A U1 snRNA:pre‐mRNA base pairing interaction is required early in yeast spliceosome assembly but does not uniquely define the 5′ cleavage site.

We analyzed the effects of suppressor mutations in the U1 snRNA (SNR19) gene from Saccharomyces cerevisiae on the splicing of mutant pre‐mRNA substrates. The results indicate that pairing between U1 snRNA and the highly conserved position 5 (GTATGT) of the intron occurs early in spliceosome assembly in vitro. This pairing is important for efficient splicing both in vitro and in vivo. However, pairing at position 5 does not appear to influence 5′ splice site selection in vivo, indicating that the previously described U1 snRNA:5′ splice junction base pairing interaction is not sufficient to define the 5′ cleavage site.