Background Goat milk is increasingly recognized for high digestibility and a distinctive compositional profile.Protein acetylation,an important post-translational modification,regulates biosynthetic and metabolic path...Background Goat milk is increasingly recognized for high digestibility and a distinctive compositional profile.Protein acetylation,an important post-translational modification,regulates biosynthetic and metabolic pathways.This study aimed to identify critical acetylated proteins and specific modification sites involved in milk production and component synthesis in dairy goats,thereby elucidating the molecular mechanisms of lactation.We performed a comparative TMT-based acetylomic and proteomic analysis of mammary tissues from Saanen dairy goats during peak lactation and the dry period using LC–MS/MS.A candidate acetylation site was further investigated in goat mammary epithelial cells(GMECs)through site-directed mutagenesis and lipid metabolic assays,establishing functional links between acetylation and mammary lipid metabolism and providing a foundation for molecular strategies to improve milk quality and yield.Results We established a comprehensive mammary acetylome,identifying 862 significantly acetylated proteins and 2,028 modification sites across the two physiological phases.Differentially acetylated proteins were predominantly localized to the cytoplasm(39.98%).From these,54 key acetylated proteins,including MTOR,BCAT2,QARS1,GOT1,GOT2,BDH1,ACSS1,STAT5B,FABP5,and GPAM were prioritized as candidates involved in milk protein synthesis,milk fat synthesis,lactose synthesis,and other lactation-related processes.Among them,β-hydroxybutyrate dehydrogenase 1(BDH1)acetylation was characterized in detail.Members of the HDAC family were identified as primary regulators mediating BDH1 deacetylation.BDH1 acetylation promoted lipid droplet formation and triglyceride synthesis in GMECs.At the transcriptional level,BDH1 acetylation upregulated LXRα,ACSL1 and SCD1,whereas deacetylation downregulated SCD1,FASN,and ACSL1.Notably,BDH1 acetylation/deacetylation significantly reduced SREBP1 expression,linking this modification to coordinated control of lipogenic gene networks.Conclusions This study established,for the first time,the comprehensive acetylome of mammary gland tissues in dairy goats,revealing a substantial number of differentially acetylated proteins and modification sites.We demonstrate that acetylation of BDH1 regulated by HDACs promotes lipid droplet biogenesis and triglyceride synthesis in GMECs through transcriptional modulation of key lipogenic genes and suppression of SREBP1.These findings provide mechanistic insights into the post-translational regulation of mammary lipid metabolism and offer molecular targets for future genetic and nutritional strategies aimed at enhancing milk quality and yield in dairy goats.展开更多
Modern technological lifestyles promote allergic diseases,especially food allergies.The underlying molecular mechanisms remain to be uncovered.Protein acetylation is one of the most important post-translational modifi...Modern technological lifestyles promote allergic diseases,especially food allergies.The underlying molecular mechanisms remain to be uncovered.Protein acetylation is one of the most important post-translational modifications,and it is involved in regulating multiple body metabolic processes.This study aimed to clarify the effects of a high-fat diet(HFD)on allergy risk and the underlying mechanisms.Four-week-old male C57 BL/6 J mice were randomly divided into two groups and fed a normal fat diet(NFD)or HFD for 24 weeks.Then,serum lipids were measured,and skeletal muscle was collected for acetylome analysis.Compared with the findings in the NFD group,HFD-fed mice were obese and hyperlipidemic.Acetylome analysis also revealed 32 differentially expressed proteins between the HFD and NFD groups.Among these,eight acetylated proteins were upregulated in the HFD group.In addition,13 and 11 proteins were acetylated only in the HFD group and NFD group,respectively.These proteins were mainly involved in regulating energy metabolism and mitochondrial function.This study provides information regarding the underlying molecular mechanisms by which HFD promotes allergy.展开更多
Lysine acetylation,a reversible and dynamic post-translational modification,plays pivotal roles in regulating many biological processes in plants.However,the information of lysine acetylation during flower development...Lysine acetylation,a reversible and dynamic post-translational modification,plays pivotal roles in regulating many biological processes in plants.However,the information of lysine acetylation during flower development of woody angiosperm remains unclear.Herein,we identified a total of 667 lysine acetylation sites in 467 proteins in flower buds of Catalpa Bungei.The acetylated proteins were mainly involved in the pathways of carbon metabolism,glycometabolism and oxidative phosphorylation.Using functional enrichment analysis,61 and 13 acetylated proteins were involved in the glycometabolism and oxidative phosphorylation pathways,respectively.This suggests that lysine acetylated proteins play critical roles in the energy metabolism pathways.Accordingly,the protein–protein interactions were also highly enriched in the carbon metabolic pathway.Integrative analysis of lysine acetylome and transcriptome showed that transcript level changes of seven key lysine-acetylated proteins/genes involved in energy metabolism were significant different during floral transition.Importantly,compared with the normal flowering variety,the expression levels of phosphoribulokinase,phosphoenolpyruvate carboxykinase,ATP synthase and pyruvate dehydrogenase complex genes were significantly up-regulated at the stages of vegetative and transition buds during early flowering of C.bungei.These data represent the first lysine acetylome in the C.bungei flowers,and can serve as an essential basis for exploring the function of lysine acetylated proteins in the flower bud development of plants.展开更多
Post-translational modifications(PTMs)regulate the activity and functionality of RELA,but their role in the pathogenesis of liver fibrosis is unclear.This study was performed to understand the regulation mechanism of ...Post-translational modifications(PTMs)regulate the activity and functionality of RELA,but their role in the pathogenesis of liver fibrosis is unclear.This study was performed to understand the regulation mechanism of acetylation of RELA on liver inflammation and fibrosis in a model animal of innate glucose intolerance,largemouth bass,and to provide a potential target and biomarker for liver fibrosis therapy.We found that the acetylation of total proteins and RELA was significantly reduced in fibrotic livers of largemouth bass induced by a high-carbohydrate and high-fat diet(HCHFD)and CCL4 challenge.Furthermore,quantitative acetylome data showed that the K119 site of RELA was deacetylated in fibrotic livers compared to healthy controls.Subsequently,we reveal a new mechanism that SIRT7 deacetylates RELA at the K119 site in largemouth bass.RELA K119 deacetylation enhances RELA transcriptional activity by increasing its DNA-binding activity,and facilitates nuclear translocation of RELA,resulting in the overwhelming release of proinflammatory factors,and subsequently enhancing liver inflammation and fibrosis.Pharmacological inhibition of SIRT7 using a specific inhibitor restores the decreased acetylation of RELA in vivo and in vitro,and reduces the transcriptional activity,nuclear localization of RELA and the expression of its target genes,which ultimately attenuates liver inflammation and fibrosis.These findings uncover a novel mechanism underlying liver fibrosis involving SIRT7-mediated deacetylation of RELA to activate the proinflammatory gene program,and thus provide important insights and biomarkers into the effective strategies for limiting liver inflammation and fibrosis.展开更多
Lysine acetylation, an evolutionarily conserved post-translational protein modification, is reversibly catalyzed by lysine acetyltransferases and lysine deacetylases. Lysine acetylation, which was first discovered on ...Lysine acetylation, an evolutionarily conserved post-translational protein modification, is reversibly catalyzed by lysine acetyltransferases and lysine deacetylases. Lysine acetylation, which was first discovered on histones, mainly functions to configure the structure of chromatin and regulate gene transcriptional activity. Over the past decade, with advances in high-resolution mass spectrometry, a vast and growing number of non-histone proteins modified by acetylation in various plant species have been identified.Lysine acetylation of non-histone proteins is widely involved in regulating biological processes in plants such as photosynthesis, energy metabolism, hormone signal transduction and stress responses. Moreover, in plants, lysine acetylation plays crucial roles in regulating enzyme activity,protein stability, protein interaction and subcellular localization. This review summarizes recent progress in our understanding of the biological functions and mechanisms of non-histone protein acetylation in plants. Research prospects in this field are also noted.展开更多
Streptomycetes possess numerous gene clusters and the potential to produce a large amount of natural products.Histone deacetylase(HDAC)inhibitors play an important role in the regulation of histone modifications in fu...Streptomycetes possess numerous gene clusters and the potential to produce a large amount of natural products.Histone deacetylase(HDAC)inhibitors play an important role in the regulation of histone modifications in fungi,but their roles in prokaryotes remain poorly understood.Here,we investigated the global effects of the HDAC inhibitor,sodium butyrate(SB),on marine-derived Streptomyces olivaceus FXJ 8.021,particularly focusing on the activation of secondary metabolite biosynthesis.The antiSMASH analysis revealed 33 secondary metabolite biosynthetic gene clusters(BGCs)in strain FXJ 8.021,among which the silent lobophorin BGC was activated by SB.Transcriptomic data showed that the expression of genes involved in lobophorin biosynthesis(ge00097–ge00139)and CoA-ester formation(e.g.,ge02824),as well as the glycolysis/gluconeogenesis pathway(e.g.,ge01661),was significantly up-regulated in the presence of SB.Intracellular CoA-ester analysis confirmed that SB triggered the biosynthesis of CoA-ester,thereby increasing the precursor supply for lobophorin biosynthesis.Further acetylomic analysis revealed that the acetylation levels on 218 sites of 190 proteins were up-regulated and those on 411 sites of 310 proteins were down-regulated.These acetylated proteins were particularly enriched in transcriptional and translational machinery components(e.g.,elongation factor GE04399),and their correlations with the proteins involved in lobophorin biosynthesis were established by protein–protein interaction network analysis,suggesting that SB might function via a complex hierarchical.展开更多
基金supported by the National Key Research and Development Program of China(2022YFF1000102)Xi’an Agricultural Technology Research General Project(24NYGG0025)the National Natural Science Foundation of China(31702098)。
文摘Background Goat milk is increasingly recognized for high digestibility and a distinctive compositional profile.Protein acetylation,an important post-translational modification,regulates biosynthetic and metabolic pathways.This study aimed to identify critical acetylated proteins and specific modification sites involved in milk production and component synthesis in dairy goats,thereby elucidating the molecular mechanisms of lactation.We performed a comparative TMT-based acetylomic and proteomic analysis of mammary tissues from Saanen dairy goats during peak lactation and the dry period using LC–MS/MS.A candidate acetylation site was further investigated in goat mammary epithelial cells(GMECs)through site-directed mutagenesis and lipid metabolic assays,establishing functional links between acetylation and mammary lipid metabolism and providing a foundation for molecular strategies to improve milk quality and yield.Results We established a comprehensive mammary acetylome,identifying 862 significantly acetylated proteins and 2,028 modification sites across the two physiological phases.Differentially acetylated proteins were predominantly localized to the cytoplasm(39.98%).From these,54 key acetylated proteins,including MTOR,BCAT2,QARS1,GOT1,GOT2,BDH1,ACSS1,STAT5B,FABP5,and GPAM were prioritized as candidates involved in milk protein synthesis,milk fat synthesis,lactose synthesis,and other lactation-related processes.Among them,β-hydroxybutyrate dehydrogenase 1(BDH1)acetylation was characterized in detail.Members of the HDAC family were identified as primary regulators mediating BDH1 deacetylation.BDH1 acetylation promoted lipid droplet formation and triglyceride synthesis in GMECs.At the transcriptional level,BDH1 acetylation upregulated LXRα,ACSL1 and SCD1,whereas deacetylation downregulated SCD1,FASN,and ACSL1.Notably,BDH1 acetylation/deacetylation significantly reduced SREBP1 expression,linking this modification to coordinated control of lipogenic gene networks.Conclusions This study established,for the first time,the comprehensive acetylome of mammary gland tissues in dairy goats,revealing a substantial number of differentially acetylated proteins and modification sites.We demonstrate that acetylation of BDH1 regulated by HDACs promotes lipid droplet biogenesis and triglyceride synthesis in GMECs through transcriptional modulation of key lipogenic genes and suppression of SREBP1.These findings provide mechanistic insights into the post-translational regulation of mammary lipid metabolism and offer molecular targets for future genetic and nutritional strategies aimed at enhancing milk quality and yield in dairy goats.
基金funded by the 111 project from the Education Ministry of China(B18053)。
文摘Modern technological lifestyles promote allergic diseases,especially food allergies.The underlying molecular mechanisms remain to be uncovered.Protein acetylation is one of the most important post-translational modifications,and it is involved in regulating multiple body metabolic processes.This study aimed to clarify the effects of a high-fat diet(HFD)on allergy risk and the underlying mechanisms.Four-week-old male C57 BL/6 J mice were randomly divided into two groups and fed a normal fat diet(NFD)or HFD for 24 weeks.Then,serum lipids were measured,and skeletal muscle was collected for acetylome analysis.Compared with the findings in the NFD group,HFD-fed mice were obese and hyperlipidemic.Acetylome analysis also revealed 32 differentially expressed proteins between the HFD and NFD groups.Among these,eight acetylated proteins were upregulated in the HFD group.In addition,13 and 11 proteins were acetylated only in the HFD group and NFD group,respectively.These proteins were mainly involved in regulating energy metabolism and mitochondrial function.This study provides information regarding the underlying molecular mechanisms by which HFD promotes allergy.
基金This work was financially supported by the Open Fund of State Key Laboratory of Tree Genetics and Breeding(Chinese Academy of Forestry)(No.TGB2019005)Fundamental Research Funds for the Central Universities(XDJK2020B058).The funders had no role in study design,data collection and analysis,decision to publish,or preparation of the manuscript.
文摘Lysine acetylation,a reversible and dynamic post-translational modification,plays pivotal roles in regulating many biological processes in plants.However,the information of lysine acetylation during flower development of woody angiosperm remains unclear.Herein,we identified a total of 667 lysine acetylation sites in 467 proteins in flower buds of Catalpa Bungei.The acetylated proteins were mainly involved in the pathways of carbon metabolism,glycometabolism and oxidative phosphorylation.Using functional enrichment analysis,61 and 13 acetylated proteins were involved in the glycometabolism and oxidative phosphorylation pathways,respectively.This suggests that lysine acetylated proteins play critical roles in the energy metabolism pathways.Accordingly,the protein–protein interactions were also highly enriched in the carbon metabolic pathway.Integrative analysis of lysine acetylome and transcriptome showed that transcript level changes of seven key lysine-acetylated proteins/genes involved in energy metabolism were significant different during floral transition.Importantly,compared with the normal flowering variety,the expression levels of phosphoribulokinase,phosphoenolpyruvate carboxykinase,ATP synthase and pyruvate dehydrogenase complex genes were significantly up-regulated at the stages of vegetative and transition buds during early flowering of C.bungei.These data represent the first lysine acetylome in the C.bungei flowers,and can serve as an essential basis for exploring the function of lysine acetylated proteins in the flower bud development of plants.
基金supported by National Natural Sci-ence Foundation of China(32273141,32202954,32172981)Beijing Natural Science Foundation(6232038).
文摘Post-translational modifications(PTMs)regulate the activity and functionality of RELA,but their role in the pathogenesis of liver fibrosis is unclear.This study was performed to understand the regulation mechanism of acetylation of RELA on liver inflammation and fibrosis in a model animal of innate glucose intolerance,largemouth bass,and to provide a potential target and biomarker for liver fibrosis therapy.We found that the acetylation of total proteins and RELA was significantly reduced in fibrotic livers of largemouth bass induced by a high-carbohydrate and high-fat diet(HCHFD)and CCL4 challenge.Furthermore,quantitative acetylome data showed that the K119 site of RELA was deacetylated in fibrotic livers compared to healthy controls.Subsequently,we reveal a new mechanism that SIRT7 deacetylates RELA at the K119 site in largemouth bass.RELA K119 deacetylation enhances RELA transcriptional activity by increasing its DNA-binding activity,and facilitates nuclear translocation of RELA,resulting in the overwhelming release of proinflammatory factors,and subsequently enhancing liver inflammation and fibrosis.Pharmacological inhibition of SIRT7 using a specific inhibitor restores the decreased acetylation of RELA in vivo and in vitro,and reduces the transcriptional activity,nuclear localization of RELA and the expression of its target genes,which ultimately attenuates liver inflammation and fibrosis.These findings uncover a novel mechanism underlying liver fibrosis involving SIRT7-mediated deacetylation of RELA to activate the proinflammatory gene program,and thus provide important insights and biomarkers into the effective strategies for limiting liver inflammation and fibrosis.
基金supported by grants from the National Natural Science Foundation of China (No. 32070551 and No. 32371326)the Youth Innovation Promotion Association, CAS (No. 201860)Key Research Program of Frontier Sciences, Chinese Academy of Sciences (ZDBS-LY-SM009)。
文摘Lysine acetylation, an evolutionarily conserved post-translational protein modification, is reversibly catalyzed by lysine acetyltransferases and lysine deacetylases. Lysine acetylation, which was first discovered on histones, mainly functions to configure the structure of chromatin and regulate gene transcriptional activity. Over the past decade, with advances in high-resolution mass spectrometry, a vast and growing number of non-histone proteins modified by acetylation in various plant species have been identified.Lysine acetylation of non-histone proteins is widely involved in regulating biological processes in plants such as photosynthesis, energy metabolism, hormone signal transduction and stress responses. Moreover, in plants, lysine acetylation plays crucial roles in regulating enzyme activity,protein stability, protein interaction and subcellular localization. This review summarizes recent progress in our understanding of the biological functions and mechanisms of non-histone protein acetylation in plants. Research prospects in this field are also noted.
基金supported by the National Key R&D Program of China(Grant No.2020YFA0907800)the National Natural Science Foundation of China(Grant Nos.32170043 and 82173720).
文摘Streptomycetes possess numerous gene clusters and the potential to produce a large amount of natural products.Histone deacetylase(HDAC)inhibitors play an important role in the regulation of histone modifications in fungi,but their roles in prokaryotes remain poorly understood.Here,we investigated the global effects of the HDAC inhibitor,sodium butyrate(SB),on marine-derived Streptomyces olivaceus FXJ 8.021,particularly focusing on the activation of secondary metabolite biosynthesis.The antiSMASH analysis revealed 33 secondary metabolite biosynthetic gene clusters(BGCs)in strain FXJ 8.021,among which the silent lobophorin BGC was activated by SB.Transcriptomic data showed that the expression of genes involved in lobophorin biosynthesis(ge00097–ge00139)and CoA-ester formation(e.g.,ge02824),as well as the glycolysis/gluconeogenesis pathway(e.g.,ge01661),was significantly up-regulated in the presence of SB.Intracellular CoA-ester analysis confirmed that SB triggered the biosynthesis of CoA-ester,thereby increasing the precursor supply for lobophorin biosynthesis.Further acetylomic analysis revealed that the acetylation levels on 218 sites of 190 proteins were up-regulated and those on 411 sites of 310 proteins were down-regulated.These acetylated proteins were particularly enriched in transcriptional and translational machinery components(e.g.,elongation factor GE04399),and their correlations with the proteins involved in lobophorin biosynthesis were established by protein–protein interaction network analysis,suggesting that SB might function via a complex hierarchical.
