Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, 310058, China.
Shandong Provincial Key Laboratory of Microbial Engineering, Qilu University of Technology, Jinan, Shandong, 250353, China.
Sci Rep. 2017 Mar 15;7:44360. doi: 10.1038/srep44360.
Lysine acetylation (Kac) events in filamentous fungi are poorly explored. Here we show a lysine acetylome generated by LC-MS/MS analysis of immunoaffinity-based Kac peptides from normal hyphal cells of Beauveria bassiana, a fungal entomopathogen. The acetylome comprised 283 Kac proteins and 464 Kac sites. These proteins were enriched to eight molecular functions, 20 cellular components, 27 biological processes, 20 KEGG pathways and 12 subcellular localizations. All Kac sites were characterized as six Kac motifs, including a novel motif (KacW) for 26 Kac sites of 17 unknown proteins. Many Kac sites were predicted to be multifunctional, largely expanding the fungal Kac events. Biological importance of identified Kac sites was confirmed through functional analysis of Kac sites on Pmt1 and Pmt4, two O-mannosyltransferases. Singular site mutations (K88R and K482R) of Pmt1 resulted in impaired conidiation, attenuated virulence and decreased tolerance to oxidation and cell wall perturbation. These defects were close to or more severe than those caused by the deletion of pmt1. The Pmt4 K360R mutation facilitated colony growth under normal and stressful conditions and enhanced the fungal virulence. Our findings provide the first insight into the Kac events of B. bassiana and their links to the fungal potential against insect pests.
丝状真菌中的赖氨酸乙酰化(Kac)事件研究甚少。在这里,我们通过 LC-MS/MS 分析免疫亲和法提取的白僵菌正常菌丝细胞中的 Kac 肽,展示了一个赖氨酸乙酰基组。乙酰基组包括 283 个 Kac 蛋白和 464 个 Kac 位点。这些蛋白被富集到八个分子功能、20 个细胞成分、27 个生物过程、20 个 KEGG 途径和 12 个亚细胞定位。所有的 Kac 位点都被表征为六个 Kac 基序,包括一个新的基序(KacW),对应 17 个未知蛋白的 26 个 Kac 位点。许多 Kac 位点被预测为多功能性的,这极大地扩展了真菌的 Kac 事件。通过对 Pmt1 和 Pmt4 上 Kac 位点的功能分析,证实了鉴定出的 Kac 位点的生物学重要性。Pmt1 的单个位点突变(K88R 和 K482R)导致分生孢子形成受损、毒力减弱以及对氧化和细胞壁扰动的耐受性降低。这些缺陷与 pmt1 缺失引起的缺陷相似或更严重。Pmt4 的 K360R 突变促进了正常和应激条件下的菌落生长,并增强了真菌的毒力。我们的研究结果首次揭示了白僵菌的 Kac 事件及其与真菌对昆虫害虫的潜在作用之间的联系。