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首页> 外文期刊>Veterinary Parasitology >NcGRA17 is an important regulator of parasitophorous vacuole morphology and pathogenicity of Neospora caninum
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NcGRA17 is an important regulator of parasitophorous vacuole morphology and pathogenicity of Neospora caninum

机译：NcGra17是Neospora caninum的寄生液态和致病性的重要调节因子

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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.
机译：Neospora caninum是一种迫使细胞内原生动物寄生虫，其感染了各种哺乳动物物种，特别是牛的生殖损失。我们鉴定了一种使用CRISPR / CAS9基因组编辑系统的新型致密颗粒蛋白，N，Caninum颗粒蛋白17（NcGra17）并研究其功能。我们产生了Ncgra17敲除菌株（Delta Ncgra17）和Ncgra17互补菌株（I delta ncgra17）。斑块测定和细胞内增殖试验表明，Delta Ncgra17菌株形成较小的斑块并具有较慢的细胞内生长。小鼠毒力测定显示Delta Ncgra17菌株的毒力丧失。我们观察到NCGRA17缺陷寄生虫的寄生泡（PVS）具有异常形态。为了探讨NCGRA17α-螺旋对PV的异常形态的贡献，我们将四种截短的形式的NCGRA17转染到NCGRA17敲除菌株中，并分析这些突变体的表型。缺乏N-末端区域（NT）未能将蛋白质靶向致密颗粒，而Ncgra17（Delta alpha1）-Ha，Ncgra17（Delta alpha 2-4）-Ha和Ncgra17（Delta alpha 5-8）-ha靶向致密颗粒，但未能拯救异常的PV形态。我们的结果表明，NCGRA17作为致密颗粒蛋白质确定PV形态和致病性，NCGRA17的α-螺旋可以是N.Caninum PVS的异常形态。

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