Characterization of protein arginine methyltransferase of TgPRMT5 in Toxoplasma gondii

Min Liu, Fen Xiang Li, Chun Yuan Li, Xiao Cong Li, Long Fei Chen, Kun Wu, Pei Liang Yang, Zhi Fa Lai, Ting Kai Liu, William Sullivan, Liwang Cui, Xiao Guang Chen

Research output: Contribution to journalArticle

Abstract

Background: Protein arginine methylation is a prevalent post-translational modification. The protein arginine methyltransferase family (PRMT) is involved in many cellular processes in eukaryotes, including transcriptional regulation, epigenetic regulation, RNA metabolism, and DNA damage repair. Toxoplasma gondii, an opportunistic protozoan parasite, encodes five conserved PRMTs. PRMT5 is thought to be responsible for substantial PRMT activity in T. gondii; however, it has not yet been characterized. Methods: We tagged the 3′ end of the endogenous TgPRMT5 genomic locus with sequence encoding a 3X hemagglutinin (HA) epitope. IFA and WB were performed to check the expression and subcellular localization of TgPRMT5 in tachyzoites and bradyzoites. In vitro methylation assays were performed to determine whether endogenous TgPRMT5 has arginine methyltransferase activity. Results: IFA and WB results showed that T. gondii PRMT5 (TgPRMT5) was localized in the cytoplasm in the tachyzoite stage; however, it shifts largely to the nuclear compartment in the bradyzoite stage. The in vitro methylation showed that TgPRMT5 has authentic type II PRMT activity and forms monomethylarginines and symmetric dimethylarginines. Conclusions: We determined the expression and cellular localization of TgPRMT5 in tachyzoites and bradyzoites and confirmed its type II PRMT activity. We demonstrated the major changes in expression and cellular localization of TgPRMT5 during the tachyzoite and bradyzoite stages in T. gondii. Our findings suggest that TgPRMT5 protein may be involved in tachyzoite-bradyzoite transformation.

Original languageEnglish (US)
Article number221
JournalParasites and Vectors
Volume12
Issue number1
DOIs
StatePublished - May 8 2019

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Protein-Arginine N-Methyltransferases
Toxoplasma
Methylation
Hemagglutinins
Post Translational Protein Processing
Eukaryota
Epigenomics
DNA Repair
DNA Damage
Arginine
Epitopes

Keywords

  • Bradyzoites
  • Chromatin
  • Epigenetics
  • Histone
  • Methylation
  • Parasites

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

Cite this

Liu, M., Li, F. X., Li, C. Y., Li, X. C., Chen, L. F., Wu, K., ... Chen, X. G. (2019). Characterization of protein arginine methyltransferase of TgPRMT5 in Toxoplasma gondii. Parasites and Vectors, 12(1), [221]. https://doi.org/10.1186/s13071-019-3464-1

Characterization of protein arginine methyltransferase of TgPRMT5 in Toxoplasma gondii. / Liu, Min; Li, Fen Xiang; Li, Chun Yuan; Li, Xiao Cong; Chen, Long Fei; Wu, Kun; Yang, Pei Liang; Lai, Zhi Fa; Liu, Ting Kai; Sullivan, William; Cui, Liwang; Chen, Xiao Guang.

In: Parasites and Vectors, Vol. 12, No. 1, 221, 08.05.2019.

Research output: Contribution to journalArticle

Liu, M, Li, FX, Li, CY, Li, XC, Chen, LF, Wu, K, Yang, PL, Lai, ZF, Liu, TK, Sullivan, W, Cui, L & Chen, XG 2019, 'Characterization of protein arginine methyltransferase of TgPRMT5 in Toxoplasma gondii', Parasites and Vectors, vol. 12, no. 1, 221. https://doi.org/10.1186/s13071-019-3464-1
Liu, Min ; Li, Fen Xiang ; Li, Chun Yuan ; Li, Xiao Cong ; Chen, Long Fei ; Wu, Kun ; Yang, Pei Liang ; Lai, Zhi Fa ; Liu, Ting Kai ; Sullivan, William ; Cui, Liwang ; Chen, Xiao Guang. / Characterization of protein arginine methyltransferase of TgPRMT5 in Toxoplasma gondii. In: Parasites and Vectors. 2019 ; Vol. 12, No. 1.
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AU - Liu, Min

AU - Li, Fen Xiang

AU - Li, Chun Yuan

AU - Li, Xiao Cong

AU - Chen, Long Fei

AU - Wu, Kun

AU - Yang, Pei Liang

AU - Lai, Zhi Fa

AU - Liu, Ting Kai

AU - Sullivan, William

AU - Cui, Liwang

AU - Chen, Xiao Guang

PY - 2019/5/8

Y1 - 2019/5/8

N2 - Background: Protein arginine methylation is a prevalent post-translational modification. The protein arginine methyltransferase family (PRMT) is involved in many cellular processes in eukaryotes, including transcriptional regulation, epigenetic regulation, RNA metabolism, and DNA damage repair. Toxoplasma gondii, an opportunistic protozoan parasite, encodes five conserved PRMTs. PRMT5 is thought to be responsible for substantial PRMT activity in T. gondii; however, it has not yet been characterized. Methods: We tagged the 3′ end of the endogenous TgPRMT5 genomic locus with sequence encoding a 3X hemagglutinin (HA) epitope. IFA and WB were performed to check the expression and subcellular localization of TgPRMT5 in tachyzoites and bradyzoites. In vitro methylation assays were performed to determine whether endogenous TgPRMT5 has arginine methyltransferase activity. Results: IFA and WB results showed that T. gondii PRMT5 (TgPRMT5) was localized in the cytoplasm in the tachyzoite stage; however, it shifts largely to the nuclear compartment in the bradyzoite stage. The in vitro methylation showed that TgPRMT5 has authentic type II PRMT activity and forms monomethylarginines and symmetric dimethylarginines. Conclusions: We determined the expression and cellular localization of TgPRMT5 in tachyzoites and bradyzoites and confirmed its type II PRMT activity. We demonstrated the major changes in expression and cellular localization of TgPRMT5 during the tachyzoite and bradyzoite stages in T. gondii. Our findings suggest that TgPRMT5 protein may be involved in tachyzoite-bradyzoite transformation.

AB - Background: Protein arginine methylation is a prevalent post-translational modification. The protein arginine methyltransferase family (PRMT) is involved in many cellular processes in eukaryotes, including transcriptional regulation, epigenetic regulation, RNA metabolism, and DNA damage repair. Toxoplasma gondii, an opportunistic protozoan parasite, encodes five conserved PRMTs. PRMT5 is thought to be responsible for substantial PRMT activity in T. gondii; however, it has not yet been characterized. Methods: We tagged the 3′ end of the endogenous TgPRMT5 genomic locus with sequence encoding a 3X hemagglutinin (HA) epitope. IFA and WB were performed to check the expression and subcellular localization of TgPRMT5 in tachyzoites and bradyzoites. In vitro methylation assays were performed to determine whether endogenous TgPRMT5 has arginine methyltransferase activity. Results: IFA and WB results showed that T. gondii PRMT5 (TgPRMT5) was localized in the cytoplasm in the tachyzoite stage; however, it shifts largely to the nuclear compartment in the bradyzoite stage. The in vitro methylation showed that TgPRMT5 has authentic type II PRMT activity and forms monomethylarginines and symmetric dimethylarginines. Conclusions: We determined the expression and cellular localization of TgPRMT5 in tachyzoites and bradyzoites and confirmed its type II PRMT activity. We demonstrated the major changes in expression and cellular localization of TgPRMT5 during the tachyzoite and bradyzoite stages in T. gondii. Our findings suggest that TgPRMT5 protein may be involved in tachyzoite-bradyzoite transformation.

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KW - Chromatin

KW - Epigenetics

KW - Histone

KW - Methylation

KW - Parasites

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