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Functional Analysis of atfA Gene to Stress Response in Pathogenic Thermal Dimorphic Fungus Penicillium marneffei

机译:atfA基因对病原性热双态真菌马尔尼菲青霉胁迫响应的功能分析

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摘要

Penicillium marneffei, the pathogenic thermal dimorphic fungus is a causative agent of a fatal systemic disease, penicilliosis marneffei, in immunocompromised patients especially HIV patients. For growth and survival, this fungus has to adapt to environmental stresses outside and inside host cells and this adaptation requires stress signaling pathways and regulation of gene expression under various kinds of stresses. In this report, P. marneffei activating transcription factor (atfA) gene encoding bZip-type transcription factor was characterized. To determine functions of this gene, atfA isogenic mutant strain was constructed using the modified split marker recombination method. The phenotypes and susceptibility to varieties of stresses including osmotic, oxidative, heat, UV, cell wall and cell membrane stresses of the mutant strain were compared with the wild type and the atfA complemented strains. Results demonstrated that the mRNA expression level of P. marneffei atfA gene increased under heat stress at 42°C. The atfA mutant was more sensitive to sodium dodecyl sulphate, amphotericin B and tert-butyl hydroperoxide than the wild type and complemented strains but not hydrogen peroxide, menadione, NaCl, sorbitol, calcofluor white, itraconazole, UV stresses and heat stress at 39°C. In addition, recovery of atfA mutant conidia after mouse and human macrophage infections was significantly decreased compared to those of wild type and complemented strains. These results indicated that the atfA gene was required by P. marneffei under specific stress conditions and might be necessary for fighting against host immune cells during the initiation of infection.
机译:马尔尼菲青霉菌是一种致病性热二形真菌,是免疫力低下的患者(尤其是HIV患者)致命性全身性疾病马尔尼菲青霉菌的致病因子。为了生长和存活,这种真菌必须适应宿主细胞内部和外部的环境压力,这种适应需要压力信号传导途径和在各种压力下的基因表达调控。在该报告中,表征了编码bZip型转录因子的马尼菲假单胞菌激活转录因子(atfA)基因。为了确定该基因的功能,使用改良的分裂标记重组方法构建了atfA等基因突变株。将突变菌株的表型和易感性与野生型和atfA互补菌株进行比较,包括渗透,氧化,热,紫外线,细胞壁和细胞膜应力。结果表明,在42°C的热胁迫下,马尔尼菲假单胞菌atfA基因的mRNA表达水平增加。 atfA突变体对十二烷基硫酸钠,两性霉素B和叔丁基氢过氧化物的敏感性高于野生型和互补菌株,但对过氧化氢,甲萘醌,NaCl,山梨糖醇,钙荧光白,伊曲康唑,UV胁迫和39°C的热胁迫则不敏感。 。此外,与野生型和互补菌株相比,小鼠和人类巨噬细胞感染后atfA突变体分生孢子的恢复显着降低。这些结果表明,在特定的应激条件下,马尔尼菲疟原虫需要atfA基因,并且可能是在感染开始时对抗宿主免疫细胞所必需的。

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