title: Identification and characterization of the verticillin biosynthetic gene cluster in Clonostachys rogersoniana.
abstract: Verticillin is one of the dimeric epipolythiodioxopiperazines (ETPs) which are toxic secondary metabolites produced only by fungi. ETPs have received substantial attention since its complex molecular architecture and a wide range of biological activities. Although biosynthesis of the monomeric gliotoxin has been studied extensively, the biosynthetic pathway of dimeric ETPs is far from being studied. To investigate the biosynthesis of dimeric ETPs and expand our understanding of their dimerization, the verticillin biosynthetic gene cluster (ver) was identified and cloned from a genomic DNA fosmid library of the Cordyceps-colonizing fungus Clonostachys rogersoniana with the designed primers based on the sequence of a nonribosomal peptide synthetase (NRPS) ChaP which was predicted to be responsible for chaetocin biosynthesis in Chaetomium virescens. To validate it, the chaP homologous gene verP in the ver cluster was disrupted. HPLC-MS analysis demonstrated that the verP disruption mutant (ΔverP) completely abolished verticillin production, and it could be restored by introducing a copy of the wild-type verP gene. Further gene disruptions and chemical analysis demonstrated that most genes of this ver cluster were essential for verticillin biosynthesis. Intriguingly, disruption of verP almost abolished the conidiation of Clonostachys rogersoniana and it was partially restored by addition of the fermentation extract which contains verticillin, implying that verticillin or its intermediate plays a role in the Cordyceps-colonizing fungal morphological differentiation.
authors.affiliation: State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Pengjie
authors.initials: P
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Pan
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Yaxin
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Xingzhong
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Toxicology & Medical Countermeasures, Beijing Institute of Pharmacology & Toxicology, Beijing 100850, China.
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Che
authors.affiliation: State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address: liug@im.ac.cn.
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics; D006999:Hypocreales / Q000235:genetics* / Q000378:metabolism; D007211:Indoles / Q000378:metabolism / Q000633:toxicity; D005810:Multigene Family / Q000235:genetics; D010453:Peptide Synthases / Q000235:genetics*; D010879:Piperazines / Q000378:metabolism
chemical_list: D007211:Indoles; D010879:Piperazines; C009654:verticillins; C002511:chaetocin; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase
keywords: Clonostachys rogersoniana; Conidiation; Verticillin biosynthesis cluster; verP