Abstract
Four Trypanosoma cruzi genomic DNA fragments carrying different TcP2β genes have been isolated and sequenced. Three of them had a single TcP2β gene, while the 3.8-kb-long DNA segment encoding the TcP2β-H1.8 locus showed two TcP2β genes arranged in tandem. These genes were physically connected by a 428-bp-long DNA sequence that was also located immediately 5′ to the first gene and immediately 3′ to the second. Comparison of the 4 TcP2β gene loci, suggested that the insertion of this repeated element originated the duplication of its target sequence, a poly(dT) stretch. Approximately 1200 copies of this short sequence, named short interspersed repetitive element (SIRE), were found scattered in the genome. Analysis of the 5′ non-coding regions of different TcP2β mRNAs, and RNA-PCR experiments suggested that the insertion of a SIRE upstream of a TcP2β-H 1.8 gene introduced a new 3′ spliced leader (SL) acceptor site in the TcP2β-H1.8 pre-mRNAs, encoded within the SIRE. Consequently, in the mature H1.8 mRNA the SL sequence is followed by 38 bases directly transcribed from the SIRE. Structural and functional features of this repeated element reveal similarity to the short interspersed repetitive DNA sequences detected in the genomes of several microorganisms.
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