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Mas5, a homologue of bacterial DnaJ, is indispensable for the host infection and environmental adaptation of a filamentous fungal insect pathogen

机译:mas5, a homologue of bacterial DnaJ, is indispensable for the host infection and environmental adaptation of a filamentous fungal insect pathogen

摘要

Mas5, a yeast heat shock protein classified to the Hsp40 group, is homologous to bacterial archetype DnaJ but functionally unexplored in filamentous fungi. Here we identify a Mas5 homologue (46.86kDa) in Beauveria bassiana and show its indispensability for host infection and environmental adaptation of the fungal insect pathogen. The deletion of mas5 caused severe defects in aerial conidiation, conidial germination and submerged blastospore production (mimic to host haemocoel). The deletion mutant lost 100% virulence to Galleria mellonella larvae through normal cuticular penetration (topical inoculation) and 50% through cuticle-bypassing infection (intrahaemocoel injection). It formed no blastospore in vivo after inoculation or only a very few after injection. Its extracellular (cuticle degrading) enzymes and virulence-relating Pr1 proteases were 62% and 32% less active respectively. It became more sensitive to high osmolarity, oxidation, cell-wall perturbation, heat shock and UV-B irradiation. These concurred with reduced contents of intracellular mannitol and trehalose, decreased activities of antioxidant enzymes, impaired cell walls and suppressed transcripts of stress-responsive and virulence-relating genes. All the changes were restored by targeted mas5 complementation. All together, Mas5 is indispensable for the in vitro and in vivo life cycle of B.bassiana by targeting many sets of enzymes/proteins at transcriptional and post-transcriptional levels.
机译:Mas5是分类为Hsp40组的酵母热休克蛋白,与细菌原型DnaJ同源,但在丝状真菌中功​​能未开发。在这里,我们在球孢白僵菌中鉴定了Mas5同源物(46.86kDa),并显示了其对于宿主感染和真菌昆虫病原体的环境适应性必不可少。 mas5的缺失在空中分生孢子,分生孢子萌发和淹没的芽孢孢子生产(类似于宿主血红细胞)中造成了严重缺陷。缺失突变体通过正常的表皮穿透(局部接种)对梅花Gall幼虫失去了100%的毒力,而通过表皮旁路感染(输注血红素内)则失去了50%的毒力。接种后在体内或注射后仅形成极少量的芽孢。它的细胞外(表皮降解)酶和与毒力有关的Pr1蛋白酶的活性分别降低62%和32%。它对高渗透压,氧化,细胞壁微扰,热冲击和UV-B辐射变得更加敏感。这些与细胞内甘露醇和海藻糖含量降低,抗氧化酶活性降低,细胞壁受损以及应激反应性和毒力相关基因的转录物抑制有关。所有的变化都通过有针对性的mas5互补得以恢复。总之,通过在转录和转录后水平上靶向许多酶/蛋白质,Mas5对于黑僵菌的体外和体内生命周期都是必不可少的。

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