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Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing
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ARGONAUTE1-binding Tudor domain proteins function in small interfering RNA production for RNA-directed DNA
Takahito Takei1,2, Michio Tsukada3, Kentaro Tamura4
1Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan.
Plant Physiology
|March 6, 2024
Summary
Angiosperm-specific proteins PDS5C/E interact with AGO1 to regulate transposable elements (TEs) via 24-nt siRNA production, crucial for plant evolution and adaptation.
Area of Science:
- Plant molecular biology
- Epigenetics
- Genomics
Background:
- Transposable elements (TEs) drive plant evolution and adaptation, but their regulation is poorly understood.
- RNA-directed DNA methylation (RdDM) is a key mechanism for controlling TEs in plants.
- The cohesin complex, including PDS5 proteins, plays roles in genome stability.
Purpose of the Study:
- To investigate the function of angiosperm-specific PDS5C/E proteins in TE regulation.
- To identify novel interactors and mechanisms involved in RdDM.
- To understand the unique role of PDS5C/E in angiosperm development and adaptation.
Main Methods:
- Proteomics and co-immunoprecipitation to identify protein interactions.
- RNA sequencing to analyze transcriptomic changes in mutant lines.
- Small RNA sequencing to quantify siRNA abundance.
- DNA methylation analysis (CHH) to assess RdDM establishment.
Main Results:
- Novel ARGONAUTE 1 (AGO1)-binding proteins, PDS5C/E, were identified.
- The pds5c/e double mutant showed defects in 24-nt small interfering RNA (siRNA) accumulation and TE DNA methylation.
- PDS5C/E and AGO1 cooperate in the cytoplasm for 24-nt siRNA production.
- lncRNAs accumulating in pds5c/e mutants were targeted by 21-nt miRNAs and siRNAs.
Conclusions:
- Angiosperm-specific PDS5C/E proteins are novel regulators of TE control through RdDM.
- PDS5C/E and AGO1 function in the cytoplasm to promote 24-nt siRNA biogenesis.
- This pathway represents an evolved mechanism in angiosperms to manage TE proliferation during evolution.
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