Molecular characterization of a P-glycoprotein-related tcpgp2 gene in Trypanosoma cruzi.
We have cloned, sequenced and characterized a gene from Trypanosoma cruzi (Y strain), termed tcpgp2, which encodes a member of the ABC (ATP-binding cassette) superfamily of evolutionarily conserved transport proteins. The nucleotide sequence of the tcpgp2 gene was determined. It presents a 4602-bp open reading frame, coding for a 1534-amino acid protein, with a predicted molecular mass of 169,470 Da. The deduced amino acid sequence of tcpgp2 exhibited a remarkable homology with the P-glycoprotein-related genes of Leishmania tarentolae, the yeast cadmium factor (YCF1) and the human multidrug resistance-associated protein (MRP). Southern blot analysis using a specific probe indicated that the Tcpgp2 P-glycoprotein is encoded by a single copy gene which maps to a chromosome of about 900 kb. Northern blot analysis revealed that tcpgp2 gene is expressed as a polyadenylated transcript of approximately 5 kb in dividing amastigote and epimastigote forms; we did not detect the transcript in the non-dividing trypomastigote forms of the parasite. Gene transfection experiments in Leishmania tropica indicated that, under the conditions tested, tcpgp2 gene is not involved in drug resistance.
Dallagiovanna
B
B
Departamento de Bioquímica y Farmacología Molecular, Consejo Superiór de Investigaciones Científicas, Granada, Spain.
Gamarro
F
F
Castanys
S
S
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000737:chemistry*; D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D017125:Genes, Protozoan; D006801:Humans; D007891:Leishmania / Q000235:genetics; D007895:Leishmania tropica / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014162:Transfection; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000235:genetics*
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018528:ATP-Binding Cassette Transporters; D016054:DNA, Protozoan; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C102792:Tcpgp2 protein, Trypanosoma cruzi