Parasites, Hosts and Diseases

  • 제62권2호
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  • Pages.226-237
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  • 2024
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  • 2982-5164(pISSN)
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  • 1738-0006(eISSN)
대한기생충학·열대의학회 (The Korea Society for Parasitology and Tropical Medicine)
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Molecular cloning, identification, transcriptional analysis, and silencing of enolase on the life cycle of Haemaphysalis longicornis (Acari, Ixodidae) tick

  • Md. Samiul Haque (Laboratory of Veterinary Parasitology, College of Veterinary Medicine and Bio-Safety Research Center, Jeonbuk National University) ;
  • Md. Khalesur Rahman (Department of Microbiology, Faculty of Veterinary and Animal Science, Hajee Mohammad Danesh Science and Technology University) ;
  • Mohammad Saiful Islam (Laboratory of Veterinary Parasitology, College of Veterinary Medicine and Bio-Safety Research Center, Jeonbuk National University) ;
  • Myung-Jo You (Laboratory of Veterinary Parasitology, College of Veterinary Medicine and Bio-Safety Research Center, Jeonbuk National University)
  • 투고 : 2023.03.04
  • 심사 : 2023.05.10
  • 발행 : 2024.05.31
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초록

Ticks, blood-sucking ectoparasites, spread diseases to humans and animals. Haemaphysalis longicornis is a significant vector for tick-borne diseases in medical and veterinary contexts. Identifying protective antigens in H. longicornis for an anti-tick vaccine is a key tick control strategy. Enolase, a multifunctional protein, significantly converts D-2-phosphoglycerate and phosphoenolpyruvate in glycolysis and gluconeogenesis in cell cytoplasm. This study cloned a complete open reading frame (ORF) of enolase from the H. longicornis tick and characterized its transcriptional and silencing effect. We amplified the full-length cDNA of the enolase gene using rapid amplification of cDNA ends. The complete cDNA, with an ORF of 1,297 nucleotides, encoded a 432-amino acid polypeptide. Enolase of the Jeju strain H. longicornis exhibited the highest sequence similarity with H. flava (98%), followed by Dermacentor silvarum (82%). The enolase motifs identified included N-terminal and C-terminal regions, magnesium binding sites, and several phosphorylation sites. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that enolase mRNA transcripts were expressed across all developmental stages of ticks and organs such as salivary gland and midgut. RT-PCR showed higher transcript levels in syn-ganglia, suggesting that synganglion nerves influence enolase's role in tick salivary glands. We injected enolase double-stranded RNA into adult unfed female ticks, after which they were subsequently fed with normal unfed males until they spontaneously dropped off. RNA interference significantly (P<0.05) reduced feeding and reproduction, along with abnormalities in eggs (no embryos) and hatching. These findings suggest enolase is a promising target for future tick control strategies.

키워드

과제정보

This research was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry, and Fisheries (IPET) through the Agriculture, Food, and Rural Affairs Convergence Technologies Program for Educating Creative Global Leaders, funded by the Ministry of Agriculture, Food, and Rural Affairs (MAFRA) (grant number: 320005-4).

참고문헌

  1. Chae JB, Cho YS, Cho YK, Kang JG, Shin NS, et al. Epidemiological investigation of tick species from near domestic animal farms and cattle, goat, and wild boar in Korea. Parasites Hosts Dis 2019;57(3):319-324. https://doi.org/10.3347/kjp.2019.57.3.319 
  2. Abbas RZ, Zaman MA, Colwell DD, Gilleard J, Iqbal Z. Acaricide resistance in cattle ticks and approaches to its management: the state of play. Vet Parasitol 2014;203(1-2):6-20. https://doi.org/10.1016/j.vetpar.2014.03.006 
  3. Tirloni L, Islam MS, Kim TK, Diedrich JK, Yates JR, et al. Saliva from nymph and adult females of Haemaphysalis longicornis: a proteomic study. Parasit Vectors 2015;8(1):338. https://doi.org/10.1186/s13071-015-0918-y 
  4. Diaz-Martin V, Manzano-Roman R, Oleaga A, Encinas-Grandes A, Perez-Sanchez R. Cloning and characterization of a plasminogen-binding enolase from the saliva of the argasid tick Ornithodoros moubata. Vet Parasitol 2013;191(3-4):301-314. https://doi.org/10.1016/j.vetpar.2012.09.019 
  5. Wang WX, Li KL, Chen Y, Lai FX, Fu Q. Identification and function analysis of enolase gene NlEno1 from Nilaparvata lugens (Stal) (Hemiptera:Delphacidae). J Insect Sci 2015;15(1):66. https://doi.org/10.1093/jisesa/iev046 
  6. Xu XL, Cheng TY, Yang H. Enolase, a plasminogen receptor isolated from salivary gland transcriptome of the ixodid tick Haemaphysalis flava. Parasitol Res 2016;115(5):1955-1964. https://doi.org/10.1007/s00436-016-4938-0 
  7. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 1998;391(6669):806-811. https://doi.org/10.1038/35888 
  8. Rahman MK, Kim B, You M. Molecular cloning, expression and impact of ribosomal protein S-27 silencing in Haemaphysalis longicornis (Acari: Ixodidae). Exp Parasitol 2020;209:107829. https://doi.org/10.1016/j.exppara.2019.107829 
  9. Diaz-Martin V, Manzano-Roman R, Oleaga A, Encinas-Grandes A, Perez-Sanchez R. Cloning and characterization of a plasminogen-binding enolase from the saliva of the argasid tick Ornithodoros moubata. Vet Parasitol 2013;191(3-4):301-314. https://doi.org/10.1016/j.vetpar.2012.09.019 
  10. Di Tommaso P, Moretti S, Xenarios I, Orobitg M, Montanyola A, et al. T-Coffee: a web server for the multiple sequence alignment of protein and RNA sequences using structural information and homology extension. Nucleic Acids Res 2011;39(suppl):W13-W7. https://doi.org/10.1093/nar/gkr245 
  11. Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, et al. Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 2011;7:539. https://doi.org/10.1038/msb.2011.75 
  12. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 2018;35(6):1547-1549. https://doi.org/10.1093/molbev/msy096 
  13. Rahman MK, Saiful Islam M, You M. Impact of subolesin and cystatin knockdown by RNA interference in adult female Haemaphysalis longicornis (Acari: Ixodidae) on blood engorgement and reproduction. Insects 2018;9(2):39. https://doi.org/10.3390/insects9020039 
  14. de la Fuente J, Kocan KM, Almazan C, Blouin EF. RNA interference for the study and genetic manipulation of ticks. Trends Parasitol 2007;23(9):427-433. https://doi.org/10.1016/j.pt.2007.07.002 
  15. Karim S, Miller NJ, Valenzuela J, Sauer JR, Mather TN. RNAi-mediated gene silencing to assess the role of synaptobrevin and cystatin in tick blood feeding. Biochem Biophys Res Commun 2005;334(4):1336-1342. https://doi.org/10.1016/j.bbrc.2005.07.036 
  16. de la Fuente J, Almazan C, Blouin EF, Naranjo V, Kocan KM. Reduction of tick infections with Anaplasma marginale and A. phagocytophilum by targeting the tick protective antigen subolesin. Parasitol Res 2006;100(1):85-91. https://doi.org/10.1007/s00436-006-0244-6 
  17. Commins SP, James HR, Kelly LA, Pochan SL, Workman LJ, et al. The relevance of tick bites to the production of IgE antibodies to the mammalian oligosaccharide galactose-α-1, 3-galactose. J Allergy Clin Immunol Pract 2011;127(5):1286-1293.e6. https://doi.org/10.1016/j.jaci.2011.02.019 
  18. Kocan KM, Manzano-Roman R, de la Fuente J. Transovarial silencing of the subolesin gene in three-host ixodid tick species after injection of replete females with subolesin dsRNA. Parasitol Res 2007;100(6):1411-1415. https://doi.org/10.1007/s00436-007-0483-1 
  19. Rahman MK, You M. Molecular cloning and transcriptional and functional analysis of glycogen synthase kinase-3β in Haemaphysalis longicornis (Acari, Ixodidae). Parasite 2019;26:39. https://doi.org/10.1051/parasite/2019038 
  20. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001;29(9):e45. https://doi.org/10.1093/nar/29.9.e45 
  21. Pancholi V. Multifunctional α-enolase: its role in diseases. Cell Mol Life Sci 2001;58(7):902-920. https://doi.org/10.1007/pl00000910 
  22. Sigrist CJA, De Castro E, Cerutti L, Cuche BA, Hulo N, et al. New and continuing developments at PROSITE. Nucleic Acids Res 2012;41(database issue):344-347. https://doi.org/10.1093/nar/gks1067 
  23. Cao H, Zhou Y, Chang Y, Zhang X, Li C, et al. Comparative phosphoproteomic analysis of developing maize seeds suggests a pivotal role for enolase in promoting starch synthesis. Plant Sci 2019;289:110243. https://doi.org/10.1016/j.plantsci.2019.110243 
  24. Quintero-Troconis E, Buelvas N, Carrasco-Lopez C, Domingo-Sananes M, Gonzalez-Gonzalez L, et al. Enolase from Trypanosoma cruzi is inhibited by its interaction with metallocarboxypeptidase-1 and a putative acireductone dioxygenase. Biochim Biophys Acta Proteins Proteom 2018;1866(5-6):651-660. https://doi.org/10.1016/j.bbapap.2018.03.003 
  25. Salazar N, de Souza MCL, Biasioli AG, da Silva LB, Barbosa AS. The multifaceted roles of Leptospira enolase. Res Microbiol 2017;168(2):157-164. https://doi.org/10.1016/j.resmic.2016.10.005 
  26. Neelakanta G, Sultana H. Tick saliva and salivary glands: what do we know so far on their role in arthropod blood feeding and pathogen transmission. Front Cell Infect Microbiol 2022:11:816547. https://doi.org/10.3389/fcimb.2021.816547 
  27. Xu XL, Cheng TY, Yang H. Enolase, a plasminogen receptor isolated from salivary gland transcriptome of the ixodid tick Haemaphysalis flava. Parasitol Res 2016;115(5):1955-1964. https://doi.org/10.1007/s00436-016-4938-0 
  28. Avilan L, Gualdron-Lopez M, Quinones W, Gonzalez-Gonzalez L, Hannaert V, et al. Enolase: a key player in the metabolism and a probable virulence factor of trypanosomatid parasites-perspectives for its use as a therapeutic target. Enzyme Res 2011;2011:932549. https://doi.org/10.4061/2011/932549 
  29. Piast M, Kustrzeba-Wojcicka I, Matusiewicz M, Banas T. Molecular evolution of enolase. Acta Biochim Pol 2005;52(2):507-513. 
  30. Haque A, Ray SK, Cox A, Banik NL. Neuron specific enolase: a promising therapeutic target in acute spinal cord injury. Metab Brain Dis 2016;31(3):487-495. https://doi.org/10.1007/s11011-016-9801-6 
  31. de la Torre-Escudero E, Manzano-Roman R, Perez-Sanchez R, Siles-Lucas M, Oleaga A. Cloning and characterization of a plasminogen-binding surface-associated enolase from Schistosoma bovis. Vet Parasitol 2010;173(1-2):76-84. https://doi.org/10.1016/j.vetpar.2010.06.011 
  32. Diaz-Ramos A, Roig-Borrellas A, Garcia-Melero A, Lopez-Alemany R. α-Enolase, a multifunctional protein: its role on pathophysiological situations. J Biomed Biotechnol 2012;2012. https://doi.org/10.1155/2012/156795 
  33. Imamura S, Junior IdSV, Sugino M, Ohashi K, Onuma M. A serine protease inhibitor (serpin) from Haemaphysalis longicornis as an anti-tick vaccine. Vaccine 2005;23(10):1301-1311. https://doi.org/10.1016/j.vaccine.2004.08.041 
  34. Ren S, Zhang B, Xue X, Wang X, Zhao H, et al. Salivary gland proteome analysis of developing adult female Haemaphysalis longicornis ticks: molecular motor and TCA cycle-related proteins play an important role throughout development. Parasit Vectors 2019;12(1):1-16. https://doi.org/10.1186/s13071-019-3864-2 
  35. Liu L, Yan F, Zhang L, Wu ZF, Duan DY, Cheng TY. Protein profiling of hemolymph in Haemaphysalis flava ticks. Parasit Vectors 2022;15(1):1-12. https://doi.org/10.1186/s13071-022-05287-7 
  36. Xu XL, Cheng TY, Yang H, Yan F, Yang Y. De novo sequencing, assembly and analysis of salivary gland transcriptome of Haemaphysalis flava and identification of sialoprotein genes. Infect Genet Evol 2015;32:135-142. https://doi.org/10.1016/j.meegid.2015.03.010