Cysteine proteases of Trypanosoma (Megatrypanum) theileri: cathepsin L-like gene sequences as targets for phylogenetic analysis, genotyping diagnosis.
Although Trypanosomatheileri and allied trypanosomes are the most widespread trypanosomes in bovids little is known about proteolytic enzymes in these species. We have characterized genes encoding for cathepsin L-like (CATL) cysteine proteases from isolates of cattle, water buffalo and deer that largely diverged from homologues of other trypanosome species. Analysis of 78 CATL catalytic domain sequences from 22 T. theileri trypanosomes disclosed 6 genotypes tightly clustered together into the T. theileri clade. The CATL genes in these trypanosomes are organized in tandem arrays of approximately 1.7kb located in 2 chromosomal bands of 600-720kb. A diagnostic PCR assay targeting CATL sequences detected T. theileri of all genotypes from cattle, buffaloes and cervids and also from tabanid vectors. Expression of T. theileri cysteine proteases was demonstrated by proteolytic activity in gelatin gels and hydrolysis of Z-Phe-Arg-AMC substrate. Results from this work agree with previous data using ribosomal and spliced leader genes demonstrating that CATL gene sequences are useful for diagnosis, population genotyping and evolutionary studies of T. theileri trypanosomes.
Rodrigues
Adriana C
AC
Departamento de Parasitologia, Universidade de São Paulo, São Paulo, SP, Brazil.
Garcia
Herakles A
HA
Ortiz
Paola A
PA
Cortez
Alane P
AP
Martinkovic
Franjo
F
Paiva
Fernando
F
Batista
Jael S
JS
Minervino
Antonio H
AH
Campaner
Marta
M
Pral
Elizabeth M
EM
Alfieri
Silvia C
SC
Teixeira
Marta M G
MM
D000595:Amino Acid Sequence; D000818:Animals; D002020:Buffaloes / Q000469:parasitology; D056668:Cathepsin L / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000469:parasitology; D057056:Cysteine Proteases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016054:DNA, Protozoan / Q000032:analysis / Q000235:genetics; D003670:Deer / Q000469:parasitology; D005838:Genotype; D058751:Livestock / Q000469:parasitology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D014345:Trypanosoma / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D014352:Trypanosomiasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*
D016054:DNA, Protozoan; D057056:Cysteine Proteases; D056668:Cathepsin L