Schneider, TimTimSchneiderHung, Lee-HsuehLee-HsuehHungAziz, MasoodMasoodAzizWilmen, AnnaAnnaWilmenThaum, StephanieStephanieThaumWagner, JaquelineJaquelineWagnerJanowski, RobertRobertJanowskiMüller, SimonSimonMüllerSchreiner, SilkeSilkeSchreinerFriedhoff, PeterPeterFriedhoffHüttelmaier, StefanStefanHüttelmaierNiessing, DierkDierkNiessingSattler, MichaelMichaelSattlerSchlundt, AndreasAndreasSchlundtBindereif, AlbrechtAlbrechtBindereif2022-11-182019-10-222022-11-182019http://nbn-resolving.de/urn:nbn:de:hebis:26-opus-148931https://jlupub.ub.uni-giessen.de/handle/jlupub/9501http://dx.doi.org/10.22029/jlupub-8889How multidomain RNA-binding proteins recognize their specific target sequences, based on a combinatorial code, represents a fundamental unsolved question and has not been studied systematically so far. Here we focus on a prototypical multidomain RNA-binding protein, IMP3 (also called IGF2BP3), which contains six RNA-binding domains (RBDs): four KH and two RRM domains. We establish an integrative systematic strategy, combining single-domain-resolved SELEX-seq, motif-spacing analyses, in vivo iCLIP, functional validation assays, and structural biology. This approach identifies the RNA-binding specificity and RNP topology of IMP3, involving all six RBDs and a cluster of up to five distinct and appropriately spaced CA-rich and GGC-core RNA elements, covering a >100 nucleotide-long target RNA region. Our generally applicable approach explains both specificity and flexibility of IMP3-RNA recognition, allows the prediction of IMP3 targets, and provides a paradigm for the function of multivalent interactions with multidomain RNA-binding proteins in gene regulation.enNamensnennung 4.0 Internationalddc:540