Journal article

A systems view of spliceosomal assembly and branchpoints with iCLIP

Michael Briese, Nejc Haberman, Christopher Sibley, Anob Chakrabarti, Zhen Wang, Julian König, David Perera, Vihandha Wickramasinghe, Ashok Venkitaraman, Nicholas Luscombe, Christopher Smith, Tomaž Curk, Jernej Ule

Published : 2018


Studies of spliceosomal interactions are challenging due to their dynamic nature. Here we employed spliceosome iCLIP, which immunoprecipitates SmB along with snRNPs and auxiliary RNA binding proteins (RBPs), to map human spliceosome engagement with snRNAs and pre-mRNAs. This identified over 50,000 branchpoints (BPs) that have canonical sequence and structural features. Moreover, it revealed 7 binding peaks around BPs and splice sites, each precisely overlapping with binding profiles of specific splicing factors. We show how the binding patterns of these RBPs are affected by the position and strength of BPs. For example, strong or proximally located BPs preferentially bind SF3 rather than U2A..

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