title: Overexpression of ubiquitin and amino acid permease genes in association with antimony resistance in Leishmania tropica field isolates.
abstract: The mainstay therapy against leishmaniasis is still pentavalent antimonial drugs; however, the rate of antimony resistance is increasing in endemic regions such as Iran. Understanding the molecular basis of resistance to antimonials could be helpful to improve treatment strategies. This study aimed to recognize genes involved in antimony resistance of Leishmania tropica field isolates. Sensitive and resistant L. tropica parasites were isolated from anthroponotic cutaneous leishmaniasis patients and drug susceptibility of parasites to meglumine antimoniate (Glucantime®) was confirmed using in vitro assay. Then, complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) and real-time reverse transcriptase-PCR (RT-PCR) approaches were utilized on mRNAs from resistant and sensitive L. tropica isolates. We identified 2 known genes, ubiquitin implicated in protein degradation and amino acid permease (AAP3) involved in arginine uptake. Also, we identified 1 gene encoding hypothetical protein. Real-time RT-PCR revealed a significant upregulation of ubiquitin (2.54-fold), and AAP3 (2.86-fold) (P<0.05) in a resistant isolate compared to a sensitive one. Our results suggest that overexpression of ubiquitin and AAP3 could potentially implicated in natural antimony resistance.
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Elham
authors.initials: E
authors.lastname: Kazemi-Rad
authors.forename: Mehdi
authors.initials: M
authors.lastname: Mohebali
authors.forename: Mohammad Bagher
authors.initials: MB
authors.lastname: Khadem-Erfan
authors.forename: Homa
authors.initials: H
authors.lastname: Hajjaran
authors.forename: Ramtin
authors.initials: R
authors.lastname: Hadighi
authors.forename: Ali
authors.initials: A
authors.lastname: Khamesipour
authors.forename: Sassan
authors.initials: S
authors.lastname: Rezaie
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Saffari
authors.forename: Reza
authors.initials: R
authors.lastname: Raoofian
authors.forename: Mansour
authors.initials: M
authors.lastname: Heidari
mesh_terms: D026905:Amino Acid Transport Systems / Q000235:genetics* / Q000378:metabolism; D000965:Antimony / Q000494:pharmacology*; D000982:Antipruritics / Q000494:pharmacology*; D004351:Drug Resistance; D006801:Humans; D007895:Leishmania tropica / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology*; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D025801:Ubiquitin / Q000235:genetics* / Q000378:metabolism
chemical_list: D026905:Amino Acid Transport Systems; D000982:Antipruritics; D015800:Protozoan Proteins; D025801:Ubiquitin; D000965:Antimony
keywords: Leishmania tropica; amino acid permease; antimony resistance; cDNA-AFLP; real-time RT-PCR; ubiquitin
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