Abstract
Cutaneous leishmaniasis (CL) is a major disease in many parts of the world. Since no vaccine has been developed, treatment is the best way to control it. In most areas, antimonial resistance whose mechanisms have not been completely understood has been reported. The main aim of this study is gene expression assessing of J-binging protein 1 and J-binding protein 2 in clinical Leishmania major isolates. The patients with CL from central and north Iran were considered for this study. The samples were transferred in RNAlater solution and stored in − 20 °C. RNA extraction and cDNA synthesis were performed. The gene expression analysis was done with SYBR Green real-time PCR using ∆∆CT. Written informed consent forms were filled out by patients, and then, information forms were filled out based on the Helsinki Declaration. Statistical analysis was done with SPSS (16.0; SPSS Inc, Chicago) using independent t test, Shapiro–Wilk, and Pearson’s and Spearman’s rank correlation coefficients. P ≤ 0.05 was considered significant. The gene expression of JBP1 and JBP2 had no relation with sex and age. The JBP1 gene expression was high in sensitive isolates obtained from north of the country. The JBP2 gene expression was significant in sensitive and no response-antimonial isolates from the north, but no significant differences were detected in sensitive and resistant isolates from central Iran. Differential gene expression of JBP1 and JBP2 in various clinical resistances isolates in different geographical areas shows multifactorial ways of developing resistance in different isolates.
Similar content being viewed by others
Abbreviations
- CL:
-
Cutaneous leishmaniasis (CL)
- ACL:
-
Anthroponotic cutaneous leishmaniasis
- ZCL:
-
Zoonotic cutaneous leishmaniasis
- JBP:
-
J-binding protein
- RNAP II:
-
RNA polymerase II
- ITS:
-
Internal transcribed spacer
- GAPDH:
-
Glyceraldehyde-3-phosphate dehydrogenase
References
Adaui V, Schnorbusch K, Zimic M, Gutiérrez A, Decuypere S, Vanaerschot M et al (2011) Comparison of gene expression patterns among Leishmania braziliensis clinical isolates showing a different in vitro susceptibility to pentavalent antimony. Parasitology 138:183–193
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W et al (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
Altschul SF, Wootton JC, Gertz EM, Agarwala R, Morgulis A, Schäffer AA et al (2005) Protein database searches using compositionally adjusted substitution matrices. FEBS J 272:5101–5109
Borst P, Sabatini R (2008) Base J: discovery, biosynthesis, and possible functions. Annu Rev Microbiol 62:235–251
Brochu C, Haimeur A, Ouellette M (2004) The heat shock protein HSP70 and heat shock cognate protein HSC70 contribute to antimony tolerance in the protozoan parasite Leishmania. Cell Stress Chaperones 9:294–303
Campbell DA, Thomas S, Sturm N (2003) Transcription in kinetoplastid protozoa: why be normal? Microbes Infect 5:1231–1240
Choudhury K, Zander D, Kube M, Reinhardt R, Clos J (2008) Identification of a Leishmania infantum gene mediating resistance to miltefosine and SbIII. Int J Parasitol 38:1411–1423
Clayton CE (2002) Life without transcriptional control? from fly to man and back again. EMBO J 21:1881–1888
Cliffe LJ, Kieft R, Southern T, Birkeland SR, Marshall M, Sweeney K et al (2009) JBP1 and JBP2 are two distinct thymidine hydroxylases involved in J biosynthesis in genomic DNA of African trypanosomes. Nucleic Acids Res 37:1452–1462
Decuypere S, Rijal S, Yardley V, De Doncker S, Laurent T, Khanal B et al (2005) Gene expression analysis of the mechanism of natural Sb (V) resistance in Leishmania donovani isolates from Nepal. Antimicrob Agents Chemother 49:4616–4621
DiPaolo C, Kieft R, Cross M, Sabatini R (2005) Regulation of trypanosome DNA glycosylation by a SWI2/SNF2-like protein. Mol Cell 17:441–451
Downing T, Imamura H, Decuypere S, Clark TG, Coombs GH, Cotton JA et al (2011) Whole genome sequencing of multiple Leishmania donovani clinical isolates provides insights into population structure and mechanisms of drug resistance. Genome Res 21:2143–2156
Ekanayake DK, Sabatini R (2011) Epigenetic regulation of polymerase II transcription initiation in Trypanosoma cruzi: modulation of nucleosome abundance, histone modification, and polymerase occupancy by O-linked thymine DNA glucosylation. Eukaryot Cell 10:1465–1472
Eslami G, Salehi R (2014) Genetic variation in RPOIILS gene encoding RNA polymerase II largest subunit from Leishmania major. Mol Biol Rep 41:2585–2589
Eslami G, Frikha F, Salehi R, Khamesipour A, Hejazi H, Nilforoushzadeh MA (2011) Cloning, expression and dynamic simulation of TRYP6 from Leishmania major (MRHO/IR/75/ER). Mol Biol Rep 38:3765–3776
Eslami G, Salehi R, Khosravi S, Doudi M (2012) Genetic analysis of clinical isolates of Leishmania major from Isfahan, Iran. J Vector Borne Dis 49:168–174
Eslami G, Hajimohammadi B, Jafari AA, Mirzaei F, Gholamrezai M, Anvari H et al (2014) Molecular identification of Leishmania tropica infections in patients with cutaneous leishmaniasis from an endemic central of Iran. Trop Biomed 31:592–599
Eslami G, Zarchi MV, Moradi A, Hejazi SH, Sohrevardi SM, Vakili M et al (2016) Aquaglyceroporin1 gene expression in antimony resistance and susceptible Leishmania major isolates. J Vector Borne Dis 53:370–374
Genest PA, ter Riet B, Dumas C, Papadopoulou B, van Luenen HGAM, Borst P (2005) Formation of linear inverted repeat amplicons following targeting of an essential gene in Leishmania. Nucleic Acids Res 33:1699–1709
Genest PA, Ter Riet B, Cijsouw T, van Luenen HG, Borst P (2007) Telomeric localization of the modified DNA base J in the genome of the protozoan parasite Leishmania. Nucleic Acids Res 35:2116–2124
Gommers-Ampt JH, Van Leeuwen F, de Beer AL, Vliegenthart JF, Dizdaroglu M, Kowalak JA et al (1993) Beta-D-glucosyl-hydroxymethyluracil: a novel modified base present in the DNA of the parasitic protozoan T. brucei. Cell 75:1129–1136
Gourbal B, Sonuc N, Bhattacharjee H, Legare D, Sundar S, Ouellette M et al (2004) Drug uptake and modulation of drug resistance in Leishmania by an aquaglyceroporin. J Biol Chem 279:31010–31017
Guerin PJ, Olliaro P, Sundar S, Boelaert M, Croft SL, Desjeux P et al (2002) Visceral leishmaniasis: current status of control, diagnosis, and treatment, and a proposed research and development agenda. Lancet Infect Dis 2:494–501
Hazelbaker DZ, Buratowski S (2012) Base J: blocking RNA polymerase’s way. Curr Biol 22:R960–R962
Herwaldt BL (1999) Leishmaniasis. Lancet 354:1191–1199
Iyer LM, Tahiliani M, Rao A, Aravind L (2009) Prediction of novel families of enzymes involved in oxidative and other complex modifications of bases in nucleic acids. Cell Cycle 8:1698–1710
Jeddi F, Mary C, Aoun K, Harrat Z, Bouratbine A, Faraut F et al (2014) Heterogeneity of molecular resistance patterns in antimony-resistant field isolates of Leishmania species from the Western Mediterranean area. Antimicrob Agents Chemother 58:4866–4874
Kazemi-Rad E, Mohebali M, Khadem-Erfan MB, Saffari M, Raoofian R, Hajjaran H et al (2013) Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP approach. Exp Parasitol 35:344–349
Kramer S (2012) Developmental regulation of gene expression in the absence of transcriptional control: the case of kinetoplastids. Mol Biochem Parasitol 181:61–72
Kumar D, Singh R, Bhandari V, Kulshrestha A, Negi NS, Salotra P (2012) Biomarkers of antimony resistance: need for expression analysis of multiple genes to distinguish resistance phenotype in clinical isolates of Leishmania donovani. Parasitol Res 111:223–230
Lee TI, Young RA (2000) Transcription of eukaryotic protein-coding genes. Annu Rev Genet 34:77–137
Legare D, Cayer S, Singh AK, Richard D, Papadopoulou B, Ouellette M (2001) ABC proteins of Leishmania. J Bioenerg Biomembr 33:469–474
Mandal S, Maharjan M, Singh S, Chatterjee M, Madhubala R (2010) Assessing aquaglyceroporin gene status and expression profile in antimony-susceptible and-resistant clinical isolates of Leishmania donovani from India. J Antimicrob Chemother 65:496–507
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
Martinez-Calvillo S, Yan S, Nguyen D, Fox M, Stuart KD, Myler PJ (2003) Transcription of Leishmania major Friedlin chromosome 1 initiates in both directions within a single region. Mol Cell 11:1291–1299
Mukherjee A, Padmanabhan PK, Singh S, Roy G, Girard I, Chatterjee M et al (2007) Role of ABC transporter MRPA, γ-glutamylcysteine synthetase and ornithine decarboxylase in natural antimony-resistant isolates of Leishmania donovani. J Antimicrob Chemother 59:204–211
Mukherjee A, Boisvert S, Monte-Neto RL, Coelho AC, Raymond F, Mukhopadhyay R et al (2013) Telomeric gene deletion and intrachromosomal amplification in antimony-resistant Leishmania. Mol Microbiol 88:189–202
Nühs A, Schäfer C, Zander D, Trübe L, Nevado PT, Schmidt S et al (2014) A novel marker, ARM58, confers antimony resistance to Leishmania spp. Int J Parasitol Drugs Drug Resist 4:37–47
Paule MR, White RJ (2000) Survey and summary: transcription by RNA polymerases I and III. Nucleic Acids Res 28:1283–1298
Tahiliani M, Koh KP, Shen Y, Pastor WA, Bandukwala H, Brudno Y et al (2009) Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1. Science 324:930–935
Ubeda JM, Legare D, Raymond F, Ouameur AA, Boisvert S, Rigault P et al (2008) Modulation of gene expression in drug resistant Leishmania is associated with gene amplification, gene deletion and chromosome aneuploidy. Genome Biol 9:R115
van Luenen HG, Farris C, Jan S, Genest PA, Tripathi P, Velds A et al (2012) Glucosylated hydroxymethyluracil, DNA base J, prevents transcriptional readthrough in leishmania. Cell 150:909–921
Vergnes B, Gourbal B, Girard I, Sundar S, Drummelsmith J, Ouellette M (2007) 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 6:88–101
Yaghoobi-Ershadi MR, Jafari R, Hanafi-Bojd AA (2004) A new epidemic focus of zoonotic cutaneous leishmaniasis in central Iran. Ann Saudi Med 24:98–101
Yu Z, Genest PA, ter Riet B, Sweeney K, DiPaolo C, Kieft R et al (2007) The protein that binds to DNA base J in trypanosomatids has features of a thymidine hydroxylase. Nucleic Acids Res 35:2107–2115
Acknowledgements
The authors would like to thank the technical support from Research Center for Food Hygiene and Safety, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. This research was financially supported by Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Author information
Authors and Affiliations
Contributions
GE conceived, designed, and drafted this manuscript. MV performed the statistical analysis. VA obtained the samples. AF drafted the manuscript. SSH and SA performed the laboratory techniques and quality control. ME reviewed the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Ahmadian, S., Eslami, G., Fatahi, A. et al. J-binding protein 1 and J-binding protein 2 expression in clinical Leishmania major no response-antimonial isolates. J Parasit Dis 43, 39–45 (2019). https://doi.org/10.1007/s12639-018-1052-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12639-018-1052-5