud},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.07.757447},
url = {https://www.biorxiv.org/content/10.64898/2026.10.07.757447v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - The CUP-SHAPED COTYLEDON 2 - MIR164A regulatory feedback loop directs leaf morphogenesis
AU - S. Bigot
AU - A. Maugarny
AU - N. Verdon
AU - A. Vialette
AU - M. Goussot
AU - F. Roudier
AU - J. Burguet
AU - P. Laufs
AU - N. Arnaud
PY - 2026
DA - 2026/10/07
PB - bioRxiv
DO - 10.64898/2026.10.07.757447
UR - https://www.biorxiv.org/content/10.64898/2026.10.07.757447v1
AB - Gene regulatory networks (GRNs) establish the spatio-temporal gene expression patterns necessary for reproducible development of organisms. MicroRNAs have emerged as important regulators of gene expression controlling either target gene levels, spatial patterns or timing. These molecular functions may vary depending on how they are integrated into GRNs. Here we used the Arabidopsis leaf as a model to study the genetic and molecular interactions between the CUP-SHAPED COTYLEDON 2 (CUC2) transcription factor and its negative regulator, miR164A. Quantitative morphometric analyses showed that miR164A acts from early stages during leaf development regulating CUC2 expression dynamic rather than the spatial distribution of CUC2. At the molecular level, CUC2 directly activates MIR164A expression, forming a single negative feedback loop. Further, using a ratiometric approach, we identified a cis-regulatory element (CRE) sufficient to drive CUC2-dependent MIR164A expression at the leaf sinus. Ordinary differential equation (ODE) modelling suggests that CUC2-CRE may control the timely accumulation of CUC2. Accordingly, quantification of CUC2 protein accumulation dynamics using a spatio-temporal mean cartography strategy in CRISPR cis-regulatory variants at the MIR164A locus revealed that CUC2-dependent MIRA164A expression is required to maintain both the precise timing of CUC2 expression and CUC2 levels. Altogether, our work provides experimental evidence for a specific role of the CUC2-MIR164A module in shaping leaf development, highlighting the contribution of network topology to GRN functions.
ER -