Solution Structure of the Second RRM Domain of RBM5 and Its Unusual Binding Characters for Different RNA Targets

The RNA binding motif protein 5 (RBMS), also known as LUCA15 or H37, containing two RNA recognition motifs, is a component of the spliceosome A complex. Previously, it has been reported that RBM5 bound to a U/C-rich sequence upstream of the In 100 element at intron 9 of caspase2 pre-mRNA that enhanced the formation of proapoptotic caspase-2L isoform. In the present study, we solved the solution structure of the RBM5 RRM2 core domain and characterized its unusual binding capability for different RNA sequences. We found that the RBMS RRM2 could preferentially bind to both CU rich and GA rich sequences with affinity in 10(-5) molar range. Further NMR experiments revealed that the dual RNA molecules could be accommodated on almost the same region of the protein's beta-sheet surface and that both the N- and C-terminal regions of the protein were involved in the recognition. Our studies provide evidence for the RBM5 sequence specific interaction with the cis-acting element in pre-mRNA regulating alternative splicing.