Synthesis and Spectroscopic Analysis of Branched RNA Fragments: Messenger RNA Splicing Intermediates

Article abstract of DOI:10.1021/jo00251a010

RNA splicing is now established as a major RNA processing reaction in eukaryotic cells.Splicing of messenger RNA precursors generates a lariat RNA structure containing a branched RNA core (Wallace J.C.; Edmons M.Proc.Natl.Acad.Sci.U.S.A. 1983, 80, 950-954) of the type A(2'p5'G)3'p5'X (A^G_X), where X is a pyrimidine residue.Understanding the mechanism which generates these molecules as well as the role they play in the splicing reaction are central issues in cell biology.Chemically synthesized branched RNA fragments of defined sequence and structure are likely to play a key role in understanding the full biological role of these molecules.Herein we describe the chemical synthesis of a series of trinucleotides (A^U_U, A^G_G, A^C_C, A^G_U, A^U_G, A^G_C, A^C_G, A^T_T, G^U_U, aA^U_U) and a tetranucleotide (UpA^U_U) with branched linkages.These molecules were fully characterized by UV, NMR (1H, 13C, 31P), HPLC analyses of enzymatic digests, and gel electrophoresis.