限制宿主死亡后细菌生长的卵孢霉素的调控级联反应和生物活性。

Regulatory cascade and biological activity of oosporein that limits bacterial growth after host death.

作者信息

Fan Yanhua, Liu Xi, Keyhani Nemat O, Tang Guirong, Pei Yan, Zhang Wenwen, Tong Sheng

机构信息

Biotechnology Research Center, Southwest University, Chongqing 400715, China;

Biotechnology Research Center, Southwest University, Chongqing 400715, China.

出版信息

Proc Natl Acad Sci U S A. 2017 Feb 28;114(9):E1578-E1586. doi: 10.1073/pnas.1616543114. Epub 2017 Feb 13.

Abstract

The regulatory network and biological functions of the fungal secondary metabolite oosporein have remained obscure. has evolved the ability to parasitize insects and outcompete microbial challengers for assimilation of host nutrients. A novel zinc finger transcription factor, BbSmr1 ( secondary metabolite regulator 1), was identified in a screen for oosporein overproduction. Deletion of resulted in up-regulation of the oosporein biosynthetic gene cluster ( genes) and constitutive oosporein production. Oosporein production was abolished in double mutants of and a second transcription factor, , within the oosporein gene cluster (), indicating that BbSmr1 acts as a negative regulator of expression. Real-time quantitative PCR and a GFP promoter fusion construct of , the oosporein polyketide synthase, indicated that is expressed mainly in insect cadavers at 24-48 h after death. Bacterial colony analysis in -infected insect hosts revealed increasing counts until host death, with a dramatic decrease (∼90%) after death that correlated with oosporein production. In vitro studies verified the inhibitory activity of oosporein against bacteria derived from insect cadavers. These results suggest that oosporein acts as an antimicrobial compound to limit microbial competition on -killed hosts, allowing the fungus to maximally use host nutrients to grow and sporulate on infected cadavers.

摘要

真菌次级代谢产物卵孢素的调控网络和生物学功能一直不清楚。[真菌名称未给出]已经进化出寄生昆虫并在宿主养分同化方面胜过微生物竞争者的能力。在一次卵孢素过量生产筛选中鉴定出一种新型锌指转录因子BbSmr1(次级代谢产物调节因子1)。BbSmr1的缺失导致卵孢素生物合成基因簇(基因)上调以及卵孢素的组成型产生。在卵孢素基因簇()内BbSmr1和第二个转录因子[第二个转录因子名称未给出]的双突变体中卵孢素产生被消除,这表明BbSmr1作为卵孢素基因簇表达的负调节因子。卵孢素聚酮合酶基因的实时定量PCR和绿色荧光蛋白启动子融合构建体表明该基因主要在昆虫尸体死亡后24 - 48小时表达。在被[真菌名称未给出]感染的昆虫宿主中进行的细菌菌落分析显示,直到宿主死亡细菌数量增加,死亡后细菌数量急剧下降(约90%),这与卵孢素产生相关。体外研究证实了卵孢素对源自昆虫尸体的细菌的抑制活性。这些结果表明,卵孢素作为一种抗菌化合物限制了在[真菌名称未给出]杀死的宿主上的微生物竞争,使真菌能够最大限度地利用宿主养分在受感染的尸体上生长和产孢。

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