Bimolecular Exon Ligation by the Human Spliceosome

Abstract

Intron excision is an essential step in eukaryotic gene expression, but the molecular mechanisms by which the spliceosome accurately identifies splice sites in nuclear precursors to messenger RNAs (pre-mRNAs) are not well understood. A bimolecular assay for the second step of splicing has now revealed that exon ligation by the human spliceosome does not require covalent attachment of a 3′ splice site to the branch site. Furthermore, accurate definition of the 3′ splice site in this system is independent of either a covalently attached polypyrimidine tract or specific 3′ exon sequences. Rather, in this system 3′ splice site selection apparently occurs with a 5′ → 3′ directionality.