NcGRA17 is an important regulator of parasitophorous vacuole morphology and pathogenicity of Neospora caninum

文献类型： 外文期刊

第一作者： Yang, Congshan

作者： Yang, Congshan;Liu, Jing;Ma, Lei;Zhang, Xiao;Zhou, Bingxin;Liu, Qun;Yang, Congshan;Liu, Jing;Ma, Lei;Zhang, Xiao;Zhou, Bingxin;Liu, Qun;Zhang, Xichen;Zhu, Xingquan

作者机构：

关键词： Neospora caninum; CRISPR/cas9; Dense granule protein 17; Parasitophorous vacuole; Pathogenicity

期刊名称：VETERINARY PARASITOLOGY （ 影响因子：2.738； 五年影响因子：2.951 ）

ISSN： 0304-4017

年卷期： 2018 年 264 卷

页码：

收录情况： SCI

摘要： Neospora caninum is an obligate intracellular protozoan parasite that infects a wide range of mammalian species, and particularly causes the reproductive loss in cattle. We identified a novel dense granule protein, N. caninum granule protein 17 (NcGRA17) using the CRISPR/cas9 genome editing system and studied its function. We generated the NcGRA17 knockout strain (Delta NcGRA17) and NcGRA17 complementary strain (i Delta NcGRA17). Plaque assays and intracellular proliferation tests showed that the Delta NcGRA17 strain formed smaller plaques and had slower intracellular growth. Mouse virulence assay showed loss of virulence for the Delta NcGRA17 strain. We observed that the parasitophorous vacuoles (PVs) of NcGRA17-deficient parasites have aberrant morphology. To investigate the contribution of NcGRA17 alpha-helices to aberrant morphology of PVs, we transfected four truncated forms of NcGRA17 into NcGRA17 knockout strain and the phenotypes of these mutants were analysed. Lack of the N-terminal region (NT) failed to target the protein to dense granules, while NcGRA17 (Delta alpha 1)-HA, NcGRA17 (Delta alpha 2-4)-HA and NcGRA17 (Delta alpha 5-8)-HA were targeted to dense granules, but failed to rescue the aberrant PV morphology. Our results indicate that NcGRA17 as a dense granule protein determines PV morphology and pathogenicity, and alpha-helices of NcGRA17 may be responsible for the aberrant morphology of N. caninum PVs.

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