Parkin,an E3 ubiquitin ligase,plays a role in maintaining mitochondrial homeostasis through targeting damaged mitochondria for mitophagy.Accumulating evidence suggests that the acetylation modification of the key mito...Parkin,an E3 ubiquitin ligase,plays a role in maintaining mitochondrial homeostasis through targeting damaged mitochondria for mitophagy.Accumulating evidence suggests that the acetylation modification of the key mitophagy machinery influences mitophagy level,but the underlying mechanism is poorly understood.Here,our study demonstrated that inhibition of histone deacetylase(HDAC)by treatment of HDACis activates mitophagy through mediating Parkin acetylation,leading to inhibition of cervical cancer cell proliferation.Bioinformatics analysis shows that Parkin expression is inversely correlated with HDAC2 expression in human cervical cancer,indicating the low acetylation level of Parkin.Using mass spectrometry,Parkin is identified to interact with two upstream molecules,acetylase acetyl-Co A acetyltransferase 1(ACAT1)and deacetylase HDAC2.Under treatment of suberoylanilide hydroxamic acid(SAHA),Parkin is acetylated at lysine residues 129,220 and 349,located in different domains of Parkin protein.In in vitro experiments,combined mutation of Parkin largely attenuate the interaction of Parkin with PTEN induced putative kinase 1(PINK1)and the function of Parkin in mitophagy induction and tumor suppression.In tumor xenografts,the expression of mutant Parkin impairs the tumor suppressive effect of Parkin and decreases the anticancer activity of SAHA.Our results reveal an acetylation-dependent regulatory mechanism governing Parkin in mitophagy and cervical carcinogenesis,which offers a new mitophagy modulation strategy for cancer therapy.展开更多
The mucosal barrier is crucial for intestinal homeostasis,and goblet cells are essential for maintaining the mucosal barrier integrity.The proviral integration site for Moloney murine leukemia virus-1(PIM1)kinase regu...The mucosal barrier is crucial for intestinal homeostasis,and goblet cells are essential for maintaining the mucosal barrier integrity.The proviral integration site for Moloney murine leukemia virus-1(PIM1)kinase regulates multiple cellular functions,but its role in intestinal homeostasis during colitis is unknown.Here,we demonstrate that PIM1 is prominently elevated in the colonic epithelia of both ulcerative colitis patients and murine models,in the presence of intestinal microbiota.Epithelial PIM1 leads to decreased goblet cells,thus impairing resistance to colitis and colitis-associated colorectal cancer(CAC)in mice.Mechanistically,PIM1 modulates goblet cell differentiation through the Wnt and Notch signaling pathways.Interestingly,PIM1 interacts with histone deacetylase 2(HDAC2)and downregulates its level via phosphorylation,thereby altering the epigenetic profiles of Wnt signaling pathway genes.Collectively,these findings investigate the unknown function of the PIM1-HDAC2 axis in goblet cell differentiation and ulcerative colitis/CAC pathogenesis,which points to the potential for PIM1-targeted therapies of ulcerative colitis and CAC.展开更多
目的:探讨组蛋白去乙酰化酶2(HDAC2)在生理浓度的糖皮质激素(GCs)抗炎效应中的作用。方法:脂多糖(LPS)(20 mg/L)孵育腹腔巨噬细胞诱导炎症反应。ELISA法测定细胞培养上清中炎症因子TNF-α和IL-1β水平;Western blot及Trans AM NF-κB p6...目的:探讨组蛋白去乙酰化酶2(HDAC2)在生理浓度的糖皮质激素(GCs)抗炎效应中的作用。方法:脂多糖(LPS)(20 mg/L)孵育腹腔巨噬细胞诱导炎症反应。ELISA法测定细胞培养上清中炎症因子TNF-α和IL-1β水平;Western blot及Trans AM NF-κB p65试剂盒检测NF-κB核转位和NF-κB与DNA的结合活性。HDAC阻断剂TSA或HDAC2 si RNA用于观察抑制HDAC2活性后对生理浓度的氢化可的松抑制LPS诱导的炎症反应及NF-κB活化的影响。结果 :LPS浓度依赖性的刺激TNF-α、IL-1β的产生,生理浓度的氢化可的松对此有明显的抑制作用,而预先给予TSA或HDAC2 si RNA,明显减弱氢化可的松对LPS诱导的TNF-α和IL-1β释放的抑制作用。进一步研究显示,LPS明显诱导NF-κB的核转位及与DNA的结合。生理浓度的氢化可的松减弱LPS诱导的p65/DNA结合从而抑制NF-κB的活化过程。预先给予TSA或HDAC2si RNA则可以阻断氢化可的松对NF-κB活化的抑制作用。结论 :HDAC2参与了生理浓度的GCs的抗炎效应,其机制与HDAC2介导的NF-κB转录活性的抑制有关。展开更多
Objective.Chronic stress(CS)-induced abnormal metabolism and other subsequent aspects of abnormality are threatening human health.Little is known regarding whether and how protein post-translational-modifications(PTMs...Objective.Chronic stress(CS)-induced abnormal metabolism and other subsequent aspects of abnormality are threatening human health.Little is known regarding whether and how protein post-translational-modifications(PTMs)correlate with abnormal metabolism under CS.The aim of this study was to address this issue and also identify novel key protein PTM.Methods.First,we screened which pan-PTM had significant change between control and CS female mice and whether clinical CS females had similar pan-PTM change.Second,we performed quantitative PTM-omics and metabolomics to verify the correlation between abnormal protein PTMs and atypical metabolism.Third,we performed quantitative phospho-omics to identify the key PTM-regulating enzyme and investigate the interaction between PTM protein and PTM-regulating enzyme.Fourth,we attempted to rectify the abnormal metabolism by correcting the activity of the PTM-regulating enzyme.Finally,we examined whether the selected key protein was also correlated with stress scores and atypical metabolism in clinical women.Results.We initially found that multiple tissues of CS female mice have downregulated pan-crotonylation,and verified that the plasma of clinical CS females also had downregulated pan-crotonylation.Then we determined that ATP5O-K51 crotonylation decreased the most and also caused gross ATP5O decrement,whereas the plasma of CS mice had downregulated phospholipids.Next,downregulating ATP5O crotonylation partially recapitulated the downregulated phospholipid metabolism in CS mice.Next,we verified that HDAC2-S424 phosphorylation determined its decrotonylation activity on ATP5O-K51.Furthermore,correcting HDAC2 hyper-phosphorylation recovered the gross ATP5O level and partially rescued the downregulated phospholipid metabolism in CS mice.Finally,the ATP5O level was also significantly iower and correlated with high stress scores and downregulated phospholipid metabolism in clinical female plasma.Conclusion.This study discovered a novel PTM mechanism involving two distinct types of PT展开更多
基金supported by research grants from Zhejiang Provincial Natural Science Foundation(LR18H160002,China)National Natural Science Foundation of China(32070740)+3 种基金Zhejiang Provincial Program in Medicine and Health Sciences and Technology(2018KY010)Zhejiang Provincial Outstanding Talent Project of Ten Thousand Talents ProgramZhejiang Provincial Qianjiang Talents ProgramZhejiang Provincial High-Level Innovative Health Talents Program to Dr.Jianbin Zhang。
文摘Parkin,an E3 ubiquitin ligase,plays a role in maintaining mitochondrial homeostasis through targeting damaged mitochondria for mitophagy.Accumulating evidence suggests that the acetylation modification of the key mitophagy machinery influences mitophagy level,but the underlying mechanism is poorly understood.Here,our study demonstrated that inhibition of histone deacetylase(HDAC)by treatment of HDACis activates mitophagy through mediating Parkin acetylation,leading to inhibition of cervical cancer cell proliferation.Bioinformatics analysis shows that Parkin expression is inversely correlated with HDAC2 expression in human cervical cancer,indicating the low acetylation level of Parkin.Using mass spectrometry,Parkin is identified to interact with two upstream molecules,acetylase acetyl-Co A acetyltransferase 1(ACAT1)and deacetylase HDAC2.Under treatment of suberoylanilide hydroxamic acid(SAHA),Parkin is acetylated at lysine residues 129,220 and 349,located in different domains of Parkin protein.In in vitro experiments,combined mutation of Parkin largely attenuate the interaction of Parkin with PTEN induced putative kinase 1(PINK1)and the function of Parkin in mitophagy induction and tumor suppression.In tumor xenografts,the expression of mutant Parkin impairs the tumor suppressive effect of Parkin and decreases the anticancer activity of SAHA.Our results reveal an acetylation-dependent regulatory mechanism governing Parkin in mitophagy and cervical carcinogenesis,which offers a new mitophagy modulation strategy for cancer therapy.
基金supported by grants from the National Natural Science Foundation of China(NSFC)Programs(32170186,82200618,82173199)Tianjin Science and Technology Commissioner Project(22JCZDJC00490,22JCQ NJC00540,China).
文摘The mucosal barrier is crucial for intestinal homeostasis,and goblet cells are essential for maintaining the mucosal barrier integrity.The proviral integration site for Moloney murine leukemia virus-1(PIM1)kinase regulates multiple cellular functions,but its role in intestinal homeostasis during colitis is unknown.Here,we demonstrate that PIM1 is prominently elevated in the colonic epithelia of both ulcerative colitis patients and murine models,in the presence of intestinal microbiota.Epithelial PIM1 leads to decreased goblet cells,thus impairing resistance to colitis and colitis-associated colorectal cancer(CAC)in mice.Mechanistically,PIM1 modulates goblet cell differentiation through the Wnt and Notch signaling pathways.Interestingly,PIM1 interacts with histone deacetylase 2(HDAC2)and downregulates its level via phosphorylation,thereby altering the epigenetic profiles of Wnt signaling pathway genes.Collectively,these findings investigate the unknown function of the PIM1-HDAC2 axis in goblet cell differentiation and ulcerative colitis/CAC pathogenesis,which points to the potential for PIM1-targeted therapies of ulcerative colitis and CAC.
文摘目的:探讨组蛋白去乙酰化酶2(HDAC2)在生理浓度的糖皮质激素(GCs)抗炎效应中的作用。方法:脂多糖(LPS)(20 mg/L)孵育腹腔巨噬细胞诱导炎症反应。ELISA法测定细胞培养上清中炎症因子TNF-α和IL-1β水平;Western blot及Trans AM NF-κB p65试剂盒检测NF-κB核转位和NF-κB与DNA的结合活性。HDAC阻断剂TSA或HDAC2 si RNA用于观察抑制HDAC2活性后对生理浓度的氢化可的松抑制LPS诱导的炎症反应及NF-κB活化的影响。结果 :LPS浓度依赖性的刺激TNF-α、IL-1β的产生,生理浓度的氢化可的松对此有明显的抑制作用,而预先给予TSA或HDAC2 si RNA,明显减弱氢化可的松对LPS诱导的TNF-α和IL-1β释放的抑制作用。进一步研究显示,LPS明显诱导NF-κB的核转位及与DNA的结合。生理浓度的氢化可的松减弱LPS诱导的p65/DNA结合从而抑制NF-κB的活化过程。预先给予TSA或HDAC2si RNA则可以阻断氢化可的松对NF-κB活化的抑制作用。结论 :HDAC2参与了生理浓度的GCs的抗炎效应,其机制与HDAC2介导的NF-κB转录活性的抑制有关。
基金supported by the General Program of the National Natural Science Foundation of China(Grant No:32070840)Dong Zhang and(Grant No:81571403)+2 种基金Xiang Ma,Nanjing Medical Science and Technology Development Project(Grant No:YKK18112)Jing Sun,the Independent Project of State Key Lab of Reproductive Medicine(Grant No:SKLRM-2021B6)Dong Zhang,and the Natural Science Foundation of Jiangsu Province(Grant No:BK20201355)to Dong Zhang.
文摘Objective.Chronic stress(CS)-induced abnormal metabolism and other subsequent aspects of abnormality are threatening human health.Little is known regarding whether and how protein post-translational-modifications(PTMs)correlate with abnormal metabolism under CS.The aim of this study was to address this issue and also identify novel key protein PTM.Methods.First,we screened which pan-PTM had significant change between control and CS female mice and whether clinical CS females had similar pan-PTM change.Second,we performed quantitative PTM-omics and metabolomics to verify the correlation between abnormal protein PTMs and atypical metabolism.Third,we performed quantitative phospho-omics to identify the key PTM-regulating enzyme and investigate the interaction between PTM protein and PTM-regulating enzyme.Fourth,we attempted to rectify the abnormal metabolism by correcting the activity of the PTM-regulating enzyme.Finally,we examined whether the selected key protein was also correlated with stress scores and atypical metabolism in clinical women.Results.We initially found that multiple tissues of CS female mice have downregulated pan-crotonylation,and verified that the plasma of clinical CS females also had downregulated pan-crotonylation.Then we determined that ATP5O-K51 crotonylation decreased the most and also caused gross ATP5O decrement,whereas the plasma of CS mice had downregulated phospholipids.Next,downregulating ATP5O crotonylation partially recapitulated the downregulated phospholipid metabolism in CS mice.Next,we verified that HDAC2-S424 phosphorylation determined its decrotonylation activity on ATP5O-K51.Furthermore,correcting HDAC2 hyper-phosphorylation recovered the gross ATP5O level and partially rescued the downregulated phospholipid metabolism in CS mice.Finally,the ATP5O level was also significantly iower and correlated with high stress scores and downregulated phospholipid metabolism in clinical female plasma.Conclusion.This study discovered a novel PTM mechanism involving two distinct types of PT
