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核输入蛋白 MoKap119 介导的细胞周期蛋白依赖性激酶调控因子 MoCks1 的核输入对于稻瘟病菌的致病性至关重要。

Karyopherin MoKap119-mediated nuclear import of cyclin-dependent kinase regulator MoCks1 is essential for Magnaporthe oryzae pathogenicity.

机构信息

State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangzhou, China.

Integrative Microbiology Research Centre/Guangdong Province Key Laboratory of Microbial Signals and Disease Control, South China Agricultural University, Guangzhou, China.

出版信息

Cell Microbiol. 2020 Jan;22(1):e13114. doi: 10.1111/cmi.13114. Epub 2019 Oct 9.

Abstract

Nuclear import of proteins relies on nuclear import receptors called importins/karyopherins (Kaps), whose functions were reported in yeasts, fungi, plants, and animal cells, including cell cycle control, morphogenesis, stress sensing/response, and also fungal pathogenecity. However, limited is known about the physiological function and regulatory mechanism of protein import in the rice-blast fungus Magnaporthe oryzae. Here, we identified an ortholog of β-importin in M. oryzae encoded by an ortholog of KAP119 gene. Functional characterisation of this gene via reverse genetics revealed that it is required for vegetative growth, conidiation, melanin pigmentation, and pathogenicity of M. oryzae. The mokap119Δ mutant was also defective in formation of appressorium-like structure from hyphal tips. By affinity assay and liquid chromatography-tandem mass spectrometry, we identified potential MoKap119-interacting proteins and further verified that MoKap119 interacts with the cyclin-dependent kinase subunit MoCks1 and mediates its nuclear import. Transcriptional profiling indicated that MoKap119 may regulate transcription of infection-related genes via MoCks1 regulation of MoSom1. Overall, our findings provide a novel insight into the regulatory mechanism of M. oryzae pathogenesis likely by MoKap119-mediated nuclear import of the cyclin-dependent kinase subunit MoCks1.

摘要

蛋白质的核输入依赖于称为核输入受体的核输入受体,也称为输入蛋白/核孔蛋白(Kaps),其功能在酵母、真菌、植物和动物细胞中已有报道,包括细胞周期调控、形态发生、应激感应/反应,以及真菌的致病性。然而,关于水稻稻瘟病菌(Magnaporthe oryzae)中蛋白质输入的生理功能和调节机制知之甚少。在这里,我们鉴定了 M. oryzae 中β-输入蛋白的同源物,由 KAP119 基因的同源物编码。通过反向遗传学对该基因的功能表征表明,它是 M. oryzae 营养生长、分生孢子形成、黑色素沉着和致病性所必需的。mokap119Δ突变体也在菌丝尖端形成类似附着胞的结构方面存在缺陷。通过亲和测定和液相色谱-串联质谱分析,我们鉴定了潜在的 MoKap119 相互作用蛋白,并进一步验证了 MoKap119 与细胞周期蛋白依赖性激酶亚基 MoCks1 相互作用,并介导其核输入。转录谱分析表明,MoKap119 可能通过 MoCks1 调节 MoSom1 来调节感染相关基因的转录。总之,我们的研究结果为 M. oryzae 发病机制的调节机制提供了新的见解,可能是通过 MoKap119 介导的细胞周期蛋白依赖性激酶亚基 MoCks1 的核输入。

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