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KMID : 0425120130510040413
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Parasites, Hosts and Diseases
2013 Volume.51 No. 4 p.413 ~ p.419
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Overexpression of Ubiquitin and Amino Acid Permease Genes in Association with Antimony Resistance in Leishmania tropica Field Isolates
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Kazemi-Rad Elham
Mohebali Mohebali
Khadem-Erfan Mohammad Bagher
Hajjaran Homa
Hadighi Ramtin
Khamesipour Ali
Rezaie Sassan
Saffari Mojtaba
Raoofian Reza
Heidari Mansour
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Abstract
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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.
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KEYWORD
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Leishmania tropica, antimony resistance, cDNA-AFLP, real-time RT-PCR, ubiquitin, amino acid permease
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