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Updated: Feb 19, 2026
RNA Blot Analysis for the Detection and Quantification of Plant MicroRNAs
Published on: July 11, 2020
A Role for the F-Box Protein HAWAIIAN SKIRT in Plant microRNA Function
Patricia L M Lang1, Michael D Christie1, Ezgi S Dogan1
1Max Planck Institute for Developmental Biology, Tübingen, Germany.
Abstract:
As regulators of gene expression in multicellular organisms, microRNAs (miRNAs) are crucial for growth and development. Although a plethora of factors involved in their biogenesis and action in Arabidopsis (Arabidopsis thaliana) has been described, these processes and their fine-tuning are not fully understood. Here, we used plants expressing an artificial miRNA target mimic (MIM) to screen for negative regulators of miR156. We identified a new mutant allele of the F-box gene HAWAIIAN SKIRT (HWS; At3G61590), hws-5, as a suppressor of the MIM156-induced developmental and molecular phenotypes. In hws plants, levels of some endogenous miRNAs are increased and their mRNA targets decreased. Plants constitutively expressing full-length HWS-but not a truncated version lacking the F-box domain-display morphological and molecular phenotypes resembling those of mutants defective in miRNA biogenesis and activity. In combination with such mutants, hws loses its delayed floral organ abscission ("skirt") phenotype, suggesting epistasis. Also, the hws transcriptome profile partially resembles those of well-known miRNA mutants hyl1-2, se-3, and ago1-27, pointing to a role in a common pathway. We thus propose HWS as a novel, F-box dependent factor involved in miRNA function.
Insights
Researchers identified the HAWAIIAN SKIRT (HWS) gene as a novel F-box protein regulating microRNA (miRNA) function. Mutations in HWS increase miRNA levels and affect plant development, suggesting a role in miRNA pathways.
Area of Science:
- Plant Molecular Biology
- Gene Regulation
- Developmental Biology
Background:
- MicroRNAs (miRNAs) are key regulators of gene expression in multicellular organisms, essential for growth and development.
- While many factors influencing miRNA biogenesis and function in *Arabidopsis thaliana* are known, their complete understanding and fine-tuning remain incomplete.
Purpose of the Study:
- To screen for negative regulators of miR156 using an artificial miRNA target mimic (MIM) system.
- To investigate the role of the F-box gene HAWAIIAN SKIRT (HWS) in miRNA regulation and plant development.
Main Methods:
- Utilized an artificial miRNA target mimic (MIM156) screen in *Arabidopsis thaliana*.
- Characterized a new mutant allele, *hws*-*5*, and analyzed plants expressing full-length and truncated HWS.
- Performed transcriptome profiling and genetic analysis (epistasis studies) with known miRNA pathway mutants.
Main Results:
- Identified *hws*-*5* as a suppressor of MIM156-induced phenotypes.
- Observed increased endogenous miRNA levels and decreased mRNA targets in *hws* mutants.
- Demonstrated that constitutive HWS expression mimics miRNA biogenesis/activity mutant phenotypes and that HWS acts in a common pathway with known miRNA regulators.
Conclusions:
- Propose HAWAIIAN SKIRT (HWS) as a novel, F-box dependent factor involved in microRNA (miRNA) function.
- HWS plays a significant role in the regulation of miRNA levels and/or activity, impacting plant development.
- The findings suggest HWS acts within a conserved pathway critical for miRNA-mediated gene regulation.
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