Structural and functional analysis of an amplification containing a PGPA gene in a glucantime-resistant Leishmania (Viannia) guyanensis cell line.
Drug resistance is a complex phenomenon in Leishmania and commonly involves gene amplification. Active efflux and metal sequestration through a P-glycoprotein have been pointed to as the major mechanisms used by drug-resistant Leishmania. A gene amplification from a glucantime-resistant Leishmania (Viannia) guyanensis cell line was characterised in an attempt to understand the mechanism of metal resistance in this pathogenic species. We show that the amplification is present as an extrachromosomal amplicon of 30 kb and contains a PGPA gene ( LgPGPA), which is overexpressed in the resistant line as shown by Northern and Western blot analyses. In addition, we gathered evidence from transfection experiments for the role of the LgPGPA gene in oxyanion resistance in L. (V.) guyanensis. Our work indicates that, in this pathogenic New World Leishmania species, amplification of the PGPA gene is the major determinant in oxyanion resistance.
Anacleto
Charles
C
Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP 486, CEP 31270-901, Belo Horizonte, MG, Brazil.
Abdo
Maria C B
MC
Ferreira
Adlane V B
AV
Murta
Silvane M F
SM
Romanha
Alvaro J
AJ
Fernandes
Ana Paula
AP
Moreira
Elizabeth S A
ES
D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D004351:Drug Resistance; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D005784:Gene Amplification; D005786:Gene Expression Regulation; D017125:Genes, Protozoan; D018334:Leishmania guyanensis / Q000187:drug effects / Q000235:genetics*; D008536:Meglumine / Q000494:pharmacology*; D000077485:Meglumine Antimoniate; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009942:Organometallic Compounds / Q000494:pharmacology*; D015800:Protozoan Proteins; D015183:Restriction Mapping; D016415:Sequence Alignment; D014162:Transfection
D018528:ATP-Binding Cassette Transporters; D008562:Membrane Glycoproteins; D009942:Organometallic Compounds; C106891:PgpA protein, Leishmania; D015800:Protozoan Proteins; D008536:Meglumine; D000077485:Meglumine Antimoniate