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Widespread Polycistronic Transcripts in Fungi Revealed by Single-Molecule mRNA Sequencing.

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Area of Science:

  • Fungal genomics
  • Transcriptomics
  • Molecular biology

Background:

  • Genes in prokaryotes are often organized into polycistronic transcripts.
  • Polycistronic transcription in eukaryotes was considered rare, with limited evidence in higher eukaryotes.
  • Understanding fungal gene regulation is crucial for mycology and biotechnology.

Purpose of the Study:

  • To investigate the prevalence and characteristics of polycistronic transcription in mushroom-forming fungi (Agaricomycetes).
  • To develop and validate a long-read sequencing strategy for identifying complex fungal transcriptomes.
  • To explore the functional implications and evolutionary conservation of fungal polycistronic transcription.

Main Methods:

  • Employed a long-read sequencing strategy to capture full-length transcripts.
  • Developed a bioinformatics pipeline for analyzing complex transcriptomes and identifying polycistronic RNAs.
  • Conducted comparative genomic analysis across multiple fungal species.

Main Results:

  • Discovered genome-wide prevalence of polycistronic transcription in Agaricomycetes, involving up to 8% of transcribed genes.
  • Demonstrated that fungal polycistronic transcripts differ from prokaryotic ones, with co-transcribed genes also being independently transcribed.
  • Showed that polycistronic transcription can potentially interfere with the expression of downstream genes.
  • Identified polycistronic transcription as a conserved feature across a wide range of mushroom-forming fungi.

Conclusions:

  • Polycistronic transcription is a widespread phenomenon in higher fungi, not a rare occurrence.
  • The developed long-read sequencing and bioinformatics approach is a powerful tool for characterizing complex transcriptomes, including novel mRNA isoforms.
  • This study significantly advances our understanding of fungal gene expression and transcriptome complexity.