The AG Dinucleotide Terminating Introns Is Important but Not Always Required for Pre-mRNA Splicing in the Maize Endosperm1

Abstract

Abstract Previous RNA analysis of lesions within the 15 intron-containingSh2(shrunken2) gene of maize (Zea mays) revealed that the majority of these mutants affect RNA splicing. Here we decipher further two of these mutants, sh2-i(shrunken2intermediate phenotype) andsh2-7460. Each harbors a G-to-A transition in the terminal nucleotide of an intron, hence destroying the invariant AG found at the terminus of virtually all nuclear introns. Consequences of the mutations, however, differ dramatically. In sh2-i the mutant site is recognized as an authentic splice site in approximately 10% of the primary transcripts processed in the maize endosperm. The other transcripts exhibited exon skipping and lacked exon 3. A G-to-A transition in the terminus of an intron was also found in the mutantsh2-7460, in this case intron 12. The lesion activates a cryptic acceptor site downstream 22 bp within exon 13. In addition, approximately 50% ofsh2-7460 transcripts contain intron 2 and 3 sequences.