Identifying eIF4E-binding protein translationally-controlled transcripts reveals links to mRNAs bound by specific PUF proteins

Nucleic Acids Res. 2010 Dec;38(22):8039-50. doi: 10.1093/nar/gkq686. Epub 2010 Aug 12.

Abstract

eIF4E-binding proteins (4E-BPs) regulate translation of mRNAs in eukaryotes. However the extent to which specific mRNA targets are regulated by 4E-BPs remains unknown. We performed translational profiling by microarray analysis of polysome and monosome associated mRNAs in wild-type and mutant cells to identify mRNAs in yeast regulated by the 4E-BPs Caf20p and Eap1p; the first-global comparison of 4E-BP target mRNAs. We find that yeast 4E-BPs modulate the translation of >1000 genes. Most target mRNAs differ between the 4E-BPs revealing mRNA specificity for translational control by each 4E-BP. This is supported by observations that eap1Δ and caf20Δ cells have different nitrogen source utilization defects, implying different mRNA targets. To account for the mRNA specificity shown by each 4E-BP, we found correlations between our data sets and previously determined targets of yeast mRNA-binding proteins. We used affinity chromatography experiments to uncover specific RNA-stabilized complexes formed between Caf20p and Puf4p/Puf5p and between Eap1p and Puf1p/Puf2p. Thus the combined action of each 4E-BP with specific 3'-UTR-binding proteins mediates mRNA-specific translational control in yeast, showing that this form of translational control is more widely employed than previously thought.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / metabolism
  • Cyclins / genetics
  • Eukaryotic Initiation Factors / genetics
  • Eukaryotic Initiation Factors / physiology*
  • Gene Deletion
  • Gene Expression Regulation, Fungal*
  • Nitrogen / metabolism
  • Oligonucleotide Array Sequence Analysis
  • Protein Biosynthesis*
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / metabolism*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / physiology*
  • Transcription Factors / genetics
  • Transcription Factors / physiology*
  • Transcription, Genetic

Substances

  • Amino Acids
  • Caf20 protein, S cerevisiae
  • Cyclins
  • Eap1 protein, S cerevisiae
  • Eukaryotic Initiation Factors
  • RNA, Messenger
  • RNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Nitrogen