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The spliceosome-associated protein CWC15 promotes miRNA biogenesis in Arabidopsis

Bangjun Zhou1,2, Huihui Yu1,2, Yong Xue1,2

  • 1Center for Plant Science Innovation, University of Nebraska-Lincoln, Lincoln, NE, 68588-0666, USA.

Nature Communications
|March 17, 2024
PubMed

Insights

CWC15, a splicing protein, enhances microRNA (miRNA) biogenesis by regulating transcription and processing. It maintains optimal miRNA production by controlling key protein levels and pri-miRNA transcription.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) are crucial regulators of gene expression.
  • miRNA biogenesis is tightly controlled by the microprocessor complex.
  • Understanding factors influencing miRNA production is vital for cellular regulation.

Purpose of the Study:

  • To investigate the role of CWC15, a spliceosome-associated protein, in miRNA biogenesis.
  • To elucidate the mechanisms by which CWC15 affects miRNA production.
  • To identify novel regulators of miRNA biogenesis.

Main Methods:

  • Chromatin immunoprecipitation (ChIP) to assess DNA-dependent RNA polymerase occupancy.
  • Co-immunoprecipitation (Co-IP) to study protein interactions.
  • Proteasome activity assays and Western blotting to analyze protein levels and phosphorylation.

Main Results:

  • CWC15 positively regulates the transcription of primary miRNA transcripts (pri-miRNAs) by binding to miRNA gene promoters.
  • CWC15 interacts with microprocessor components Serrate (SE) and HYL1, promoting pri-miRNA processing.
  • CWC15 facilitates SE phosphorylation and degradation, ensuring proper levels of microprocessor components.

Conclusions:

  • CWC15 is a positive regulator of miRNA biogenesis, acting at both transcriptional and post-transcriptional levels.
  • The study reveals a novel function for a conserved splicing-related protein in miRNA production.
  • CWC15 integrates splicing machinery with miRNA biogenesis pathways.

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