Browse > Article
http://dx.doi.org/10.3347/kjp.2013.51.4.413

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

Kazemi-Rad, Elham (Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences)
Mohebali, Mehdi (Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences)
Erfan, Mohammad Bagher Khadem (Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences)
Hajjaran, Homa (Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences)
Hadighi, Ramtin (Department of Medical Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences)
Khamesipour, Ali (Center for Research & Training in Skin Diseases & Leprosy, Tehran University of Medical Sciences)
Rezaie, Sassan (Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences)
Saffari, Mojtaba (Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences)
Raoofian, Reza (Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences)
Heidari, Mansour (Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences)
Publication Information
Parasites, Hosts and Diseases / v.51, no.4, 2013 , pp. 413-419 More about this Journal
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$^{(R)}$) 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 resistant isolates compared to sensitive ones. Our results suggest that overexpression of ubiquitin and AAP3 could potentially implicated in natural antimony resistance.
Keywords
Leishmania tropica; antimony resistance; cDNA-AFLP; real-time RT-PCR; ubiquitin; amino acid permease;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Leonard SS, Harris GK, Shi X. Metal-induced oxidative stress and signal transduction. Free Radic Biol Med 2004; 37: 1921-1942.   DOI   ScienceOn
2 Akerman M, Shaked-Mishan P, Mazareb S, Volpin H, Zilberstein D. Novel motifs in amino acid permease genes from Leishmania. Biochem Biophys Res Commun 2004; 325: 353-366.   DOI   ScienceOn
3 Klemperer N, Pickart C. Arsenite inhibits two steps in the ubiquitin- dependent proteolytic pathway. J Biol Chem 1989; 264: 19245-19252.
4 Cheng L, Watt R, Piper P. Polyubiquitin gene expression contributes to oxidative stress resistance in respiratory yeast (Saccharomyces cerevisiae). Mol Gen Genet 1994; 243: 358-362.   DOI
5 Friant S, Meier KD, Riezman H. Increased ubiquitin-dependent degradation can replace the essential requirement for heat shock protein induction. EMBO J 2003; 22: 3783-3791.   DOI   ScienceOn
6 Shaked-Mishan P, Suter-Grotemeyer M, Yoel-Almagor T, Holland N, Zilberstein D, Rentsch D. A novel high-affinity arginine transporter from the human parasitic protozoan Leishmania donovani. Mole Microbiol 2006; 60: 30-38.   DOI   ScienceOn
7 Colotti G, Ilari A. Polyamine metabolism in Leishmania: from arginine to trypanothione. Amino Acids 2011; 40: 269-285.   DOI
8 Fairlamb AH, Cerami A. Metabolism and functions of trypanothione in the kinetoplastida. Annu Rev Microbiol 1992; 46: 695- 729.   DOI   ScienceOn
9 Mukhopadhyay R, Dey S, Xu N, Gage D, Lightbody J, Ouellette M, Rosen BP. Trypanothione overproduction and resistance to antimonials and arsenicals in Leishmania. Proc Natl Acad Sci USA 1996; 93: 10383-10387.   DOI   ScienceOn
10 Sundar S. Drug resistance in Indian visceral leishmaniasis. Trop Med Int Health 2002; 6: 849-854.
11 Murray HW, Berman JD, Davies CR, Saravia NG. Advances in leishmaniasis. Lancet 2005; 366: 1561-1577.   DOI   ScienceOn
12 Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, Cano J, Jannin J, Boer Md, the WHOLCT. Leishmaniasis worldwide and global estimates of its incidence. PLoS One 2012; 7: e35671.   DOI
13 Statistics of Cutaneous Leishmanaisis in Iran: Hearing before National Leishmaniasis Committee, Office of Zoonoses, Diseases Management Center, Ministry of Health and Medical Education (2004).
14 Talari SA, Talaei R, Shajari G, Vakili Z, Taghaviardakani A. Childhood cutaneous leishmaniasis: report of 117 cases from Iran. Korean J Parasitol 2006; 44: 355-360.   DOI   ScienceOn
15 Kedzierski L, Sakthianandeswaren A, Curtis JM, Andrews PC, Junk PC, Kedzierska K. Leishmaniasis: current treatment and prospects for new drugs and vaccines. Curr Med Chem 2009; 16: 599-614.   DOI   ScienceOn
16 Marquis N, Gourbal B, Rosen BP, Mukhopadhyay R, Ouellette M. Modulation in aquaglyceroporin AQP1 gene transcript levels in drug-resistant Leishmania. Mol Microbiol 2005; 57: 1690-1699.   DOI   ScienceOn
17 Hadighi R, Mohebali M, Boucher P, Hajjaran H, Khamesipour A, Ouellette M. Unresponsiveness to glucantime treatment in Iranian cutaneous leishmaniasis due to drug-resistant Leishmania tropica parasites. PLoS Med 2006; 3: e162.   DOI
18 Ashutosh, Sundar S, Goyal N. Molecular mechanisms of antimony resistance in Leishmania. J Med Microbiol 2007; 56: 143- 153.   DOI   ScienceOn
19 Haimeur A, Guimond C, Pilote S, Mukhopadhyay R, Rosen BP, Poulin R, Ouellette M. Elevated levels of polyamines and trypanothione resulting from overexpression of the ornithine decarboxylase gene in arsenite-resistant Leishmania. Mol Microbiol 2002; 34: 726-735.
20 Walker J, Gongora R, Vasquez JJ, Drummelsmith J, Burchmore R, Roy G, Ouellette M, Gomez MA, Saravia NG. Discovery of factors linked to antimony resistance in Leishmania panamensis through differential proteome analysis. Mol Biochem Parasitol 2012; 183: 166-176.   DOI   ScienceOn
21 El Fadili K, Messier N, Leprohon P, Roy G, Guimond C, Trudel N, Saravia NG, Papadopoulou B, Legare D, Ouellette M. Role of the ABC transporter MRPA (PGPA) in antimony resistance in Leishmania infantum axenic and intracellular amastigotes. Antimicrob Agents Chemother 2005; 49: 1988-1993.   DOI   ScienceOn
22 Vergnes B, Gourbal B, Girard I, Sundar S, Drummelsmith J, Ouellette M. A proteomics screen implicates HSP83 and a small kinetoplastid calpain-related protein in drug resistance in Leishmania donovani clinical field isolates by modulating drug-induced programmed cell death. Mol Cell Proteomics 2007; 6: 88-101.
23 Do Monte-Neto RL, Coelho AC, Raymond F, Legare D, Corbeil J, Melo MN, Frezard F, Ouellette M. Gene expression profiling and molecular characterization of antimony resistance in Leishmania amazonensis. PLoS Negl Trop Dis 2011; 5: e1167.   DOI
24 Hajjaran H, Azarian B, Mohebali M, Hadighi R, Assareh A, Vaziri B. Comparative proteomics study on meglumine antimoniate sensitive and resistant Leishmania tropica isolated from Iranian anthroponotic cutaneous leishmaniasis patients. East Mediterr Health J 2012; 18: 165-171.
25 Saffari M, Dinehkabodi OS, Ghaffari SH, Modarressi MH, Mansouri F, Heidari M. Identification of novel p53 target genes by cDNA AFLP in glioblastoma cells. Cancer Lett 2009; 273: 316- 322.   DOI   ScienceOn
26 Vuylsteke M, Peleman JD, van Eijk MJ. AFLP-based transcript profiling (cDNA-AFLP) for genome-wide expression analysis. Nat Protoc 2007; 2: 1399-1413.   DOI   ScienceOn
27 Farahyar S, Zaini F, Kordbacheh P, Rezaie S, Safara M, Raoofian R, Heidari M. Overexpression of aldo-keto-reductase in azole-resistant clinical isolates of Candida glabrata determined by cDNAAFLP. DARU 2013; 21: 1-7.   DOI   ScienceOn
28 Sherman MY, Goldberg AL. Cellular defenses against unfolded proteins: A cell biologist thinks about neurodegenerative diseases. Neuron 2001; 29: 15-32.   DOI   ScienceOn
29 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-deltadeltact method. Methods 2001; 25: 402-408.   DOI   ScienceOn
30 Welchman RL, Gordon C, Mayer RJ. Ubiquitin and ubiquitinlike proteins as multifunctional signals. Nat Rev Mol Cell Biol 2005; 6: 599-609.   DOI   ScienceOn
31 Ciechanover A. Intracellular protein degradation: from a vague idea, through the lysosome and the ubiquitin-proteasome system, and onto human diseases and drug targeting (nobel lecture). Angew Chem Int Ed Engl 2005; 44: 5944-5967.   DOI   ScienceOn
32 Tsirigotis M, Zhang M, Chiu RK, Wouters BG, Gray DA. Sensitivity of mammalian cells expressing mutant ubiquitin to proteindamaging agents. J Biol Chem 2001; 276: 46073-46078.   DOI   ScienceOn
33 Fujimuro M, Nishiya T, Nomura Y, Yokosawa H. Involvement of polyubiquitin chains via specific chain linkages in stress response in mammalian cells. Biol Pharm Bull 2005; 28: 2315-2318.   DOI   ScienceOn
34 Del Razo LM, Quintanilla-Vega B, Brambila-Colombres E, Calderón- Aranda ES, Manno M, Albores A. Stress proteins induced by arsenic. Toxicol Appl Pharmacol 2001; 177: 132-148.   DOI   ScienceOn