Alternative splicing within and between alleles of the ATPase gene 1 locus of Trypanosoma brucei.

The P-type ATPase gene TBA1 of Trypanosoma brucei belongs to a polycistronic transcription unit. We analyzed the structure and expression of a 4-kb region located immediately downstream from TBA1. This region is unique and contains two large open reading frames transcribed into stable mRNAs. These putative genes, termed ADG1 and ADG2, can respectively encode a 24-kDa and a 81-kDa protein. The intergenic spacings between the polyadenylation sites and the next 3' splice acceptor sites are very short: 148 bp between TBA1 and ADG1, and 127 bp between ADG1 and ADG2. Transcripts from each of the two ADG1 alleles can be detected, indicating that both homologs are transcribed. These transcripts are differentially spliced due to a single base difference which destroys in one homolog the AG acceptor site present in the other. In the 'mutant' allele an alternative downstream splice acceptor site is used. Despite its sequence conservation in both alleles, this splice site is only used in the allele lacking the upstream AG acceptor site. The major population of ADG1 transcripts exhibit a long 5'-untranslated extension and no 3'-terminal tail, but a minor population shows a smaller 5'-untranslated region due alternative splicing closer to the initiation codon of the gene. The steady-state amounts of transcripts from individual genes in this region are differentially stage-regulated.