Related Experiment Video

Updated: Sep 13, 2025

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
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Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro

Published on: May 3, 2014

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Pre-mRNA processing factors differentially impact coordination between co-transcriptional cleavage and transcription

Xianhao Jin1, Juzuo Li1, Wenqin Lu1

  • 1Shenzhen Key Laboratory of Plant Genetic Engineering and Molecular Design, Institute of Plant and Food Science, Department of Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, China.

Nature Communications
|August 1, 2025
PubMed

Insights

mRNA processing factors coordinate co-transcriptional cleavage and transcription termination. New single-molecule sequencing reveals diverse roles, showing how these events are tightly linked during gene expression in plants.

Area of Science:

  • Molecular Biology
  • Plant Science
  • RNA Biology

Background:

  • Co-transcriptional cleavage and transcription termination are crucial for mRNA maturation but their coordination is poorly understood.
  • Detecting these transient molecular events in real-time has been a significant challenge in RNA biology.

Purpose of the Study:

  • To investigate the coordination between co-transcriptional cleavage and transcription termination.
  • To elucidate the roles of specific pre-mRNA processing factors in linking these two essential mRNA maturation events.

Main Methods:

  • Application of single-molecule nascent RNA sequencing to simultaneously assess cleavage status and readthrough distance on individual RNA molecules.
  • Characterization of 14 pre-mRNA processing factor mutants in Arabidopsis to analyze their impact on cleavage and termination.

Main Results:

  • Core CPSF and CstF complex components promote both cleavage and termination, aiding exoribonuclease AtXRN3 access.
  • Mutations in PAPS1 and AtXRN3 delayed termination, while BORDER proteins inhibited cleavage and facilitated termination.
  • SSU72 specifically enhanced termination, and FPA promoted termination of cleaved readthrough transcripts.

Conclusions:

  • Pre-mRNA processing factors play diverse and coordinated roles in linking co-transcriptional cleavage and transcription termination.
  • The study provides a comprehensive dataset revealing the intricate coordination between cleavage and termination during pre-mRNA processing in plants.

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