The unusual 5' splicing border GC is used in myrosinase genes of the Brassicaceae

Abstract

Myrosinase (thioglucosidase glucohydrolase; EC 3.2.3.1) is a group of isoenzymes in the Brassicaceae, which hydrolyze glucosinolates. Genes encoding myrosinase contain 12 exons and 11 introns. Sequence comparison of two myrosinase genes from Arabidopsis thaliana, TGG1 and TGG2, with the corresponding cDNA from leaves, showed preferential use of a GC dinucleotide as the 5' splicing border in intron 1 instead of an adjacent GT dinucleotide four bp further 3'. This 5' GC splice site is conserved in all seven myrosinase genes characterized from three different species. Likewise, in the 3' region of intron 1 two AG dinucleotides are located seven bp apart. Only the most 5' of these dinucleotides was found to be used in splicing. Sequence analyses of TGG1 cDNA isolated from seeds, siliques and vegetative tissue using reverse transcription PCR showed that the splicing pattern of this intron is identical in these tissues for TGG1. The GT and the most 3' AG dinucleotides mentioned above have been assumed to be the intron borders of intron 1 in several myrosinase genes. The present investigation shows that this assumption is not correct.