Lysine 2-hydroxyisobutyrylation(Khib) is a newly identified post-translational modification(PTM) that plays important roles in transcription and cell proliferation in eukaryotes. However, its function remains unknown ...Lysine 2-hydroxyisobutyrylation(Khib) is a newly identified post-translational modification(PTM) that plays important roles in transcription and cell proliferation in eukaryotes. However, its function remains unknown in phytopathogenic fungi. Here,we performed a comprehensive assessment of Khibin the rice false smut fungus Ustilaginoidea virens, using Tandem Mass Tag(TMT)-based quantitative proteomics approach. A total of 3 426 Khibsites were identified in 977 proteins, sugg esting that Khibis a common and complex PTM in U. virens. Our data demonstrated that the2-hydroxyisobutyrylated proteins are involved in diverse biological processes. Network analysis of the modified proteins revealed a highly interconnected protein network that included many well-studied virulence factors. We confirmed that the Zn-binding reduced potassium dependency3-type histone deacetylase(UvRpd3) is a major enzyme that removes 2-hydroxyisobutyrylation and acetylation in U. virens. Notably, mutations of Khib sites in the mitogen-activated protein kinase(MAPK)UvSlt2 significantly reduced fungal virulence and decreased the enzymatic activity of UvSlt2. Molecular dynamics simulations demonstrated that 2-hydroxyisobutyrylation in UvSlt2 increased the hydrophobic solvent-accessible surface area and thereby affected binding between the UvSlt2 enzyme and its substrates. Our findings thus establish Khibas a major post-translational modification in U. virens and point to an important role for Khibin the virulence of this phytopathogenic fungus.展开更多
Microbes serve as the most important resource for drug discovery.During our screening for bioactive compounds from our natural products library,a pathogenic fungus,Microdochium majus strain 99049,from wheat was select...Microbes serve as the most important resource for drug discovery.During our screening for bioactive compounds from our natural products library,a pathogenic fungus,Microdochium majus strain 99049,from wheat was selected for further investigation.A new alkaloid named brocaeloid D(1),together with six previously characterized compounds(2–7)were identified.Compound 1 belongs to 4-oxoquinoline with C-2 reversed prenylation and a succinimide substructure.All the structures of these newly isolated compounds were determined by different means in spectroscopic experiments.The absolute configurations of 1 was further deduced from comparison of its CD spectrum with that of known compound 2.The bioactivities of these identified compounds were evaluated against several pathogenic microorganisms and cancer cell lines.Compounds 1–5 showed activity against HUH-7 human hepatoma cells with IC50 values of 80μg/mL.Compound 6 showed mild activity against HeLa cells(IC50=51.9μg/mL),weak anti-MTB activity(MIC=80μg/mL),and moderate anti-MRSA activity(MIC=25μg/mL),and compound 7 showed weak anti-MRSA activity(MIC=100μg/mL).展开更多
The chemical diversity of terpenoids is typically established by terpene synthase-catalyzed cyclization and diversified by post-tailoring modifications.Fungal bifunctional terpene synthase(BFTS)associated P450 enzymes...The chemical diversity of terpenoids is typically established by terpene synthase-catalyzed cyclization and diversified by post-tailoring modifications.Fungal bifunctional terpene synthase(BFTS)associated P450 enzymes have shown significant catalytic potentials through the development of various new terpenoids with different biological activities.This study discovered the BFTS and its related gene cluster from the plant endophytic fungus Didymosphaeria variabile 17020.Heterologous expression of the BFTS in Saccharomyces cerevisiae resulted in the characterization of a major product diterpene variediene(1),along with two new minor products neovariediene and neoflexibilene.Further heterologous expression of the BFTS and one cytochrome P450 enzyme VndE(CYP6138B1)in Aspergillus oryzae NSAR1 led to the identification of seven norditerpenoids(19 carbons)with a structurally unique 5/5 bicyclic ring system.Interestingly,in vivo experiments suggested that the cyclized terpene variediene(1)was modified by VndE along with the endogenous enzymes from the host cell A.oryzae through serial chemical conversions,followed by multi-site hydroxylation via A.oryzae endogenous enzymes.Our work revealed that the two-enzymes biosynthetic system and host cell machinery could produce structurally unique terpenoids.展开更多
Phytopathogenic fungi have attracted great attention as a promising source for new drug discovery.In the progress of our ongoing study for bioactive natural products from an in-house phytopathogenic fungi library,a pa...Phytopathogenic fungi have attracted great attention as a promising source for new drug discovery.In the progress of our ongoing study for bioactive natural products from an in-house phytopathogenic fungi library,a pathogenic fungus,Fusarium proliferatum strain 13294(FP13294),was selected for chemical investigation.Two novel aliphatic unsaturated alcohols named fusariumnols A and B(1 and 2),together with one previously characterized sesquiterpenoid lignoren(3)were identified.Structures of 1-3 were assigned by mass spectrometry and NMR spectroscopy.Their bioactivities were assessed against Staphylococcus epidermidis,S.aureus,and Methicillin-resistant S.aureus(MRSA).Compounds 1 and 2 exhibited weak antibacterial activity against S.epidermidis(MIC=100μM).展开更多
基金supported by the National Natural Science Foundation of China (32072371)the National Key Research and Development Program (2016YFD0300700, 2017YFD0301400)the Fundamental Research Funds for the Central Universities of China (2662018JC051)。
文摘Lysine 2-hydroxyisobutyrylation(Khib) is a newly identified post-translational modification(PTM) that plays important roles in transcription and cell proliferation in eukaryotes. However, its function remains unknown in phytopathogenic fungi. Here,we performed a comprehensive assessment of Khibin the rice false smut fungus Ustilaginoidea virens, using Tandem Mass Tag(TMT)-based quantitative proteomics approach. A total of 3 426 Khibsites were identified in 977 proteins, sugg esting that Khibis a common and complex PTM in U. virens. Our data demonstrated that the2-hydroxyisobutyrylated proteins are involved in diverse biological processes. Network analysis of the modified proteins revealed a highly interconnected protein network that included many well-studied virulence factors. We confirmed that the Zn-binding reduced potassium dependency3-type histone deacetylase(UvRpd3) is a major enzyme that removes 2-hydroxyisobutyrylation and acetylation in U. virens. Notably, mutations of Khib sites in the mitogen-activated protein kinase(MAPK)UvSlt2 significantly reduced fungal virulence and decreased the enzymatic activity of UvSlt2. Molecular dynamics simulations demonstrated that 2-hydroxyisobutyrylation in UvSlt2 increased the hydrophobic solvent-accessible surface area and thereby affected binding between the UvSlt2 enzyme and its substrates. Our findings thus establish Khibas a major post-translational modification in U. virens and point to an important role for Khibin the virulence of this phytopathogenic fungus.
基金This work was partially supported by the grants from the National Natural Science Foundation of China(31430002,81573341,21877038,31720103901,31320103911)Taishan Scholarship,Open Project Funding of the State Key Laboratory of Bioreactor Engineering,the 111 Project(B18022)+1 种基金National Key R&D Program of China 2017YFE0108200the Fundamental Research Funds for the Central Universities(22221818014).
文摘Microbes serve as the most important resource for drug discovery.During our screening for bioactive compounds from our natural products library,a pathogenic fungus,Microdochium majus strain 99049,from wheat was selected for further investigation.A new alkaloid named brocaeloid D(1),together with six previously characterized compounds(2–7)were identified.Compound 1 belongs to 4-oxoquinoline with C-2 reversed prenylation and a succinimide substructure.All the structures of these newly isolated compounds were determined by different means in spectroscopic experiments.The absolute configurations of 1 was further deduced from comparison of its CD spectrum with that of known compound 2.The bioactivities of these identified compounds were evaluated against several pathogenic microorganisms and cancer cell lines.Compounds 1–5 showed activity against HUH-7 human hepatoma cells with IC50 values of 80μg/mL.Compound 6 showed mild activity against HeLa cells(IC50=51.9μg/mL),weak anti-MTB activity(MIC=80μg/mL),and moderate anti-MRSA activity(MIC=25μg/mL),and compound 7 showed weak anti-MRSA activity(MIC=100μg/mL).
基金the financial support from the National Key Research and Development Program of China(2020YFA0907800 and 2019YFA0906200)the National Natural Science Foundation of China(21907031,81903529,21977029,31720103901,21877124)+2 种基金the Open Project Funding of the State Key Laboratory of Bioreactor Engineeringthe 111 Project(B18022)Genome sequencing and assembly of strain DV17020 were supported by funding from the Natural Science and Engineering Research Council of Canada to Prof.T.Hsiang.
文摘The chemical diversity of terpenoids is typically established by terpene synthase-catalyzed cyclization and diversified by post-tailoring modifications.Fungal bifunctional terpene synthase(BFTS)associated P450 enzymes have shown significant catalytic potentials through the development of various new terpenoids with different biological activities.This study discovered the BFTS and its related gene cluster from the plant endophytic fungus Didymosphaeria variabile 17020.Heterologous expression of the BFTS in Saccharomyces cerevisiae resulted in the characterization of a major product diterpene variediene(1),along with two new minor products neovariediene and neoflexibilene.Further heterologous expression of the BFTS and one cytochrome P450 enzyme VndE(CYP6138B1)in Aspergillus oryzae NSAR1 led to the identification of seven norditerpenoids(19 carbons)with a structurally unique 5/5 bicyclic ring system.Interestingly,in vivo experiments suggested that the cyclized terpene variediene(1)was modified by VndE along with the endogenous enzymes from the host cell A.oryzae through serial chemical conversions,followed by multi-site hydroxylation via A.oryzae endogenous enzymes.Our work revealed that the two-enzymes biosynthetic system and host cell machinery could produce structurally unique terpenoids.
基金This work was supported by the National Key Research and Development Program of China(2020YFA0907200,2019YFA0906200,and 2020YFA0907800)the National Natural Science Foundation of China(21877038,21907031,21977029,31720103901,and 81903529)+1 种基金Shanghai Rising-Star Program(20QA1402800)the Open Project Funding of the State Key Laboratory of Bioreactor Engineering,and the 111 Project(B18022).
文摘Phytopathogenic fungi have attracted great attention as a promising source for new drug discovery.In the progress of our ongoing study for bioactive natural products from an in-house phytopathogenic fungi library,a pathogenic fungus,Fusarium proliferatum strain 13294(FP13294),was selected for chemical investigation.Two novel aliphatic unsaturated alcohols named fusariumnols A and B(1 and 2),together with one previously characterized sesquiterpenoid lignoren(3)were identified.Structures of 1-3 were assigned by mass spectrometry and NMR spectroscopy.Their bioactivities were assessed against Staphylococcus epidermidis,S.aureus,and Methicillin-resistant S.aureus(MRSA).Compounds 1 and 2 exhibited weak antibacterial activity against S.epidermidis(MIC=100μM).
