pervasive and dynamic protein binding sites of the mrna transcriptome in saccharomyces cerevisiae普遍的和动态的蛋白质结合位点的mrna转录组在酿酒酵母.pdfVIP

Freeberg et al. Genome Biology 2013, 14:R13 /content/14/2/R13 RESEARCH Open Access Pervasive and dynamic protein binding sites of the mRNA transcriptome in Saccharomyces cerevisiae 1,2† 1,3† 4 1,2 5 5 Mallory A Freeberg , Ting Han , James J Moresco , Andy Kong , Yu-Cheng Yang , Zhi John Lu , John R Yates and John K Kim 1* Abstract Background: Protein-RNA interactions are integral components of nearly every aspect of biology, including regulation of gene expression, assembly of cellular architectures, and pathogenesis of human diseases. However, studies in the past few decades have only uncovered a small fraction of the vast landscape of the protein-RNA interactome in any organism, and even less is known about the dynamics of protein-RNA interactions under changing developmental and environmental conditions. Results: Here, we describe the gPAR-CLIP (global photoactivatable-ribonucleoside-enhanced crosslinking and immunopurification) approach for capturing regions of the untranslated, polyadenylated transcriptome bound by RNA-binding proteins (RBPs) in budding yeast. We report over 13,000 RBP crosslinking sites in untranslated regions (UTRs) covering 72% of protein-coding transcripts encoded in the genome, confirming 3’ UTRs as major sites for RBP interaction. Comparative genomic analyses reveal that RBP crosslinking sites are highly conserved, and RNA folding predictions indicate that secondary structural elements are constrained by protein binding and may serve as generalizable modes of RNA recognition. Finally, 38% of 3’ UTR crosslinking sites show changes in RBP occupancy upon glucose or nitrogen deprivation, with major impacts on metabolic pathways as well as mitochondrial and