The Cys2His2 zinc finger protein Zfp1 regulates sexual reproduction and virulence in Cryptococcus neoformans
The Cys2His2 zinc finger protein Zfp1 regulates sexual reproduction and virulence in Cryptococcus neoformans
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DOI：
10.1016/j.fgb.2019.01.002
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发表时间：
2019-03-01
影响因子：
3
通讯作者：
Liu, Tong-Bao
中科院分区：
文献类型：
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作者：
Fan, Cheng-Li;Han, Lian-Tao;Liu, Tong-Bao
新生隐球菌是一种普遍存在的酵母病原体，经常感染人类中枢神经系统(CNS)，导致免疫功能低下的人发生脑膜炎。虽然对真菌有性繁殖和毒力有重要作用的信号通路和因子已经被研究，但它们的确切作用机制仍有待进一步阐明。在这项研究中，我们鉴定并鉴定了一种新的锌指蛋白Zfp1，它调节新生隐孢子菌的有性繁殖和毒力。QRT-PCR和ZFP-1启动子调控活性分析表明，ZFP-1在交配过程中的各个时期都有表达。亚细胞定位分析表明，Zfp1定位于新生葡萄球菌的细胞质。体外胁迫反应分析表明，Zfp1Delta突变体和Zfp1高表达菌株Zfp1(OE)对SDS敏感，但对刚果红不敏感，表明Zfp1可能调节细胞膜的完整性。Zfp1对真菌的有性繁殖也是必不可少的，因为在zfp1 Delta突变体或Zfp1高表达菌株之间的双边交配中，担孢子的产生被阻止。真菌核发育分析表明，Zfp1高表达株系、突变株和Zfp1突变株在交配过程中的双侧核融合后不能进行减数分裂，表明Zfp1对交配过程中的减数分裂具有重要的调控作用。虽然zfp1 Delta突变体显示正常生长，并产生正常的主要毒力因子，但在小鼠模型中，毒力减弱。有趣的是，我们发现ZFP1过表达的菌株在小鼠全身感染模型中是无毒的。总体而言，我们的研究表明，锌指蛋白Zfp1对新生葡萄球菌的真菌产孢量和毒力是必不可少的。
Cryptococcus neoformans is a ubiquitous yeast pathogen that often infects the human central nervous system (CNS) to cause meningitis in immunocompromised individuals. Although numerous signaling pathways and factors important for fungal sexual reproduction and virulence have been investigated, their precise mechanism of action remains to be further elucidated. In this study, we identified and characterized a novel zinc finger protein Zfp1 that regulates fungal sexual reproduction and virulence in C. neoformans. qRT-PCR and ZFP1 promoter regulatory activity assays revealed a ubiquitous expression pattern of ZFP1 in all stages during mating. Subcellular localization analysis indicates that Zfp1 is targeted to the cytoplasm of C. neoformans. In vitro assays of stress responses showed that zfp1 Delta mutants and the ZFP1 overexpressed strains ZFP1(OE) are hypersensitive to SDS, but not Congo red, indicating that Zfp1 may regulate cell membrane integrity. Zfp1 is also essential for fungal sexual reproduction because basidiospore production was blocked in bilateral mating between zfp1 Delta mutants or ZFP1 overexpressed strains. Fungal nuclei development assay showed that nuclei in the bilateral mating of zfp1 Delta, mutants or ZFP1 overexpressed strains failed to undergo meiosis after fusion, indicating Zfp1 is important for regulating meiosis during mating. Although zfp1 Delta mutants showed normal growth and produced normal major virulence factors, virulence was attenuated in a murine model. Interestingly, we found that the ZFP1 overexpressed strains were avirulent in a murine systemic-infection model. Overall, our study showed that the zinc finger protein Zfp1 is essential for fungal sporulation and virulence in C. neoformans.