- INTRODUCTION
- MATERIALS AND METHODS
- Parasite and culture condition.
- Chemosensitivity profiles of SAG-S and SAG-R promasti-gotes to potassium antimony tartrate (SbIII).
- Uptake assay.
- Cloning of full-length aquaglyceroporin (AQP1) gene from antimony-sensitive and antimony-resistant L. donovani strains.
- Nucleic acid isolation and hybridization analysis.
- RESULTS
- Susceptibility of Leishmania isolates to potassium antimony tartrate (SbIII).
- Comparison of transport of potassium antimony tartrate (SbIII) in SAG-R versus SAG-S field isolates.
- Southern blot analysis of the aquaglyceroporin (AQP1) gene in SAG-S and SAG-R field isolates.
- Comparison of aquaglyceroporin gene expression in SAG-S versus SAG-R field isolates.
- DISCUSSION
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Reference Manager
BibTeX
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-
1
Guerin PJ, Olliaro P, Sundar S, Boelaert M, Croft SL, Desjeux P, Wasunna MK, Bryceson ADM, 2002. Visceral leishmaniasis: current status of control, diagnosis, and treatment, and a proposed research and development agenda. Lancet Infect Dis 2 :494–501.
-
2
Roberts WL, Berman JD, Rainey PM, 1995. In vitro antileishmanial properties of tri- and pentavalent antimonial preparations. Antimicrob Agents Chemother 39 :1234–1239.
-
3
Shaked-Mishan P, Ulrich N, Ephros M, Zilberstein D, 2001. Novel intracellular SbV reducing activity correlates with antimony susceptibility in Leishmania donovani. J Biol Chem 276 :3971–3976.
-
4
Singh AK, Liu HY, Lee ST, 1994. Atomic absorption spectro-photometric measurement of intracellular arsenite in arsenite-resistant Leishmania. Mol Biochem Parasitol 66 :161–164.
-
5
Denton H, McGregor JC, Coombs GH, 2004. Reduction of anti-leishmanial pentavalent antimonial drugs by a parasite-specific thiol-dependent reductase, TDR1. Biochem J 381 :405–412.
-
6
Zhou Y, Messier N, Ouellette M, Rosen BP, Mukhopadhyay R, 2004. Leishmania major LmACR2 is a pentavalent antimony reductase that confers sensitivity to the drug pentostam. J Biol Chem 279 :37445–37451.
-
7
Sereno D, Cavaleyra M, Zemzoumi K, Maquaire S, Ouaissi A, Lemesre JL, 1998. Axenically grown amastigotes of Leishmania infantum used as an in vitro model to investigate the pentavalent antimony mode of action. Antimicrob Agents Chemother 42 :3097–3102.
-
8
Sundar S, More DK, Singh MK, Singh VP, Sharma S, Makharia A, Kumar PCK, Murray W, 2000. Failure of pentavalent antimony in visceral leishmaniasis in India: report from the centre of the Indian epidemic. Clin Infect Dis 31 :1104–1107.
-
9
Lira R, Sundar S, Makharia A, Kenney R, Gam A, Saraiva E, Sacks D, 1999. Evidence that the high incidence of treatment failures in Indian kala-azar is due to the emergence of antimony-resistant strains of Leishmania donovani. J Infect Dis 180 :564–567.
-
10
Sundar S, 2001. Drug resistant in Indian Visceral leishmaniasis. Trop Med Int Health 6 :849–854.
-
11
Mukhopadhyay R, Dey S, Xu N, Gage D, Lightbody J, Ouellette M, Rosen BP, 1996. Trypanothione overproduction and resistance to antimonials and arsenicals in Leishmania. Proc Natl Acad Sci USA 93 :10383–10387.
-
12
Haimeur A, Guimond C, Pilote S, Mukhopadhyay R, Rosen BP, Poulin R, Ouellette M, 1999. Elevated levels of polyamines and trypanothione resulting from overexpression of the ornithine decarboxylase gene in arsenite-resistant Leishmania. Mol Microbiol 34 :726–735.
-
13
Haimeur A, Brochu C, Genest PA, Papadopoulou B, Ouellette M, 2000. Amplification of the ABC transporter gene PGPA and increased trypanothione levels in potassium antimonyl tartrate (SbIII) resistant Leishmania tarentolae. Mol Biochem Parasitol 108 :131–135.
-
14
Legare D, Papadopoulou B, Roy G, Mukhopadhyay R, Haimeur A, Dey S, Grondin K, Brochu C, Rosen BP, Ouellette M, 1997. Efflux systems and increased trypanothione levels in arsenite-resistant Leishmania. Exp Parasitol 87 :275–282.
-
15
Legare D, Richard D, Mukhopadhyay R, Stierhof YD, Rosen BP, Haimeur A, Papadopoulou B, Ouellette M, 2001. The Leishmania ABC protein PGPA is an intracellular metal-thiol transporter ATPase. J Biol Chem 276 :26301–26307.
-
16
Mukherjee A, Padmanabhan PK, Singh S, Roy G, Girard I, Chatterjee M, Ouellette M, Madhubala R, 2007. Role of ABC transporter MRPA, γ-glutamylcysteine synthetase and ornithine decarboxylase in natural antimony-resistant isolates of Leishmania donovani. J Antimicrob Chemo 59 :204–211.
-
17
Dey S, Ouellette M, Lightbody J, Papadopoulou B, Rosen BP, 1996. An ATP-dependent AsIII-glutathione transport system in membrane vesicles of Leishmania tarentolae. Proc Natl Acad Sci USA 93 :2192–2197.
-
18
Gourbal B, Sonuc N, Bhattacharjee H, Legare D, Sundar S, Ouellette M, Rosen BP, Mukhopadhyay R, 2004. Drug uptake and modulation of drug resistance in Leishmania by an aquaglyceroporin. J Biol Chem 279 :31010–31017.
-
19
Marquis N, Gourbal B, Rosen BP, Mukhopadhyay R, Ouellette M, 2005. Modulation in aquaglyceroporin AQP1 gene transcript levels in drug-resistant Leishmania. Mol Microbiol 57 :1690–1699.
-
20
Borgnia M, Nielsen S, Engel A, Aqre P, 1999. Cellular and molecular biology of aquaporin water channels. Annu Rev Biochem 68 :425–458.
-
21
Sanders OI, Rensing C, Kuroda M, Mitra B, Rosen BP, 1997. Antimonite is accumulated by the glycerol facilitator GlpF in Escherichia coli. J Bacteriol 179 :3365–3367.
-
22
Wysocki R, Chery CC, Wawrzycka D, Hulle MV, Cornelis R, Thevelein JM, Tamas MJ, 2001. The glycerol channel Fps1p mediates the uptake of arsenite and antimonite in Saccharomyces cerevisiae. Mol Microbiol 40 :1391–1401.
-
23
Bellofato V, Cross GA, 1989. Expression of a bacterial gene in a trypanosomatid protozoan. Science 244 :1167–1169.
-
24
Brouchu C, Wang J, Roy G, Messier N, Wang XY, Saravia NG, Ouellette M, 2003. Antimony uptake systems in the protozoan parasite Leishania and accumulation differences in antimony-resistant parasites. Antimicrob Agents Chemother 47 :3073–3079.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 323 | 243 | 111 |
| Full Text Views | 366 | 12 | 0 |
| PDF Downloads | 133 | 15 | 0 |
Role of Aquaglyceroporin (AQP1) Gene and Drug Uptake in Antimony-resistant Clinical Isolates of Leishmania donovani
Mahendra Maharjan
Mahendra Maharjan
School of Life Sciences, Jawaharlal Nehru University, New Delhi, India; Department of Pharmacology, Institute of Postgraduate Medical Education and Research, Kolkata, India
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Sushma Singh
Sushma Singh
School of Life Sciences, Jawaharlal Nehru University, New Delhi, India; Department of Pharmacology, Institute of Postgraduate Medical Education and Research, Kolkata, India
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Mitali Chatterjee
Mitali Chatterjee
School of Life Sciences, Jawaharlal Nehru University, New Delhi, India; Department of Pharmacology, Institute of Postgraduate Medical Education and Research, Kolkata, India
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Rentala Madhubala
Rentala Madhubala
School of Life Sciences, Jawaharlal Nehru University, New Delhi, India; Department of Pharmacology, Institute of Postgraduate Medical Education and Research, Kolkata, India
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Antimonial-containing drugs are the first line of treatment against Leishmaniasis. Resistance to antimonials in Leishmania is proposed to be due to reduced uptake of trivalent antimony (SbIII) through the aquaglyceroporin (AQP1). We investigated the uptake of SbIII and involvement of aquaglyceroporin in developing antimony resistance phenotype in Leishmania donovani clinical isolates. SbIII accumulation, copy number of AQP1 gene, and transcript levels were compared in antimony-sensitive versus -resistant isolates. Antimony-resistant field isolates showed reduced uptake of SbIII. The copy number of AQP1 gene showed higher copy number in the antimony-resistant isolates when compared with the sensitive isolates and did not correlate to the reduced uptake of SbIII. Downregulation of AQP1 RNA levels was not consistently found in the antimony-resistant isolates. Our studies indicate that while downregulation of AQP1 may be one of the mechanisms of antimony resistance, it is however not an invariable feature.
Author Notes
- INTRODUCTION
- MATERIALS AND METHODS
- Parasite and culture condition.
- Chemosensitivity profiles of SAG-S and SAG-R promasti-gotes to potassium antimony tartrate (SbIII).
- Uptake assay.
- Cloning of full-length aquaglyceroporin (AQP1) gene from antimony-sensitive and antimony-resistant L. donovani strains.
- Nucleic acid isolation and hybridization analysis.
- RESULTS
- Susceptibility of Leishmania isolates to potassium antimony tartrate (SbIII).
- Comparison of transport of potassium antimony tartrate (SbIII) in SAG-R versus SAG-S field isolates.
- Southern blot analysis of the aquaglyceroporin (AQP1) gene in SAG-S and SAG-R field isolates.
- Comparison of aquaglyceroporin gene expression in SAG-S versus SAG-R field isolates.
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Guerin PJ, Olliaro P, Sundar S, Boelaert M, Croft SL, Desjeux P, Wasunna MK, Bryceson ADM, 2002. Visceral leishmaniasis: current status of control, diagnosis, and treatment, and a proposed research and development agenda. Lancet Infect Dis 2 :494–501.
-
2
Roberts WL, Berman JD, Rainey PM, 1995. In vitro antileishmanial properties of tri- and pentavalent antimonial preparations. Antimicrob Agents Chemother 39 :1234–1239.
-
3
Shaked-Mishan P, Ulrich N, Ephros M, Zilberstein D, 2001. Novel intracellular SbV reducing activity correlates with antimony susceptibility in Leishmania donovani. J Biol Chem 276 :3971–3976.
-
4
Singh AK, Liu HY, Lee ST, 1994. Atomic absorption spectro-photometric measurement of intracellular arsenite in arsenite-resistant Leishmania. Mol Biochem Parasitol 66 :161–164.
-
5
Denton H, McGregor JC, Coombs GH, 2004. Reduction of anti-leishmanial pentavalent antimonial drugs by a parasite-specific thiol-dependent reductase, TDR1. Biochem J 381 :405–412.
-
6
Zhou Y, Messier N, Ouellette M, Rosen BP, Mukhopadhyay R, 2004. Leishmania major LmACR2 is a pentavalent antimony reductase that confers sensitivity to the drug pentostam. J Biol Chem 279 :37445–37451.
-
7
Sereno D, Cavaleyra M, Zemzoumi K, Maquaire S, Ouaissi A, Lemesre JL, 1998. Axenically grown amastigotes of Leishmania infantum used as an in vitro model to investigate the pentavalent antimony mode of action. Antimicrob Agents Chemother 42 :3097–3102.
-
8
Sundar S, More DK, Singh MK, Singh VP, Sharma S, Makharia A, Kumar PCK, Murray W, 2000. Failure of pentavalent antimony in visceral leishmaniasis in India: report from the centre of the Indian epidemic. Clin Infect Dis 31 :1104–1107.
-
9
Lira R, Sundar S, Makharia A, Kenney R, Gam A, Saraiva E, Sacks D, 1999. Evidence that the high incidence of treatment failures in Indian kala-azar is due to the emergence of antimony-resistant strains of Leishmania donovani. J Infect Dis 180 :564–567.
-
10
Sundar S, 2001. Drug resistant in Indian Visceral leishmaniasis. Trop Med Int Health 6 :849–854.
-
11
Mukhopadhyay R, Dey S, Xu N, Gage D, Lightbody J, Ouellette M, Rosen BP, 1996. Trypanothione overproduction and resistance to antimonials and arsenicals in Leishmania. Proc Natl Acad Sci USA 93 :10383–10387.
-
12
Haimeur A, Guimond C, Pilote S, Mukhopadhyay R, Rosen BP, Poulin R, Ouellette M, 1999. Elevated levels of polyamines and trypanothione resulting from overexpression of the ornithine decarboxylase gene in arsenite-resistant Leishmania. Mol Microbiol 34 :726–735.
-
13
Haimeur A, Brochu C, Genest PA, Papadopoulou B, Ouellette M, 2000. Amplification of the ABC transporter gene PGPA and increased trypanothione levels in potassium antimonyl tartrate (SbIII) resistant Leishmania tarentolae. Mol Biochem Parasitol 108 :131–135.
-
14
Legare D, Papadopoulou B, Roy G, Mukhopadhyay R, Haimeur A, Dey S, Grondin K, Brochu C, Rosen BP, Ouellette M, 1997. Efflux systems and increased trypanothione levels in arsenite-resistant Leishmania. Exp Parasitol 87 :275–282.
-
15
Legare D, Richard D, Mukhopadhyay R, Stierhof YD, Rosen BP, Haimeur A, Papadopoulou B, Ouellette M, 2001. The Leishmania ABC protein PGPA is an intracellular metal-thiol transporter ATPase. J Biol Chem 276 :26301–26307.
-
16
Mukherjee A, Padmanabhan PK, Singh S, Roy G, Girard I, Chatterjee M, Ouellette M, Madhubala R, 2007. Role of ABC transporter MRPA, γ-glutamylcysteine synthetase and ornithine decarboxylase in natural antimony-resistant isolates of Leishmania donovani. J Antimicrob Chemo 59 :204–211.
-
17
Dey S, Ouellette M, Lightbody J, Papadopoulou B, Rosen BP, 1996. An ATP-dependent AsIII-glutathione transport system in membrane vesicles of Leishmania tarentolae. Proc Natl Acad Sci USA 93 :2192–2197.
-
18
Gourbal B, Sonuc N, Bhattacharjee H, Legare D, Sundar S, Ouellette M, Rosen BP, Mukhopadhyay R, 2004. Drug uptake and modulation of drug resistance in Leishmania by an aquaglyceroporin. J Biol Chem 279 :31010–31017.
-
19
Marquis N, Gourbal B, Rosen BP, Mukhopadhyay R, Ouellette M, 2005. Modulation in aquaglyceroporin AQP1 gene transcript levels in drug-resistant Leishmania. Mol Microbiol 57 :1690–1699.
-
20
Borgnia M, Nielsen S, Engel A, Aqre P, 1999. Cellular and molecular biology of aquaporin water channels. Annu Rev Biochem 68 :425–458.
-
21
Sanders OI, Rensing C, Kuroda M, Mitra B, Rosen BP, 1997. Antimonite is accumulated by the glycerol facilitator GlpF in Escherichia coli. J Bacteriol 179 :3365–3367.
-
22
Wysocki R, Chery CC, Wawrzycka D, Hulle MV, Cornelis R, Thevelein JM, Tamas MJ, 2001. The glycerol channel Fps1p mediates the uptake of arsenite and antimonite in Saccharomyces cerevisiae. Mol Microbiol 40 :1391–1401.
-
23
Bellofato V, Cross GA, 1989. Expression of a bacterial gene in a trypanosomatid protozoan. Science 244 :1167–1169.
-
24
Brouchu C, Wang J, Roy G, Messier N, Wang XY, Saravia NG, Ouellette M, 2003. Antimony uptake systems in the protozoan parasite Leishania and accumulation differences in antimony-resistant parasites. Antimicrob Agents Chemother 47 :3073–3079.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 323 | 243 | 111 |
| Full Text Views | 366 | 12 | 0 |
| PDF Downloads | 133 | 15 | 0 |