Microbial iCLIP2: enhanced mapping of RNA–protein interaction by promoting protein and RNA stability

Adaption and improvement of iCLIP2 procedure. (A) Diagrammatic overview of iCLIP2 procedure, with a specific emphasis on cell lysis (marked). (B) Endosomal mRNA transport model in U. maydis. The RBP Rrm4 (RNA recognition motif protein 4, petrol) binds directly to cargo mRNAs (black) and cotransports the associated RBPs like Grp1 (glycine-rich protein 1, mint) or the poly(A)-binding protein (Pab1, blue) in an RNA-dependent manner. (C) Schematic overview of the size (in AA) and domain architecture ([RRM] RNA recognition motif; [MLLE] MademoiseLLE domain) of Rrm4, the amino acid substitutions mutant Rrm4^mR1 and the RBP Grp1. (D) Western blot analysis of immunoprecipitated Rrm4-G (Rrm4-G; 112 kDa).