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蛋白磷酸酶基因 MaPpt1 作为金龟子绿僵菌微轮回产孢的编程因子和 UV-B 耐受性的负调控因子发挥作用。

The protein phosphatase gene MaPpt1 acts as a programmer of microcycle conidiation and a negative regulator of UV-B tolerance in Metarhizium acridum.

机构信息

Genetic Engineering Research Center, School of Life Sciences, Chongqing University, Chongqing, 401331, People's Republic of China.

Chongqing Engineering Research Center for Fungal Insecticide, Chongqing, 401331, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2019 Feb;103(3):1351-1362. doi: 10.1007/s00253-018-9567-3. Epub 2019 Jan 4.

DOI:10.1007/s00253-018-9567-3
PMID:30610282
Abstract

The Ser/Thr protein phosphatase Ppt1 (yeast)/PP5 (humans) has been implicated in signal transduction-mediated growth and differentiation, DNA damage/repair, cell cycle progression, and heat shock responses. Little, however, is known concerning the functions of Ppt1/PP5 in filamentous fungi. In this study, the Ppt1 gene MaPpt1 was characterized in the insect pathogenic fungus, Metarhizium acridum. The MaPpt1 protein features a three-tandem tetratricopeptide repeat (TPR) domain and a peptidyl-prolyl cis-trans isomerase-like (PP2Ac) domain. Subcellular localization using an MaPpt1::eGFP fusion protein revealed that MaPpt1 was localized in the cytoplasm of spores, but gathered at the septa in growing hyphae. Targeted gene inactivation of MaPpt1 in M. acridum resulted in unexpected reprogramming of normal aerial conidiation to microcycle conidiation. Although overall vegetative growth was unaffected, a significant increase in conidial yield was noted in ΔMaPpt1. Stress-responsive phenotypes and virulence were largely unaffected in ΔMaPpt1. Exceptionally, ΔMaPpt1 displayed increased UV tolerance compared to wild type. Digital gene expression data revealed that MaPpt1 mediates transcription of sets of genes involved in conidiation, polarized growth, cell cycle, cell proliferation, DNA replication and repair, and some important signaling pathways. These data indicate a unique role for Ppt1 in filamentous fungal development and differentiation.

摘要

丝氨酸/苏氨酸蛋白磷酸酶 Ppt1(酵母)/PP5(人类)已被牵连到信号转导介导的生长和分化、DNA 损伤/修复、细胞周期进程和热休克反应中。然而,关于 Ppt1/PP5 在丝状真菌中的功能知之甚少。在这项研究中,昆虫病原真菌绿僵菌中的 Ppt1 基因 MaPpt1 被进行了特征描述。MaPpt1 蛋白具有三个串联四肽重复(TPR)结构域和一个肽基脯氨酰顺反异构酶样(PP2Ac)结构域。使用 MaPpt1::eGFP 融合蛋白进行亚细胞定位表明,MaPpt1 定位于孢子的细胞质中,但在生长菌丝的隔膜处聚集。在绿僵菌中靶向基因灭活 MaPpt1 导致正常气生分生孢子异常重编程为微周期分生孢子。尽管总体营养生长不受影响,但ΔMaPpt1 中的分生孢子产量显著增加。ΔMaPpt1 中的应激反应表型和毒力基本不受影响。例外的是,与野生型相比,ΔMaPpt1 显示出更高的 UV 耐受性。数字基因表达数据显示,MaPpt1 介导与分生孢子形成、极化生长、细胞周期、细胞增殖、DNA 复制和修复以及一些重要信号通路相关的一组基因的转录。这些数据表明 Ppt1 在丝状真菌发育和分化中具有独特的作用。

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