Overexpression of Ubiquitin and Amino Acid Permease Genes in Association with Antimony Resistance in Leishmania tropica Field Isolates

• Published in: Korean journal of parasitology 2013-08, Vol.51 (4), p.413-419
• Main Authors: Kazemi-Rad, E., Tehran University of Medical Sciences, Tehran, Iran, Mohebali, M., Tehran University of Medical Sciences, Tehran, Iran, Khadem-Erfan, M.B., Tehran University of Medical Sciences, Tehran, Iran, Hajjaran, H., Tehran University of Medical Sciences, Tehran, Iran, Hadighi, R., Iran University of Medical Sciences, Tehran, Iran, Khamesipour, A., Tehran University of Medical Sciences, Tehran, Iran, Rezaie, S., Tehran University of Medical Sciences, Tehran, Iran, Saffari, M., Tehran University of Medical Sciences, Tehran, Iran, Raoofian, R., Tehran University of Medical Sciences, Tehran, Iran, Heidari, M., Tehran University of Medical Sciences, Tehran, Iran
• Format: Article
• Language: English
• Subjects: Amino Acid Transport Systems - genetics; Amino Acid Transport Systems - metabolism; Antimony - pharmacology; antimony resistance, cDNA-AFLP, real-time RT-PCR, ubiquitin, amino acid permease; Antipruritics - pharmacology; Drug Resistance; Humans; LEISHMANIA TROPICA; Leishmania tropica - drug effects; Leishmania tropica - enzymology; Leishmania tropica - genetics; Leishmania tropica - isolation & purification; Leishmaniasis, Cutaneous - parasitology; Original; Protozoan Proteins - genetics; Protozoan Proteins - metabolism; Ubiquitin - genetics; Ubiquitin - metabolism
• ISSN: 0023-4001; 1738-0006; 1738-0006
• DOI: 10.3347/kjp.2013.51.4.413

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) (P0.05) 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.