An OTU Deubiquitinating Enzyme in Eimeria Tenella Interacts with Eimeria Tenella Virus RDRP

Overview

Journal Parasit Vectors

Publisher Biomed Central

Specialties Parasitology
Tropical Medicine

Date 2018 Feb 2

PMID 29386062

Citations 4

Authors

Pu Wang

Jianhua Li

Pengtao Gong

Weirong Wang

Yongxing Ai

Xichen Zhang

Affiliations

Soon will be listed here.

Abstract

Background: Chicken coccidiosis, a disease caused by seven species of Eimeria (Apicomplexa: Coccidia), inflicts severe economic losses on the poultry industry. Eimeria tenella is the one of the most virulent species pathogenic to chickens. Many parasitic protozoans are parasitised by double-stranded (ds) RNA viruses, and the influence of protozoan viruses on parasitic protozoans has been extensively reported. E. tenella RNA virus 1 (Etv) was identified in E. tenella, and the complete genome sequence of Etv was analysed. Here, we screened Etv-RNA-dependent RNA polymerase (RDRP)-interacting host protein E. tenella ovarian tumour (OTU) protein-like cysteine protease (Et-OTU) using a yeast two-hybrid system with pGBKT7-RDRP plasmid serving as bait. A previous study demonstrated that Et-OTU could regulate the telomerase activity of E. tenella, indicating that Et-OTU affects E. tenella proliferation. However, whether Etv-RDRP affects the molecular biological characteristics of E. tenella by interacting with OTU remains unclear.

Results: We obtained seven positive clones from the initial screen, and six of the seven preys were identified as false-positives. Finally, we identified an RDRP-associated protein predicted to be an E. tenella OTU protein. A α-galactosidase assay showed that the bait vector did not activate the GAL4 reporter gene, indicating no autoactivation activity from the RDRP bait fusion. Pull-down and co-immunoprecipitation assays verified the interaction between Et-OTU and Etv-RDRP both intracellularly and extracellularly. Additionally, Et-OTU was able to deconjugate K48- and K6-linked di-ubiquitin (di-Ub) chains in vitro but not K63-, K11-, K29-, or K33-linked di-Ub chains. The C239A and H351A mutations eliminated the deubiquitinase (DUB) activity of Et-OTU, whereas the D236A mutation did not. Additionally, when combined with RDRP, the DUB activity of Et-OTU towards K48- and K6-linked chains was significantly enhanced.

Conclusion: Etv-RDRP interacts with Et-OTU both intracellularly and extracellularly. Etv-RDRP enhances the hydrolysis of Et-OTU to K6- or K48-linked ubiquitin chains. This study lays the foundation for further research on the relationship between E. tenella and Etv.

Citing Articles

A potential role for Giardia chaperone protein GdDnaJ in regulating Giardia proliferation and Giardiavirus replication.

Zhang H, Zhao C, Zhang X, Li J, Gong P, Wang X Parasit Vectors. 2023; 16(1):168.

PMID: 37226181 PMC: 10210397. DOI: 10.1186/s13071-023-05787-0.

Characterisation of the OTU domain deubiquitinase complement of .

Wilde M, Ruparel U, Klemm T, Lee V, Calleja D, Komander D Life Sci Alliance. 2023; 6(6).

PMID: 36958824 PMC: 10038098. DOI: 10.26508/lsa.202201710.

The emerging role of Deubiquitinases (DUBs) in parasites: A foresight review.

Kumar P, Kumar P, Mandal D, Velayutham R Front Cell Infect Microbiol. 2022; 12:985178.

PMID: 36237424 PMC: 9552668. DOI: 10.3389/fcimb.2022.985178.

Identification and Protective Efficacy of Rhoptry Kinase Family Protein 17.

Liu X, Mu B, Zheng W, Meng Y, Yu L, Gao W Animals (Basel). 2022; 12(5).

PMID: 35268126 PMC: 8908856. DOI: 10.3390/ani12050556.

References

Mevissen T, Hospenthal M, Geurink P, Elliott P, Akutsu M, Arnaudo N . OTU deubiquitinases reveal mechanisms of linkage specificity and enable ubiquitin chain restriction analysis. Cell. 2013; 154(1):169-84. PMC: 3705208. DOI: 10.1016/j.cell.2013.05.046. View

Wu B, Zhang X, Gong P, Li M, Ding H, Xin C . Eimeria tenella: a novel dsRNA virus in E. tenella and its complete genome sequence analysis. Virus Genes. 2016; 52(2):244-52. DOI: 10.1007/s11262-016-1295-0. View

Yeh J, Wang C . Telomeres and Telomerase in Cardiovascular Diseases. Genes (Basel). 2016; 7(9). PMC: 5042389. DOI: 10.3390/genes7090058. View

Amerik A, Hochstrasser M . Mechanism and function of deubiquitinating enzymes. Biochim Biophys Acta. 2004; 1695(1-3):189-207. DOI: 10.1016/j.bbamcr.2004.10.003. View

Revets H, Dekegel D, Deleersnijder W, De Jonckheere J, Peeters J, Leysen E . Identification of virus-like particles in Eimeria stiedae. Mol Biochem Parasitol. 1989; 36(3):209-15. DOI: 10.1016/0166-6851(89)90168-0. View

Durcan T, Fon E . USP8 and PARK2/parkin-mediated mitophagy. Autophagy. 2015; 11(2):428-9. PMC: 4502724. DOI: 10.1080/15548627.2015.1009794. View

Durcan T, Tang M, Perusse J, Dashti E, Aguileta M, McLelland G . USP8 regulates mitophagy by removing K6-linked ubiquitin conjugates from parkin. EMBO J. 2014; 33(21):2473-91. PMC: 4283406. DOI: 10.15252/embj.201489729. View

Tenno T, Fujiwara K, Tochio H, Iwai K, Hayato Morita E, Hayashi H . Structural basis for distinct roles of Lys63- and Lys48-linked polyubiquitin chains. Genes Cells. 2004; 9(10):865-75. DOI: 10.1111/j.1365-2443.2004.00780.x. View

Komander D . The emerging complexity of protein ubiquitination. Biochem Soc Trans. 2009; 37(Pt 5):937-53. DOI: 10.1042/BST0370937. View

10.

Wang A, Wang C . Discovery of a specific double-stranded RNA virus in Giardia lamblia. Mol Biochem Parasitol. 1986; 21(3):269-76. DOI: 10.1016/0166-6851(86)90132-5. View

11.

Crosas B . Deubiquitinating enzyme inhibitors and their potential in cancer therapy. Curr Cancer Drug Targets. 2014; 14(6):506-16. DOI: 10.2174/1568009614666140725090620. View

12.

Wang P, Wang W, Yang J, Ai Y, Gong P, Zhang X . A novel telomerase-interacting OTU protein of Eimeria tenella and its telomerase-regulating activity. Acta Biochim Biophys Sin (Shanghai). 2017; 49(8):744-745. DOI: 10.1093/abbs/gmx057. View

13.

Hanpude P, Bhattacharya S, Dey A, Maiti T . Deubiquitinating enzymes in cellular signaling and disease regulation. IUBMB Life. 2015; 67(7):544-55. DOI: 10.1002/iub.1402. View

14.

Spivak I, Mikhelson V, Spivak D . [Telomere length, telomerase activity, stress and aging]. Adv Gerontol. 2017; 28(3):441-448. View

15.

Yin H, Sumners L, Dalloul R, Miska K, Fetterer R, Jenkins M . Changes in expression of an antimicrobial peptide, digestive enzymes, and nutrient transporters in the intestine of E. praecox-infected chickens. Poult Sci. 2015; 94(7):1521-6. DOI: 10.3382/ps/pev133. View

16.

Dhara A, Sinai A . A Cell Cycle-Regulated Toxoplasma Deubiquitinase, TgOTUD3A, Targets Polyubiquitins with Specific Lysine Linkages. mSphere. 2016; 1(3). PMC: 4917281. DOI: 10.1128/mSphere.00085-16. View

17.

Butcher S, Grimes J, Makeyev E, Bamford D, Stuart D . A mechanism for initiating RNA-dependent RNA polymerization. Nature. 2001; 410(6825):235-40. DOI: 10.1038/35065653. View

18.

Lim K, Ramakrishna S, Baek K . Molecular mechanisms and functions of cytokine-inducible deubiquitinating enzymes. Cytokine Growth Factor Rev. 2013; 24(5):427-31. DOI: 10.1016/j.cytogfr.2013.05.007. View

19.

Xie J, Havens W, Lin Y, Suzuki N, Ghabrial S . Reprint of "The victorivirus Helminthosporium victoriae virus 190S is the primary cause of disease/hypovirulence in its natural host and a heterologous host". Virus Res. 2016; 219:100-107. DOI: 10.1016/j.virusres.2016.05.010. View

20.

York A, Hutchinson E, Fodor E . Interactome analysis of the influenza A virus transcription/replication machinery identifies protein phosphatase 6 as a cellular factor required for efficient virus replication. J Virol. 2014; 88(22):13284-99. PMC: 4249064. DOI: 10.1128/JVI.01813-14. View