biostudies-literatureMickleburgh IWellcome TrustBiotechnology and Biological Sciences Research Council8605-20https://www.ebi.ac.uk/biostudies/studies/S-EPMC4117754biostudies-literatureUnknown42(13)Post-transcriptional steps of gene expression are regulated by RNA binding proteins. Major progress has been made in characterizing RNA-protein interactions, from high resolution structures to transcriptome-wide profiling. Due to the inherent technical challenges, less attention has been paid to the way in which proteins with multiple RNA binding domains engage with target RNAs. We have investigated how the four RNA recognition motif (RRM) domains of Polypyrimidine tract binding (PTB) protein, a major splicing regulator, interact with FAS pre-mRNA under conditions in which PTB represses FAS exon 6 splicing. A combination of tethered hydroxyl radical probing, targeted inactivation of individual RRMs and single molecule analyses revealed an unequal division of labour between the four RRMs ofNucleic acids researchThe organization of RNA contacts by PTB for regulation of FAS splicing.PMC4117754092900BB/E02209X/1BB/H004203/1088113/Z/08/ZKafasla PLlorian MMickleburgh IJackson RJSmith CWCherny DCurry SfalseThe organization of RNA contacts by PTB for regulation of FAS splicing.Post-transcriptional steps of gene expression are regulated by RNA binding proteins. Major progress has been made in characterizing RNA-protein interactions, from high resolution structures to transcriptome-wide profiling. Due to the inherent technical challenges, less attention has been paid to the way in which proteins with multiple RNA binding domains engage with target RNAs. We have investigated how the four RNA recognition motif (RRM) domains of Polypyrimidine tract binding (PTB) protein, a major splicing regulator, interact with FAS pre-mRNA under conditions in which PTB represses FAS exon 6 splicing. A combination of tethered hydroxyl radical probing, targeted inactivation of individual RRMs and single molecule analyses revealed an unequal division of labour between the four RRMs of2014-01-01T00:00:00Z2014 Jul2025-04-04T10:08:19.223Z2019-03-27T01:33:09ZS-EPMC41177542495760210.1093/nar/gku519