Molecular Characterization and Analysis of the Drug Resistance-associated Protein Phosphoglycerate Kinase of

Overview

Journal Parasitology

Specialty Parasitology

Date 2024 Oct 17

PMID 39419817

Authors

Yu Yu

Wenhao Huang

Qiping Zhao

Shunhai Zhu

Hui Dong

Hongyu Han

Affiliations

Soon will be listed here.

Abstract

Coccidiosis is a parasitic disease caused by spp., and the emergence of drug resistance has seriously affected the control of the disease. Using RNA-seq, we previously found that phosphoglycerate kinase of (PGK) was differentially downregulated in diclazuril-resistant (DZR) and maduramicin-resistant (MRR) strains compared with drug-sensitive (DS) strain. In this study, we further analysed the characteristics and functions of PGK to find the possible mechanism of drug resistance of . Quantitative real-time PCR (qRT-PCR) and western blot found that PGK was highly expressed in sporulated oocysts, followed by sporozoites and second-generation merozoites of . Indirect immunofluorescence localization showed that PGK was located mainly in the cytoplasm and on the surface of the parasites. Invasion inhibition assays showed that anti-rPGK antibody significantly inhibited the invasion of parasites. Further studies using qRT-PCR and western blot found that the transcription and translation levels of PGK were downregulated in both resistant (DZR and MRR) strains compared with the DS strain, and the transcription level correlated negatively with the drug concentration. The enzyme activity assay revealed that PGK enzyme activity was decreased in the DZR strain compared with the DS strain. qRT-PCR revealed that the mRNA transcription level of PGK was significantly downregulated in the field DZR strain and salinomycin-resistant strain compared with the DS strain. These results suggested that PGK has other important roles that are separate and distinct from its function in glycolysis, and it might be involved in the development of drug resistance of .

References

Jacot D, Waller R, Soldati-Favre D, MacPherson D, MacRae J . Apicomplexan Energy Metabolism: Carbon Source Promiscuity and the Quiescence Hyperbole. Trends Parasitol. 2015; 32(1):56-70. DOI: 10.1016/j.pt.2015.09.001. View

Mogi T, Kita K . Diversity in mitochondrial metabolic pathways in parasitic protists Plasmodium and Cryptosporidium. Parasitol Int. 2010; 59(3):305-12. DOI: 10.1016/j.parint.2010.04.005. View

Hong S, Shin J, Kang S, Ryu J . Ultrastructural localization of phosphoglycerate kinase in adult Clonorchis sinensis. Parasitol Res. 2003; 90(5):369-71. DOI: 10.1007/s00436-003-0857-y. View

Lou D, Cao Y, Duan H, Tan J, Li B, Zhou Y . Characterization of a Novel Thermostable 7α-Hydroxysteroid Dehydrogenase. Protein Pept Lett. 2024; 31(2):153-160. DOI: 10.2174/0109298665279004231229100320. View

Kazemi-Rad E, Mohebali M, Khadem-Erfan M, Saffari M, Raoofian R, Hajjaran H . Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP approach. Exp Parasitol. 2013; 135(2):344-9. DOI: 10.1016/j.exppara.2013.07.018. View

Miska K, Fetterer R, Barfield R . Analysis of transcripts expressed by Eimeria tenella oocysts using subtractive hybridization methods. J Parasitol. 2005; 90(6):1245-52. DOI: 10.1645/GE-309R. View

Zhou B, Wang H, Wang X, Zhang L, Zhang K, Xue F . Eimeria tenella: effects of diclazuril treatment on microneme genes expression in second-generation merozoites and pathological changes of caeca in parasitized chickens. Exp Parasitol. 2010; 125(3):264-70. DOI: 10.1016/j.exppara.2010.01.022. View

Chen T, Huang B, Zhao Q, Dong H, Zhu S, Zhao Z . Molecular characterization and functional analysis of Eimeria tenella malate dehydrogenase. Parasitol Res. 2018; 117(7):2053-2063. DOI: 10.1007/s00436-018-5875-x. View

Chen S, Wang H, Chen J, Cheng J, Gao J, Chen S . Upregulation of mitochondrial PGK1 by ROS-TBC1D15 pathway promotes neuronal death after oxygen-glucose deprivation/reoxygenation injury. Brain Res. 2023; 1825:148724. DOI: 10.1016/j.brainres.2023.148724. View

10.

Huang W, Zhu S, Chen T, Zhao Q, Dong H, Huang B . Molecular characterization of glyceraldehyde-3-phosphate dehydrogenase from Eimeria tenella. Parasitol Res. 2022; 121(6):1749-1760. DOI: 10.1007/s00436-022-07508-5. View

11.

Birnbaum J, Scharf S, Schmidt S, Jonscher E, Hoeijmakers W, Flemming S . A Kelch13-defined endocytosis pathway mediates artemisinin resistance in malaria parasites. Science. 2020; 367(6473):51-59. DOI: 10.1126/science.aax4735. View

12.

Echaniz-Laguna A, Nadjar Y, Behin A, Biancalana V, Piraud M, Malfatti E . Phosphoglycerate kinase deficiency: A nationwide multicenter retrospective study. J Inherit Metab Dis. 2019; 42(5):803-808. DOI: 10.1002/jimd.12087. View

13.

Hong Y, Huang L, Yang J, Cao X, Han Q, Zhang M . Cloning, expression and enzymatic characterization of 3-phosphoglycerate kinase from Schistosoma japonicum. Exp Parasitol. 2015; 159:37-45. DOI: 10.1016/j.exppara.2015.08.016. View

14.

Banks R, Blake C, Evans P, Haser R, Rice D, Hardy G . Sequence, structure and activity of phosphoglycerate kinase: a possible hinge-bending enzyme. Nature. 1979; 279(5716):773-7. DOI: 10.1038/279773a0. View

15.

Adhikari A, Biswas S, Mukherjee A, Das S, Adak S . PAS domain-containing phosphoglycerate kinase deficiency in results in increased autophagosome formation and cell death. Biochem J. 2019; 476(8):1303-1321. DOI: 10.1042/BCJ20190041. View

16.

Gelinas A, Sudan S, Patterson R, Li J, Huyben D, Barta J . Growth performance, organs weight, intestinal histomorphology, and oocyst shedding in broiler chickens offered novel single strain Bacillus subtilis isolated from camel dung and challenged with Eimeria. Poult Sci. 2024; 103(4):103519. PMC: 10877951. DOI: 10.1016/j.psj.2024.103519. View

17.

Lee H, Cho D, Ahmad I, Patel H, Kim M, Jung J . Mining of a novel esterase (est3S) gene from a cow rumen metagenomic library with organosphosphorus insecticides degrading capability: Catalytic insights by site directed mutations, docking, and molecular dynamic simulations. Int J Biol Macromol. 2021; 190:441-455. DOI: 10.1016/j.ijbiomac.2021.08.224. View

18.

Danne J, Gornik S, MacRae J, McConville M, Waller R . Alveolate mitochondrial metabolic evolution: dinoflagellates force reassessment of the role of parasitism as a driver of change in apicomplexans. Mol Biol Evol. 2012; 30(1):123-39. DOI: 10.1093/molbev/mss205. View

19.

Burgess R . Elution of proteins from gels. Methods Enzymol. 2009; 463:565-72. DOI: 10.1016/S0076-6879(09)63032-9. View

20.

Taylor B, Zhulin I . PAS domains: internal sensors of oxygen, redox potential, and light. Microbiol Mol Biol Rev. 1999; 63(2):479-506. PMC: 98974. DOI: 10.1128/MMBR.63.2.479-506.1999. View