Comparison and Evolutionary Analysis of the Glycosomal Glyceraldehyde-3-Phosphate Dehydrogenase from Different Kinetoplastida

Abstract

In this work, we present the sequences and a comparison of the glycosomal GAPDHs from a number of Kinetoplastida. The complete gene sequences have been determined for some species (Crithidia fasciculata, Herpetomonas samuelpessoai, Leptomonas seymouri, and Phytomonas sp), whereas for other species (Trypanosoma brucei gambiense, Trypanosoma congolense, Trypanosoma vivax, and Leishmania major), only partial sequences have been obtained by PCR amplification. The structure of all available glycosomal GAPDH genes was analyzed in detail. Considerable variations were observed in both their nucleotide composition and their codon usage. The GC content varies between 64.4% in L. seymouri and 49.5% in the previously sequenced GAPDH gene from Trypanoplasma borreli. A highly biased codon usage was found in C. fasciculata, with only 34 triplets used, whereas in T. borreli 57 codons were employed. No obvious correlation could be observed between the codon usage and either the nucleotide composition or the level of gene expression. The glycosomal GAPDH is a very well-conserved enzyme. The maximal overall difference observed in the amino acid sequences is only 25%. Specific insertions and extensions are retained in all sequences. The residues involved in catalysis, substrate, and inorganic phosphate binding are fully conserved, whereas some variability is observed in the cofactor-binding pocket. The implications of these data for the design of new trypanocidal drugs targeted against GAPDH are discussed. All available gene and amino acid sequences of glycosomal GAPDHs were used for a phylogenetic analysis. The division of the Kinetoplastida into two suborders, Bodonina and Trypanosomatina, was well supported. Within the letter group, the Trypanosoma species appeared to be monophyletic, whereas the other trypanosomatids form a second clade.

Phylogeny, taxonomy and grouping of Trypanosoma rangeli isolates from man, triatomines and sylvatic mammals from widespread geographical origin based on SSU and ITS ribosomal sequences

“…The genus Trypanosoma seems to be among the most diverse lineage within the Trypanosomatidae family and its monophyly has been supported by several studies (Haag, O'h Uigin & Overath, 1998 ;Hannaert, Opperdoes & Michels, 1998 ;Stevens et al b, 2001. Molecular phylogenetic studies supported the division of mammalian trypanosomes into 2 major groups, which are represented by stercorarian or salivarian trypanosomes, separated by a very large genetic distance and constituted by highly divergent subgenera.…”

Kinetoplastid phylogenetics, with special reference to the evolution of parasitic trypanosomes

“…E-mail: j.r.stevens@ex.ac.uk 2000) and even in those parasites of medical importance, it is little more than a decade since the widespread introduction of DNA sequencing technologies has allowed the evolution of parasitic trypanosomatids (and kinetoplastids in general) to be evaluated by formal phylogenetic analysis (Fernandes et al, 1993;Alvarez et al, 1996;Maslov et al, 1996;Lukes et al, 1997;Haag et al, 1998;Hannaert et al, 1998;Stevens et al, 1999Stevens et al, , 2001Wright et al, 1999;Hamilton et al, 2004;Moreira et al, 2004). To fully understand the evolutionary history of parasitic kinetoplastids and to understand the context within which the evolution of each parasite group has unfolded, an understanding not just of the parasites, but also of broader kinetoplastid phylogenetics is required.…”

“…Confidence in 18S rDNA sequence data to infer phylogenetic relationships has also been challenged by the observation of two distinct copies of these genes in T. cruzi and other trypanosomatids (Stothard et al, 1998). In contrast, phylogenetic studies based on proteincoding genes (Hannaert et al, 1992(Hannaert et al, , 1998Hashimoto et al, 1995;Adjé et al, 1998;Simpson et al, 2002) have consistently and unequivocally supported monophyly, a conclusion strongly supported by Hamilton et al (2004) in their recent analyses of both glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) DNA and protein trees from a broad range of trypanosomatid taxa, rooted using Euglena gracilis, an unequivocal outgroup taxon. Trypanosome monophyly is further supported by the phylogenies of Simpson et al (2002) based on heat shock protein 90 amino acid sequences that include a different range of kinetoplastid taxa to the gGAPDH gene trees.…”

Genes de cisteíno proteases (Catepsina L-like) de Trypanosoma rangeli: polimorfismo, relações filogenéticas e alvos para diagnóstico e genotipagem.

“…Estes estudos incluíram análises dos genes gGAPDH, Fator de elongação 1α, tripanotiona redutase, β tubulina, HSP90 e HSP70, que geraram filogenias congruentes em apresentar aos tripanossomas como um grupo monofilético (HASHIMOTO et al, 1995;ALVAREZ et al, 1996;HANNAERT et al, 1998;LUKES et al, 2002;HAMILTON et al, 2007).…”