title: Structural and functional analysis of an amplification containing a PGPA gene in a glucantime-resistant Leishmania (Viannia) guyanensis cell line.
abstract: 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.
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP 486, CEP 31270-901, Belo Horizonte, MG, Brazil.
authors.forename: Charles
authors.initials: C
authors.lastname: Anacleto
authors.forename: Maria C B
authors.initials: MC
authors.lastname: Abdo
authors.forename: Adlane V B
authors.initials: AV
authors.lastname: Ferreira
authors.forename: Silvane M F
authors.initials: SM
authors.lastname: Murta
authors.forename: Alvaro J
authors.initials: AJ
authors.lastname: Romanha
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Fernandes
authors.forename: Elizabeth S A
authors.initials: ES
authors.lastname: Moreira
mesh_terms: 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
chemical_list: D018528:ATP-Binding Cassette Transporters; D008562:Membrane Glycoproteins; D009942:Organometallic Compounds; C106891:PgpA protein, Leishmania; D015800:Protozoan Proteins; D008536:Meglumine; D000077485:Meglumine Antimoniate