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RNA G-quadruplexes regulate mammalian mirtron biogenesis.

Uzma Salim1, Manoj B Menon1, Sonam Dhamija2

  • 1Kusuma School of Biological Sciences, Indian Institute of Technology, New Delhi, India.

The Journal of Biological Chemistry
|February 8, 2025
PubMed
Summary

Mammalian mirtrons utilize RNA G-quadruplexes (rG4s) in their 5' arm to facilitate splicing-dependent biogenesis. Disrupting these rG4s inhibits mirtron maturation, revealing their crucial regulatory role.

Keywords:
Drosha-independent processingG-quadruplexRNA G-quadruplex(rG4)miRNAmirtronsnoncanonical miRNAsplicing

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Area of Science:

  • Molecular Biology
  • RNA Biology
  • Genetics

Background:

  • Mirtrons are noncanonical microRNAs processed via splicing, independent of Drosha.
  • The precise mechanisms governing mirtron biogenesis are not fully understood.
  • Potential interference of mirtron splicing with canonical microRNA pathways exists.

Purpose of the Study:

  • To elucidate the regulatory mechanisms of mirtron biogenesis.
  • To investigate sequence and structural features differentiating plant, invertebrate, and vertebrate mirtrons.
  • To determine the role of RNA G-quadruplexes (rG4s) in mammalian mirtron formation.

Main Methods:

  • Comparative sequence analysis of mirtrons across different species.
  • In silico prediction and in vitro structural analysis of RNA G-quadruplexes (rG4s).
  • Mutagenesis studies to assess the impact of rG4 disruption on splicing and maturation.

Main Results:

  • Vertebrate, particularly mammalian, mirtrons show a distinct enrichment of guanines in their 5' arm compared to uracil-enriched plant/invertebrate mirtrons.
  • Most mammalian mirtrons contain RNA G-quadruplexes (rG4s), predominantly located in the 5' arm.
  • In vitro experiments confirmed rG4 formation in human mirtrons, and disruption of these rG4s significantly inhibited splicing and maturation, while other mutations had no effect.

Conclusions:

  • RNA G-quadruplexes (rG4s) in the 5' arm are critical regulatory elements for mammalian mirtron biogenesis.
  • rG4 formation facilitates the splicing-dependent maturation pathway of mammalian mirtrons.
  • This study highlights a novel role for rG4s in small RNA biology and mirtron evolution.