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CRZ1/SP1样基因与致病性酵母新型隐球菌在有限通气条件下的生存、细胞完整性及生物膜形成相关。

The CRZ1/SP1-like gene links survival under limited aeration, cell integrity and biofilm formation in the pathogenic yeast Cryptococcus neoformans.

作者信息

Moranova Zuzana, Virtudazo Eric, Hricova Kristyna, Ohkusu Misako, Kawamoto Susumu, Husickova Vendula, Raclavsky Vladislav

机构信息

Department of Microbiology, Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic.

出版信息

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2014 Jun;158(2):212-20. doi: 10.5507/bp.2013.024. Epub 2013 Apr 22.

Abstract

AIMS

Limited aeration has been demonstrated to cause slowdown in proliferation and delayed budding, resulting eventually in a unique unbudded G2-arrest in the obligate aerobic pathogenic yeast Cryptococcus neoformans. Also, the ability to adapt to decreased oxygen levels during pathogenesis has been identified as a virulence factor in C. neoformans. The aim of this study was to identify and characterize genes that are necessary for the proliferation slowdown and G2-arrest caused by limited aeration.

METHODS

Random mutants were prepared and screened for lack of typical slowdown of proliferation under limited aeration. The CNAG_00156.2 gene coding for a zinc-finger transcription factor was identified in mutants showing most distinctive phenotype. Targeted deletion strain and reconstituted strain were prepared to characterize and confirm the gene functions. This gene was also identified in a parallel studies as homologous both to calcineurin responsive (Crz1) and PKC1-dependent (SP1-like) transcription factors.

RESULTS

We have confirmed the role of the cryptococcal homologue of CRZ1/SP1-like transcription factor in cell integrity, and newly demonstrated its role in slowdown of proliferation and survival under reduced aeration, in biofilm formation and in susceptibility to fluconazole.

CONCLUSIONS

Our data demonstrate a tight molecular link between slowdown of proliferation during hypoxic adaptation and maintenance of cell integrity in C. neoformans and present a new role for the CRZ1 family of transcription factors in fungi. The exact positioning of this protein in cryptococcal signalling cascades remains to be clarified.

摘要

目的

有限通气已被证明会导致增殖减缓及出芽延迟,最终致使专性需氧致病酵母新型隐球菌出现独特的无芽G2期阻滞。此外,在致病过程中适应降低的氧水平的能力已被确定为新型隐球菌的一种毒力因子。本研究的目的是鉴定和表征有限通气导致增殖减缓及G2期阻滞所必需的基因。

方法

制备随机突变体,并筛选在有限通气条件下缺乏典型增殖减缓的突变体。在表现出最显著表型的突变体中鉴定出编码锌指转录因子的CNAG_00156.2基因。制备靶向缺失菌株和重组菌株以表征和确认基因功能。在一项平行研究中,该基因也被鉴定为与钙调神经磷酸酶反应性(Crz1)和PKC1依赖性(SP1样)转录因子同源。

结果

我们已证实CRZ1/SP1样转录因子的隐球菌同源物在细胞完整性中的作用,并首次证明其在通气减少时的增殖减缓、存活、生物膜形成及对氟康唑敏感性方面的作用。

结论

我们的数据表明,新型隐球菌在低氧适应过程中的增殖减缓与细胞完整性维持之间存在紧密的分子联系,并揭示了转录因子CRZ1家族在真菌中的新作用。该蛋白在隐球菌信号级联反应中的准确定位仍有待阐明。

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