Reversibly constraining spliceosome-substrate complexes by engineering disulfide crosslinks

Engineering Crystal Packing in RNA-Protein Complexes II: A Historical Perspective from the Structural Studies of the Spliceosome

“…The structure also explained how U1C stabilizes the interaction between a 5′ss and U1 snRNA and how U1-70K facilitates this interaction via its long unstructured N-terminus. The structure was of such high quality that it was possible to use it to guide the engineering of a disulfide cross-link between a 5′ss nucleoside and a proximal cysteine in U1-C [ 32 ].…”

Structural and functional analyses of the spliceosome requires a multi-disciplinary approach

“…MacMillan and colleagues describe the methods used to synthesize a directed hydroxyl radical probe tethered to an mRNA substrate that can be used to probe the structures sampled during the spliceosome assembly process [41]. Dr. Pomeraz Krummel and colleagues present a research article on methods that can be used to covalently and reversibly capture the spliceosome at specific stages of spliceosome function [42]. Fairbrother and colleagues discuss the importance of pre-mRNA structure in the splicing reaction and the challenges associated with predicting RNA structure using computational approaches [43].…”