An Eimeria acervulina OTU protease exhibits linkage-specific deubiquitinase activity
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Ubiquitination is an important post-translational modification process that regulates many cellular processes. Proteins can be modified at single or multiple lysine residues by a single ubiquitin protein or by ubiquitin oligomers. It is important to note that the type of ubiquitin chains determines the functional outcome of the modification. Ubiquitin or ubiquitin chains can be removed by deubiquitinases (DUBs). In our previous study, the Eimeria tenella ovarian tumour (Et-OTU) DUB was shown to regulate the telomerase activity of E. tenella and affect E. tenella proliferation. The amino acid sequences of Et-OTU (GenBank: XP_013229759.1) and Eimeria acervulina (E. acervulina) ovarian tumour (Ea-OTUD3) DUB (XP_013250378.1) are 74% identical. Although Et-OTU may regulate E. tenella telomerase activity, whether Ea-OTUD3 affects E. acervulina growth and reproduction remains unclear. We show here that Ea-OTUD3 belongs to the OTU domain class of cysteine protease deubiquitinating enzymes. Ea-OTUD3 is highly linkage-specific, cleaving K48 (Lys48)-, K63-, and K6-linked diubiquitin but not K29-, K33-, and K11-linked diubiquitin. The precise linkage preference of Ea-OTUD3 among these three nonlinear diubiquitin chains is K6 > K48 > K63. Recombinant Ea-OTUD3, but not its catalytic-site mutant Ea-OTUD3 (C247A), exhibits activity against diubiquitin. Ea-OTUD3 removes ubiquitin from the K48-, but to a lesser extent from the K63-linked ubiquitinated E. acervulina proteins of the modified target protein, thereby exhibiting the characteristics of deubiquitinase. This study reveals that the Ea-OTUD3 is a novel functional deubiquitinating enzyme. Furthermore, the Ea-OTUD3 protein may regulate the stability of some K48-linked ubiquitinated E. acervulina proteins.
KeywordsEimeria acervulina Deubiquitinase OTU Ubiquitin
This work was supported by grant from Jilin province science and technology development plan item (no. 20170204036NY) and from the National Natural Science Foundation (NSFC) of China (nos. 31272550, 31672288, and 30970322).
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Conflict of interest
The authors declare that they have no conflicts of interest.
- Datta G, Hossain ME, Asad M, Rathore S, Mohmmed A. (2017). Plasmodium falciparum OTU-like cysteine protease (PfOTU) is essential for apicoplast homeostasis and associates with non-canonical role of Atg8. Cell Microbiol 19(9)Google Scholar
- Dhara A, Sinai AP (2016) A cell cycle-regulated toxoplasma deubiquitinase, TgOTUD3A, targets polyubiquitins with specific lysine linkages. mSphere 1(3):e00085–16Google Scholar
- Dhara A, de Paula Baptista R, Kissinger JC, Snow EC, Sinai AP. (2017). Ablation of an ovarian tumor family deubiquitinase exposes the underlying regulation governing the plasticity of cell cycle progression in Toxoplasma gondii. MBio 8(6): e01846–17Google Scholar
- Groves MR, Schroer CFE, Middleton AJ, Lunev S, Danda N, Ali AM, Marrink SJ, Williams C (2017) Structural insights into K48-linked ubiquitin chain formation by the Pex4p-Pex22p complex. Biochem Biophys Res Commun 496(2):562–567Google Scholar
- Natalia D, MagnaniLaura A, DadaJacob I (2018) Ubiquitin-proteasome signaling in lung injury. Transl Res S1931-5244(18):30057–30054Google Scholar
- Rubtsova MP, Vasilkova DP, Malyavko AN, Naraikina YV, Zvereva MI, Dontsova OA (2012) Telomere lengthening and other functions of telomerase. Acta Nat 4(2):44–61Google Scholar
- Shanmugham A, Ovaa H (2008) DUBs and disease activity assays for inhibitor development. Curr Opin Drug Discov Devel 11(5):688–696Google Scholar
- Staszczak M (2017) Ubiquitin-proteasome pathway as a target for therapeutic strategies. Postepy Biochem 63(4):287–303Google Scholar
- Yu X, Robinson JF, Sidhu JS, Hong S, Faustman EM (2010) A system-based comparison of gene expression reveals alterations in oxidative stress, disruption of ubiquitin-proteasome system and altered cell cycle regulation after exposure to cadmium and methylmercury in mouse embryonic fibroblast. Toxicol Sci 114(2):356–377CrossRefGoogle Scholar