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Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip
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Genome-wide mRNA surveillance is coupled to mRNA export

Haley Hieronymus1, Michael C Yu, Pamela A Silver

  • 1Department of Systems Biology, Harvard Medical School and the Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA.

Genes & Development
|October 19, 2004
PubMed

Insights

Researchers discovered new factors linking mRNA export to DNA and RNA surveillance. The study reveals how DNA damage affects mRNA export and degradation, involving specific proteins like Rrp6 and Lrp1.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Nuclear export of messenger RNA (mRNA) is crucial for gene expression, tightly linked with transcription and RNA processing.
  • The impact of DNA damage on mRNA fate and export remains largely unexplored.
  • Understanding these processes is vital for comprehending cellular responses to stress and maintaining genomic integrity.

Purpose of the Study:

  • To identify novel factors involved in mRNA export using a comprehensive genetic screen in Saccharomyces cerevisiae.
  • To investigate the interplay between DNA damage, RNA surveillance, and mRNA export pathways.
  • To elucidate the roles of newly identified factors, Rrp6 and Lrp1, in these cellular processes.

Main Methods:

  • Conducted a genome-wide screen of nonessential Saccharomyces cerevisiae genes to identify factors affecting mRNA export.
  • Utilized co-immunoprecipitation to assess physical interactions between identified factors.
  • Performed gene localization studies to determine genomic association with target genes.
  • Assessed the role of identified factors in DNA repair following UV irradiation.

Main Results:

  • Discovered four new factors essential for mRNA export, including mRNA surveillance factor Rrp6 and DNA repair protein Lrp1.
  • Established that Rrp6 and Lrp1 are nuclear exosome components that physically interact.
  • Demonstrated that Lrp1 mediates both specific and general mRNA degradation, particularly upon UV-induced DNA damage.
  • Showed that Lrp1 requires Rrp6 for genomic localization to target genes, and Rrp6 localization correlates with transcription.
  • Found that both Rrp6 and Lrp1 are essential for repairing UV-induced DNA damage.

Conclusions:

  • The study reveals a significant coupling between mRNA surveillance mechanisms and mRNA export pathways.
  • Findings suggest that the RNA surveillance machinery exhibits specificity towards different transcript populations.
  • This research establishes a direct link between DNA surveillance, RNA surveillance, and the process of mRNA export.

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