title: Identifying eIF4E-binding protein translationally-controlled transcripts reveals links to mRNAs bound by specific PUF proteins.
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.
authors: The Michael Smith Building, Faculty of Life Sciences, University of Manchester, Manchester, UK.
authors: Andrew G
authors: AG
authors: Cridge
authors: Lydia M
authors: LM
authors: Castelli
authors: Julia B
authors: JB
authors: Smirnova
authors: Julian N
authors: JN
authors: Selley
authors: William
authors: W
authors: Rowe
authors: Simon J
authors: SJ
authors: Hubbard
authors: John E G
authors: JE
authors: McCarthy
authors: Mark P
authors: MP
authors: Ashe
authors: Christopher M
authors: CM
authors: Grant
authors: Graham D
authors: GD
authors: Pavitt
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D016213:Cyclins / Q000235:genetics; D039642:Eukaryotic Initiation Factors / Q000235:genetics / Q000502:physiology*; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D009584:Nitrogen / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic
chemical_list: D000596:Amino Acids; C555411:Caf20 protein, S cerevisiae; D016213:Cyclins; C555412:Eap1 protein, S cerevisiae; D039642:Eukaryotic Initiation Factors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D009584:Nitrogen