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Unpaired nucleotides on the stem of microRNA precursor are important for precise cleavage by Dicer-like 1 in

Rikako Hirata1, Tomoya Makabe1, Kei-Ichiro Mishiba1

  • 1Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Japan.

Genes to Cells : Devoted to Molecular & Cellular Mechanisms
|February 10, 2022
PubMed

Insights

Mismatches on plant primary microRNA (pri-miRNA) stems are crucial for precise Dicer-like 1 (DCL1) processing. These mismatches guide DCL1 activity, ensuring correct microRNA generation.

Area of Science:

  • Plant molecular biology
  • RNA biology
  • Biochemistry

Background:

  • Dicer-like 1 (DCL1) is essential for plant microRNA (miRNA) biogenesis.
  • Primary miRNA (pri-miRNA) is processed into miRNA/miRNA* duplexes by DCL1.
  • Pri-miRNA features a stem-loop structure with mismatches on the stem.

Purpose of the Study:

  • To investigate the role of mismatches on pri-miRNA stems in DCL1 cleavage.
  • To determine how mismatches influence the precision of DCL1 processing.

Main Methods:

  • In vitro cleavage assays using purified recombinant Arabidopsis DCL1.
  • Analysis of pri-miRNA variants with specific mismatch mutations.

Main Results:

  • Mismatches near the miRNA/miRNA* duplex are important for efficient pri-miRNA cleavage.
  • Mismatches distant from the duplex determine DCL1 cleavage position.
  • Mutant pri-miRNAs lacking specific mismatches were cleaved at aberrant positions, producing non-miRNA small RNAs.

Conclusions:

  • Stem mismatches on pri-miRNA are key determinants for precise DCL1 processing in plants.
  • DCL1 cleavage precision is influenced by both distance from the ssRNA-dsRNA junction and stem mismatches.
Keywords:
Arabidopsis thalianaDicerRNA secondary structuremicroRNAmicroRNA biogenesis

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