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Sho1 在烟曲霉极性生长中的作用。

The role of sho1 in polarized growth of Aspergillus fumigatus.

机构信息

Peking University First Hospital, Research Center for Medical Mycology, Peking University, No. 8, Xishiku Street, West District, Beijing, 100034, People's Republic of China.

出版信息

Mycopathologia. 2011 Nov;172(5):347-55. doi: 10.1007/s11046-011-9452-4. Epub 2011 Jul 28.

Abstract

Aspergillus fumigatus is an opportunistic pathogen that may cause severe invasive disease in immunocompromised patients. The filamentous fungi undergo polarized growth, searching for nutrients in the environment and causing invasive growth in tissue. Sho1 is a sensor of the high osmolarity glycerol pathway, and the sho1 mutant showed a decrease in growth rate. We found that sho1 is involved in the polarized growth of A. fumigatus. The sho1 mutation resulted in extended isotropic growth of germinating conidia followed by multiple germ tubes and wide hyphae with short intercalary cells by calcofluor white staining. The mechanism by which sho1 gene affected polarized growth is investigated. A reduced number of apical vesicles with greater dispersion were observed by transmission electron microscopy in the Spitzenkörper body of the sho1 mutant. Actin patches were distributed randomly at low density at early stages of mutant strain fungal development and reaggregated to the hyphal tip of later stages when long filamentous fungi formed. Actin patches located at the tip of polarized wild-type cells. RNA levels of polarized growth-related genes Rho GTPases were detected by real-time PCR. The sho1 gene did not affect the RNA expression when strains were cultured at 37°C for 6 h. At 17 h, the RNA expression of rho1, rho3 and CDC42 in the sho1 mutant were 0.18-, 0.18- and 0.33-fold of that in the wild type. The sho1 gene affected the polarized growth through affecting the expression of Rho GTPases, the distribution of actin cytoskeleton, vesicle quantity and distribution.

摘要

烟曲霉是一种机会性病原体,可导致免疫功能低下的患者发生严重的侵袭性疾病。丝状真菌经历极性生长,在环境中寻找营养物质,并在组织中引起侵袭性生长。Sho1 是高渗透压甘油途径的传感器,sho1 突变体的生长速度下降。我们发现 sho1 参与了烟曲霉的极性生长。sho1 突变导致萌发的分生孢子的各向同性生长延长,随后是多个发芽管和宽的菌丝,短的间插细胞通过钙荧光白染色。研究了 sho1 基因影响极性生长的机制。透射电子显微镜观察到 sho1 突变体的 Spitzenkörper 体中顶体小泡数量减少,分散度增加。在突变体菌株真菌发育的早期阶段,肌动蛋白斑以低密度随机分布,当形成长丝状真菌时,它们重新聚集到菌丝尖端。肌动蛋白斑位于极性野生型细胞的尖端。通过实时 PCR 检测与极性生长相关的 Rho GTPases 基因的 RNA 水平。当菌株在 37°C 下培养 6 小时时,sho1 基因不影响 RNA 表达。在 17 小时时,sho1 突变体中 rho1、rho3 和 CDC42 的 RNA 表达分别为野生型的 0.18-、0.18-和 0.33 倍。sho1 基因通过影响 Rho GTPases 的表达、肌动蛋白细胞骨架的分布、囊泡数量和分布来影响极性生长。

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