Deciphering the Dynamic Landscape of Transcription-Associated mRNP Quality Control Components Over the Whole Yeast Genome

Methods Mol Biol. 2021:2209:251-265. doi: 10.1007/978-1-0716-0935-4_16.

Abstract

In eukaryotic cells, aberrant mRNPs with processing and packaging defects are targeted co-transcriptionally by a surveillance system that triggers their nuclear retention and ultimately the degradation of their mRNA component by the 3'-5' activity of the exosome-associated exonuclease Rrp6. This mRNP quality control process is stimulated by the NNS complex (Nrd1-Nab3-Sen1), which otherwise mediates termination, processing, and decay of ncRNAs. The process involves also the exosome co-activator TRAMP complex (Trf4-Air2-Mtr4). Here, we describe a genome-wide approach to visualize the dynamic movement and coordination of these quality control components over the yeast chromosomes upon perturbation of mRNP biogenesis. The method provides valuable information on how the surveillance system is precisely coordinated both physically and functionally with the transcription machinery to detect the faulty events during perturbation of mRNP biogenesis. The overview shows also that the gathering of the quality control components over affected mRNA genes takes place at the expense of their commitment to be recruited at ncRNA genomic features, provoking termination and processing defects of ncRNAs.

Keywords: ChIP-seq; RNA-seq; Yeast; mRNP biogenesis; mRNP quality control; ncRNAs.

MeSH terms

  • Gene Expression Regulation, Fungal
  • High-Throughput Nucleotide Sequencing / methods
  • RNA, Fungal / genetics*
  • RNA, Messenger / genetics*
  • RNA, Untranslated / genetics*
  • RNA-Binding Proteins / chemistry*
  • Ribonucleoproteins / genetics*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / chemistry*
  • Transcription, Genetic

Substances

  • RNA, Fungal
  • RNA, Messenger
  • RNA, Untranslated
  • RNA-Binding Proteins
  • Ribonucleoproteins
  • Saccharomyces cerevisiae Proteins
  • messenger ribonucleoprotein