Transcription Factors BbPacC and Bbmsn2 Jointly Regulate Oosporein Production in Beauveria bassiana.
Transcription Factors BbPacC and Bbmsn2 Jointly Regulate Oosporein Production in Beauveria bassiana.
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DOI：
10.1128/spectrum.03118-22
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发表时间：
2022-12-21
影响因子：
3.7
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中科院分区：
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昆虫病原真菌球孢白僵菌可以在碱性条件下或真菌杀死的尸体中产生次生代谢物卵孢子素。然而，卵孢蛋白合成的调控机制尚不完全清楚。在本研究中，我们发现pH信号转录因子BbPacC参与了卵孢蛋白产生的调控。在pH 6.0或碱性（pH 8.0）条件下，过表达BbPacC可促进球孢白僵菌的卵孢素产生，但缺失该基因则会抑制卵孢素的产生。在酸性条件下（pH 4.0），野生型和BbPacC过表达菌株均未产生卵孢素。酵母单杂交实验和电泳迁移转移实验（EMSA）证实了BbPacC与4个假定的pacc结合位点在BbOpS3启动子区域的结合能力，BbOpS3是位于卵孢蛋白合成基因簇的转录因子，调节卵孢蛋白合成基因的表达。在所有测试条件下，在OEPacC或野生型菌株中，过表达Bbmsn2（先前报道的卵孢素合成负调节因子）可消除卵孢素的产生。然而，在BbPacC过表达菌株中，即使在pH为4.0时，删除Bbmsn2也显著提高了卵孢素的产量。这些结果表明，BbPacC是卵孢蛋白产生的正调节因子，并与Bbmsn2共同作用，在不同环境下，特别是在碱性条件下调节卵孢蛋白的产生。重要性球孢白僵菌在某些特定条件下产生红色二苯醌色素卵孢子素，如碱性条件和真菌杀死的尸体。卵孔蛋白具有抗菌和昆虫免疫抑制活性，在球孢白僵菌侵染昆虫宿主过程中发挥多种作用。一些与卵孢蛋白合成有关的负调节因子已被报道；然而，我们对生物合成基因簇之外的正调控因子知之甚少。在这里，我们发现pH信号转录因子BbPacC通过结合几个pacc结合位点正向调节卵孢蛋白的产生。此外，我们的研究结果还表明，BbPacC与负调节因子Bbmsn2共同调控卵孢蛋白的合成。我们的研究结果为了解球孢杆菌产卵孢子素的调控机制以及设计产高水平卵孢子素的球孢杆菌菌株提供了新的思路。
The entomopathogenic fungus Beauveria bassiana can produce the secondary metabolite oosporein under alkaline conditions or in fungus-killed cadavers. However, the regulatory mechanism of oosporein synthesis is not fully understood. In thisstudy, we found that the pH signaling transcription factor BbPacC is involved in the regulation of oosporein production. Overexpression of BbPacC promotes oosporein production in B. bassiana at pH 6.0 or under alkaline conditions (pH 8.0), but deletion of this gene abolished oosporein production. Under acidic conditions (pH 4.0), no oosporein production was observed in the wild-type and BbPacC overexpression strains. Yeast one-hybrid assays and electrophoretic mobility shift assay (EMSA) confirmed the binding ability of BbPacC with 4 putative PacC-binding sites in the promoter region of BbOpS3, a transcription factor located in the oosporein synthetic gene cluster regulating the expression of oosporein synthetic genes. Overexpression of Bbmsn2, a previously reported negative regulator of oosporein synthesis, in OEPacC or wild-type strains abolished oosporein production in all tested conditions. However, deletion of Bbmsn2 in the BbPacC overexpression strain significantly improved oosporein production even at pH 4.0. These results indicated that BbPacC is a positive regulator of oosporein production and functions jointly with Bbmsn2 to regulate oosporein production in different environments and particularly under alkaline conditions. IMPORTANCE B. bassiana produces the red dibenzoquinone pigment oosporein under certain specific conditions, such as alkaline conditions and fungus-killed cadavers. Ooporein possesses antibiotic and insect immune inhibition activities and plays multiple roles during the infection process of B. bassiana against insect hosts. Several negative regulators involved in oosporein synthesis have been reported; however, we know little about the positive regulators outside the biosynthetic gene cluster. Here, we found that the pH signaling transcription factor BbPacC positively regulates oosporein production by binding to several PacC-binding sites. In addition, our results also indicate that BbPacC jointly acts with the negative regulator Bbmsn2 to regulate oosporein synthesis. Our results provide insight into understanding the regulatory mechanism of oosporein production as well as targets to engineer B. bassiana strains producing high levels of oosporein.
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