Trypanosoma cruzi: allelic comparisons of the actin genes and analysis of their transcripts

Abstract

Two allelic genomic fragments containing actin genes from Trypanosoma cruzi were isolated and characterized. One allele comprises two complete tandem repeats of a sequence encoding an actin gene. In the other, only one actin gene is found. Each one of these three gene copies encode for a complete and identical potential protein of 376 amino acids which is 93% similar with its homolog from Trypanosoma brucei. Northern hybridizations of both total and polysomal RNA from epimastigotes demonstrated the presence of an actin polyadenylated mRNA of about 1.6 kb. Actin transcripts processing sites were determined by 5 ′- and 3 ′-RACE. The obtained sequence data demonstrates that actin genes from both alleles are expressed. The stability of actin mRNA was found to be similar to the one exhibited by the ribosomal protein S4 mRNA as an internal reference. A time course analysis of cultured epimastigotes showed a novel behaviour in which actin mRNA steady state concentration peaks during the transition from the logarithmic to the stationary phase of growth. Index Descriptors and Abbreviations: cDNA, complementary DNA; EDTA, ethylenediaminetetraacetic acid; mRNA, messenger RNA; ORF, open reading frame; PCR, polymerase chain reaction; RACE, rapid amplification of cDNA ends; RNase, ribonuclease; SIRE, short interspersed element; ssrRNA, small subunit ribosomal RNA; UV, ultraviolet; UTR, untranslated region