Indirect evidence suggests that the 5' end of U1 snRNA recognizes the 5' splice site in mRNA precursors by complementary base pairing. To test this hypothesis, we asked whether point mutations in the alternative 12S and 13S 5' splice sites of the adenovirus E1A gene can be suppressed by compensatory base changes in human U1 snRNA. When the mutant E1A and U1 genes are contransfected into HeLa cells, we observe efficient suppression of one mutation at position +5 in the 12S splice site, but exceedingly weak suppression of another mutation at position +3 in the 13S splice site. These and other results suggest that base pairing between U1 and the 5' splice site is necessary but not sufficient for the splicing of mRNA precursors.