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雷氏蛋白调控真菌昆虫病原体在体外和体内无性循环所必需的营养代谢和运输。

Rei1-like protein regulates nutritional metabolism and transport required for the asexual cycle in vitro and in vivo of a fungal insect pathogen.

机构信息

MOE Laboratory of Biosystems Homeostasis & Protection, Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China.

College of Agricultural and Food Science, Zhejiang A&F University, Lin'an, Zhejiang 311300, China.

出版信息

Environ Microbiol. 2019 Aug;21(8):2772-2786. doi: 10.1111/1462-2920.14616. Epub 2019 Apr 22.

Abstract

Rei1 is a cytoplasm-specific pre-60S subunit export factor that functions exclusively in cold-sensitive yeast growth but remains unexplored in filamentous fungi. Here, we report that Rei1-like BbRei1 is localized in both cytoplasm and nucleus and acts as a vital regulator in Beauveria bassiana. Deletion of BbRei1 resulted in delayed conidial germination, abnormally polarized germlings, severe growth defects on various carbon/nitrogen sources and reduced conidiation capacity as well as low temperature-sensitive growth. In ΔBbrei1, greatly attenuated virulence correlated with reduced activities of enzymes secreted for cuticular penetration and blocked formation of hyphal bodies in vivo essential for facilitation of host mummification. Revealed by transcriptomic analysis, 560 and 840 genes were significantly up- and down-regulated in ΔBbrei1 versus wild-type respectively, representing 13.5% of the fungal genome. Many repressed genes were involved in metabolism and transport of carbohydrates and amino acids. However, electrophoretic mobility shift assays presented no interactions of purified BbRei1 with 14 promoter DNA fragments. Conclusively, BbRei1 plays a pivotal role in gene expression and metabolism of nutrients and energy essential for the asexual cycle in vitro and in vivo of B. bassiana and functions much beyond the role for the yeast Rei1 in cold-sensitive cell growth.

摘要

Rei1 是一种细胞质特异性的前 60S 亚基出口因子,仅在冷敏感酵母生长中发挥作用,但在丝状真菌中尚未被探索。在这里,我们报告 Rei1 样 BbRei1 定位于细胞质和细胞核中,并作为白僵菌的重要调节因子。BbRei1 的缺失导致分生孢子发芽延迟、异常极化的芽体、在各种碳/氮源上严重的生长缺陷以及产孢能力降低以及低温敏感生长。在ΔBbrei1 中,与降低的穿透表皮的酶活性相关的大大减弱的毒力以及体内丝状体的形成受阻,这对于促进宿主木乃伊化是必不可少的。通过转录组分析发现,与野生型相比,ΔBbrei1 中有 560 个和 840 个基因分别显著上调和下调,占真菌基因组的 13.5%。许多被抑制的基因参与碳水化合物和氨基酸的代谢和运输。然而,电泳迁移率变动分析显示纯化的 BbRei1 与 14 个启动子 DNA 片段之间没有相互作用。总之,BbRei1 在白僵菌无性周期的体外和体内的基因表达和营养物质代谢以及能量代谢中起着关键作用,其功能远远超出了酵母 Rei1 在冷敏感细胞生长中的作用。

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