Oncotarget

Research Papers:

Toxoplasma gondii Clp family protein: TgClpB1 plays a crucial role in thermotolerance

Shinuo Cao, Nali Du, Heming Chen, Yu Pang, Zhaoxia Zhang, Jun Zheng and Honglin Jia _

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Oncotarget. 2017; 8:86117-86129. https://doi.org/10.18632/oncotarget.20989

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Abstract

Shinuo Cao1,*, Nali Du1,*, Heming Chen1, Yu Pang1, Zhaoxia Zhang1, Jun Zheng1 and Honglin Jia1

1State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China

*These authors have contributed equally to this work

Correspondence to:

Honglin Jia, email: jiahonglin@caas.cn

Keywords: ClpB, caseinolytic proteases and chaperones, AAA+ ATPases, thermotolerance, toxoplasma gondii

Received: July 19, 2017    Accepted: August 27, 2017    Published: September 18, 2017

ABSTRACT

Caseinolytic peptidase B (ClpB) plays a pivotal role in suppressing and reversing protein aggregation. Toxoplasma gondii is an intracellular parasitic protozoan that infects a wide variety of mammals and birds and therefore is exposed to a broad range of living condition. We screened ToxoDB (http://ToxoDB.org) and identified 10 putative T. gondii genes encoding members of the Clp superfamily of caseinolytic proteases and chaperones. Of these, we focused on characterizing the Class I ATP-dependent molecular chaperones TgClpB1, TgClpB2, and TgClpB3. We found that TgClpB1, the most divergent of the five T. gondii Class I Clp ATPases, is cytoplasmic, TgClpB2 is found in the mitochondria of the parasites, and TgClpB3 is a ClpB with novel apicoplast localization. Knockout strains of TgClpB1 and TgClpB2 were established by CRISPR/Cas9 mutagenesis, and their complementing strains were constructed with FLAG-tag. Although knockout of TgClpB1 or TgClpB2 did not affect growth under normal circumstances, TgClpB1 was required for T. gondii thermotolerance. The growth, replication, and invasion capabilities of TgClpB1-deficient mutants were significantly inhibited after extracellular parasites were pretreated at 45°C. Moreover, TgClpB1 were observed at the poles of the ΔTgClpB1 FLAG-tagged strain treated at 42°C.


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