Trypanosoma rangeli and Trypanosoma cruzi: molecular characterization of genes encoding putative calcium-binding proteins, highly conserved in trypanosomatids.
Genes encoding a 29-kDa flagellar calcium-binding protein (F29) in Trypanosoma cruzi, strongly homologous to EF-hand calcium-binding protein-encoding genes previously reported in this parasite, were isolated by immunoscreening. F29 is encoded by a number of very similar genes, highly conserved among different T. cruzi isolates. The genes are located on a pair of homologous chromosomes, arranged in one or two clusters of tandem repeats. PCR amplification of Trypanosoma rangeli genomic DNA, using primers derived from the T. cruzi F29 sequence made it possible to isolate the homologous gene in T. rangeli, encoding a 23-kDa protein called TrCaBP. Gene sequence comparisons showed homology to EF-hand calcium-binding proteins from T. cruzi (82.8%), Trypanosoma brucei brucei (60.2%), and Entamoeba histolytica (28.4%). Northern blot analysis revealed that the TrCaBP gene is expressed in T. rangeli as a polyadenylated transcript. The TrCaBP-encoding genes are present in at least 20 copies per cell, organized in tandem arrays, on large T. rangeli chromosomes in some isolates and on two smaller ones in others. This gene, however, seems to be absent from Leishmania.
Porcel
B M
BM
Instituto Nacional de Chagas, Ministerio de Salud y Acción Social, Buenos Aires, Argentina.
Bontempi
E J
EJ
Henriksson
J
J
Rydåker
M
M
Aslund
L
L
Segura
E L
EL
Pettersson
U
U
Ruiz
A M
AM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002135:Calcium-Binding Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D017124:Conserved Sequence; D016054:DNA, Protozoan / Q000737:chemistry; D005407:Flagella / Q000737:chemistry; D008969:Molecular Sequence Data; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D016053:RNA, Protozoan / Q000032:analysis; D012091:Repetitive Sequences, Nucleic Acid; D016415:Sequence Alignment; D014345:Trypanosoma / Q000235:genetics* / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000235:genetics* / Q000378:metabolism
D002135:Calcium-Binding Proteins; D016054:DNA, Protozoan; D015800:Protozoan Proteins; D016053:RNA, Protozoan; C061478:flagellar calcium-binding protein, Trypanosoma cruzi