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
The post-translational modification lysine 2-hydroxyisobutyrylation(K_(hib))plays an important role in gene transcription,metabolism,and enzymatic activity.K_(hib)sites have been identified in rice(Oryza sativa).Howev...The post-translational modification lysine 2-hydroxyisobutyrylation(K_(hib))plays an important role in gene transcription,metabolism,and enzymatic activity.K_(hib)sites have been identified in rice(Oryza sativa).However,the K_(hib)status of proteins in rice flowers during pathogen infection remains unclear.Here,we report a comprehensive identification of K_(hib)-modified proteins in rice flowers,and the changes in these proteins during infection with the fungal pathogen Ustilaginoidea virens.By using a tandem mass tag-based quantitative proteomics approach,we identified 2,891 K_(hib)sites on 964 proteins in rice flowers.Our data demonstrated that 2-hydroxyisobutyrylated proteins are involved in diverse biological processes.K_(hib)levels were substantially reduced upon infection with U.virens.Chromatin immunoprecipitation polymerase chain reaction(PCR)and reverse transcription quantitative PCR analyses revealed that histone K_(hib)is involved in the expression of disease-resistance genes.More importantly,most quantified sites on core histones H3 were downregulated upon U.virens infection.In addition,the histone deacetylases HDA705,HDA716,SRT1,and SRT2 are involved in the removal of K_(hib)marks in rice.HDA705 was further confirmed to negatively regulate rice disease resistance to pathogens U.virens,Magnaporthe oryzae,and Xanthomonas oryzae pv.oryzae(Xoo).Our data suggest that U.virens could modulate K_(hib)in rice flowers during infection.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 supported by the National Natural Science Foundation of China(32072371)the National Key Research and Development Program(2016YFD0300700)the Fundamental Research Funds for the Central Universities of China(2662018JC051).
文摘The post-translational modification lysine 2-hydroxyisobutyrylation(K_(hib))plays an important role in gene transcription,metabolism,and enzymatic activity.K_(hib)sites have been identified in rice(Oryza sativa).However,the K_(hib)status of proteins in rice flowers during pathogen infection remains unclear.Here,we report a comprehensive identification of K_(hib)-modified proteins in rice flowers,and the changes in these proteins during infection with the fungal pathogen Ustilaginoidea virens.By using a tandem mass tag-based quantitative proteomics approach,we identified 2,891 K_(hib)sites on 964 proteins in rice flowers.Our data demonstrated that 2-hydroxyisobutyrylated proteins are involved in diverse biological processes.K_(hib)levels were substantially reduced upon infection with U.virens.Chromatin immunoprecipitation polymerase chain reaction(PCR)and reverse transcription quantitative PCR analyses revealed that histone K_(hib)is involved in the expression of disease-resistance genes.More importantly,most quantified sites on core histones H3 were downregulated upon U.virens infection.In addition,the histone deacetylases HDA705,HDA716,SRT1,and SRT2 are involved in the removal of K_(hib)marks in rice.HDA705 was further confirmed to negatively regulate rice disease resistance to pathogens U.virens,Magnaporthe oryzae,and Xanthomonas oryzae pv.oryzae(Xoo).Our data suggest that U.virens could modulate K_(hib)in rice flowers during infection.