AIM: To explore the effects of IκBα SUMOylation and NF-κB p65 deacetylation on NF-κB p65 activity induced by high glucose in cultured human lens epithelial cells(HLECs).METHODS: HLECs(SRA01/04) were cultured with ...AIM: To explore the effects of IκBα SUMOylation and NF-κB p65 deacetylation on NF-κB p65 activity induced by high glucose in cultured human lens epithelial cells(HLECs).METHODS: HLECs(SRA01/04) were cultured with 5.5, 25, and 50 mmol/L glucose media for 24 h, and with 50 mmol/L glucose media for 0, 12, and 24 h respectively. SUMO1 and SIRT1 expressions were detected by reverse transcriptionpolymerase chain reaction(RT-PCR) and Western blot(WB). IκBα and NF-κB p65 expressions were detected by WB. With NAC, DTT, MG132 or Resveratrol(RSV) treatment, SUMO1 and SIRT1 expressions were detected by WB. Protein expression localizations were examined by immunofluorescence and co-immunofluorescence. The effects of SUMO1 or SIRT1 overexpression, as well as MG132 and RSV, on the nuclear expression and activity of IκBα and NF-κB p65 were analyzed by immunoblot and dual luciferase reporter gene assay.RESULTS: SUMO1 and SIRT1 expressions were influenced by high glucose in mRNA and protein levels, which could be blocked by NAC or DTT. SUMO1 was down-regulated by using MG132, and SIRT1 was up-regulated under RSV treatment. IκBα nuclear expression was attenuated and NF-κB p65 was opposite under high glucose, while IκBα and NF-κB p65 location was transferred to the nucleus. SUMO1 or SIRT1 overexpression and MG132 or RSV treatment affected the nuclear expression and activity of IκBα and NF-κB p65 under high glucose condition.CONCLUSION: IκBα SUMOylation and NF-κB p65 deacetylation affect NF-κB p65 activity in cultured HLECs under high glucose, and presumably play a significant role in controlling diabetic cataract.展开更多
SRY-related HMG-box(Sox) transcription factors are known to regulate central nervous system development and are involved in several neurological diseases.Post-translational modification of Sox proteins is known to alt...SRY-related HMG-box(Sox) transcription factors are known to regulate central nervous system development and are involved in several neurological diseases.Post-translational modification of Sox proteins is known to alter their functions in the central nervous system.Among the different types of post-translational modification,small ubiquitin-like modifier(SUMO) modification of Sox proteins has been shown to modify their transcriptional activity.Here,we review the mechanisms of three Sox proteins in neuronal development and disease,along with their transcriptional changes under SUMOylation.Across three species,lysine is the conserved residue for SUMOylation.In Drosophila,SUMOylation of Sox N plays a repressive role in transcriptional activity,which impairs central nervous system development.However,de SUMOylation of Sox E and Sox11 plays neuroprotective roles,which promote neural crest precursor formation in Xenopus and retinal ganglion cell differentiation as well as axon regeneration in the rodent.We further discuss a potential translational therapy by SUMO site modification using AAV gene transduction and Clustered regularly interspaced short palindromic repeats-Cas9 technology.Understanding the underlying mechanisms of Sox SUMOylation,especially in the rodent system,may provide a therapeutic strategy to address issues associated with neuronal development and neurodegeneration.展开更多
Human parainfluenza virus type 3(HPIV3),a member of the Paramyxoviridae family,can cause lower respiratory disease in infants and young children.The phosphoprotein(P)of HPIV3 is an essential cofactor of the viral RNA-...Human parainfluenza virus type 3(HPIV3),a member of the Paramyxoviridae family,can cause lower respiratory disease in infants and young children.The phosphoprotein(P)of HPIV3 is an essential cofactor of the viral RNA-dependent RNA polymerase large protein(L).P connects nucleocapsid protein(N)with L to initiate genome transcription and replication.Sumoylation influences many important pathways of the target proteins,and many viral proteins are also themselves sumoylated.In this study,we found that the P of HPIV3 could be sumoylated,and mutation of K492 and K532 to arginine(PK492 R/K532 R)failed to be sumoylated within P,which enhances HPIV3 minigenome activity.Biochemical studies showed that PK492 R/K532 Rhad no effect on its interactions with N,formation of homo-tetramers and formation of inclusion bodies.Finally,we found that incorporation of K492 R/K532 R into a recombinant HPIV3(rHPIV3-PK492 R/K532 R)increased viral production in culture cells,suggesting that sumoylation attenuates functions of P and down-regulates viral replication.展开更多
This study aims to investigate the impact of hepatocyte nuclear factor 1β(HNF1b)on macrophage sortilin-mediated lipid metabolism and aortic atherosclerosis and explore the role of the flavone of Polygonatum odoratum(...This study aims to investigate the impact of hepatocyte nuclear factor 1β(HNF1b)on macrophage sortilin-mediated lipid metabolism and aortic atherosclerosis and explore the role of the flavone of Polygonatum odoratum(PAOA-flavone)-promoted small ubiquitin-related modifier(SUMO)modification in the atheroprotective efficacy of HNF1b.HNF1b was predicted to be a transcriptional regulator of sortilin expression via bioinformatics,dual-luciferase reporter gene assay,and chromatin immunoprecipitation.HNF1b overexpression decreased sortilin expression and cellular lipid contents in THP-1 macrophages,leading to a depression in atherosclerotic plaque formation in low-density lipoprotein(LDL)receptor-deficient(LDLR−/−)mice.Multiple SUMO1-modified sites were identified on the HNF1b protein and co-immunoprecipitation confirmed its SUMO1 modification.The SUMOylation of HNF1b protein enhanced the HNF1b-inhibited effect on sortilin expression and reduced lipid contents in macrophages.PAOA-flavone treatment promoted SUMO-activating enzyme subunit 1(SAE1)expression and SAE1-catalyzed SUMOylation of the HNF1b protein,which prevented sortilin-mediated lipid accumulation in macrophages and the formation of atherosclerotic plaques in apolipoprotein E-deficient(ApoE−/−)mice.Interference with SAE1 abrogated the improvement in lipid metabolism in macrophage cells and atheroprotective efficacy in vivo upon PAOA-flavone administration.In summary,HNF1b transcriptionally suppressed sortilin expression and macrophage lipid accumulation to inhibit aortic lipid deposition and the development of atherosclerosis.This anti-atherosclerotic effect was enhanced by PAOA-flavone-facilitated,SAE1-catalyzed SUMOylation of the HNF1b protein.展开更多
Recently, Khayachi et al.;showed that fragile X mental retardation protein （FMRP） is an active substrate of the small ubiquitin-like modifier （SUMO） pathway in neurons.FMRP SUMOylation is induced by the activation...Recently, Khayachi et al.;showed that fragile X mental retardation protein （FMRP） is an active substrate of the small ubiquitin-like modifier （SUMO） pathway in neurons.FMRP SUMOylation is induced by the activation of metabotropic glutamate receptors （mGlu5Rs）. FMRP展开更多
Background:Abnormal myocardial voltage-gated sodium channel 1.5(Nav1.5)expression and function cause lethal ventricular arrhythmias during myocardial ischemia–reperfusion(I/R).Protein inhibitor of activated STAT Y(PI...Background:Abnormal myocardial voltage-gated sodium channel 1.5(Nav1.5)expression and function cause lethal ventricular arrhythmias during myocardial ischemia–reperfusion(I/R).Protein inhibitor of activated STAT Y(PIASy)-mediated caveolin-3(Cav-3)small ubiquitin-related modifier(SUMO)modification affects Cav-3 binding to the Nav1.5.PIASy activity is increased after myocardial I/R,but it is unclear whether this is attributable to plasma membrane Nav1.5 downregulation and ventricular arrhythmias.Methods:Using recombinant adeno-associated virus subtype 9(AAV9),rat cardiac PIASy was silenced using intraventricular injection of PIASy short hairpin RNA(shRNA).After two weeks,rat hearts were subjected to I/R and electrocardiography was performed to assess malignant arrhythmias.Tissues from peri-infarct areas of the left ventricle were collected for molecular biological measurements.Results:PIASy was upregulated by I/R(P<0.01),with increased SUMO2/3 modification of Cav-3 and reduced membrane Nav1.5 density(P<0.01).AAV9-PIASy shRNA intraventricular injection into the rat heart down-regulated PIASy after I/R,at both mRNA and protein levels(P<0.05 vs.Scramble-shRNA+I/R group),decreased SUMO-modified Cav-3 levels,enhanced Cav-3 binding to Nav1.5,and prevented I/R-induced decrease of Nav1.5 and Cav-3co-localization in the intercalated disc and lateral membrane.PIASy silencing in rat hearts reduced I/R-induced fatal arrhythmias,which was reflected by a modest decrease in the duration of ventricular fibrillation(VF;P<0.05 vs.Scramble-shRNA+I/R group)and a significantly reduced arrhythmia score(P<0.01 vs.Scramble-shRNA+I/R group).The anti-arrhythmic effects of PIASy silencing were also evidenced by decreased episodes of ventricular tachycardia(VT),sustained VT and VF,especially at the time 5–10 min after ischemia(P<0.05 vs.Scramble-shRNA+IR group).Using in vitro human embryonic kidney 293 T(HEK293T)cells and isolated adult rat cardiomyocyte models exposed to hypoxia/reoxygenation(H/R),we confirmed that increased PIASy promoted Cav-3 modification by SUMO2/3 and Nav1.5/Cav-3 dissociation after H/R.Mutation of SUMO consensus lysine sites in Cav-3(K38R or K144R)altered the membrane expression levels of Nav1.5 and Cav-3 before and after H/R in HEK293T cells.Conclusions:I/R-induced cardiac PIASy activation increased Cav-3 SUMOylation by SUMO2/3 and dysregulated Nav1.5-related ventricular arrhythmias.Cardiac-targeted PIASy silencing mediated Cav-3 deSUMOylation and partially prevented I/R-induced Nav1.5 downregulation in the plasma membrane of cardiomyocytes,and subsequent ventricular arrhythmias in rats.PIASy was identified as a potential therapeutic target for life-threatening arrhythmias in patients with ischemic heart diseases.展开更多
Objective This study elucidated the function and role of SUMOylation in type Ⅰ endometrial carcinoma.Methods Fifty type Ⅰ endometrial carcinoma cases and para-cancer tissue samples were collected.The expression leve...Objective This study elucidated the function and role of SUMOylation in type Ⅰ endometrial carcinoma.Methods Fifty type Ⅰ endometrial carcinoma cases and para-cancer tissue samples were collected.The expression levels of ubiquitin-conjugating enzyme E2I(Ube2 i,Ubc9) and small ubiquitin-like modifier 1(SUMO1)/sentrin-specific peptidase 1(SENP1) proteins were examined using immunohistochemistry and the correlation with clinicopathological parameters was analyzed.Results Ubc9 expression in type Ⅰ endometrial carcinoma tissues was significantly higher than that in the para-cancer tissues;in contrast,the expression of the SENP1 protein was markedly lower than that in the para-cancer tissues.Ubc9 and SENP1 expression levels were negatively correlated and were associated with tumor differentiation,but not age,depth of invasion,tumor stage,and lymph node metastasis.Conclusion SUMOylation modification plays a major role in the pathogenesis and development of type Ⅰ endometrial carcinoma.Thus,it could be a potential target for the treatment of endometrial cancer.展开更多
Dear Editor, In recent years, post-translational modifications （PTMs） by small ubiquitin-related modifiers （SUMOs） have emerged as an important regulatory mechanism for both cellular and viral processes （Ribet an...Dear Editor, In recent years, post-translational modifications （PTMs） by small ubiquitin-related modifiers （SUMOs） have emerged as an important regulatory mechanism for both cellular and viral processes （Ribet and Cossart, 2010）. Identifying the SUMOylation sites of the target protein is important to understand the molecular mechanism under- lying SUMO modification and virus-host interactions, as well as provide new insights into antiviral drug develop- ment （Wimmer and Schreiner, 2015）. Traditional site- directed mutagenesis for identifying viral protein SUMO- ylation sites lacks a specific aim and is laborious （McManus et al., 2016）. Recently, mass spectrometry （MS） has been employed as an accurate and sensitive tool to identify PTM sites, thereby greatly expanding the number of known SUMOylated proteins （Pedrioli et al., 2006）. However, during viral infection, SUMOylation is highly dynamic and SUMOylated viral proteins often have low abundance, which makes studying SUMOylation under natural conditions difficult.展开更多
Recently there have been exciting research advances in neuroprotective therapies for ischemic stroke. In the past, the search for neu- roprotective agents has been fraught with failure at the clinical trials stage due...Recently there have been exciting research advances in neuroprotective therapies for ischemic stroke. In the past, the search for neu- roprotective agents has been fraught with failure at the clinical trials stage due to numerous factors, including subject heterogeneity and improper therapeutic windows (Tymianski, 2017). Moreover, it is becoming clearer that the complex and evolving pathobiology of stroke requires multimodal therapeutic approaches capable of modulating the numerous axes that contribute to ischemia/reperfusion damage, rather than targeting a single axis (Bernstock et al., 2018a). With the success of recent endovascular thrombectomy (EVT) trials, it has been suggested that clinical trials of EVT with adjunct neuroprotection can overcome past difficulties and maximize the effect size by using imaging to reduce patient heterogeneity (i. e., selecting those with large vessel occlusions, small ischemic cores, and good collateral circulation), restoring perfusion using better EVT devices, and enrolling patients in the correct therapeutic window (i.e., when they still have salvageable brain tissue) (Tymianski, 2017). Considering the opportunity that this represents for new, better clinical trials of neuroprotective agents, the search is on for high-potential compounds that may be investigated in these future studies.展开更多
The centromere is a specialized domain that facilitates chromosome segregation during mitosis and serves as the site of kinetochore formation.KINETOCHORE NULL2(αKNL2)is essential for the recognition and loading of th...The centromere is a specialized domain that facilitates chromosome segregation during mitosis and serves as the site of kinetochore formation.KINETOCHORE NULL2(αKNL2)is essential for the recognition and loading of the centromeric histone H3 variant CENH3 at centromeres.A yeast two-hybrid screen forαKNL2 interactors identified components of the SUMOylation pathway.However,the role ofαKNL2 SUMOylation in Arabidopsis has not yet been determined.In this study,we demonstrated that the C-terminal region ofαKNL2(designatedαKNL2-C)interacts with small ubiquitin-like modifier 3(SUMO3)and ULP1d,as shown by bimolecular fluorescence complementation and co-immunoprecipitation assays.Bioinformatic and functional analyses ofαKNL2-C identified three SUMOylation sites and two SUMOinteracting motifs,which were shown to be critical for growth,fertility,and chromosome alignment.Of the three SUMOylation sites,Lys474 and Lys511 are the most critical for the centromeric localization ofαKNL2,underscoring the importance ofαKNL2 SUMOylation for its function.Additionally,both in vitro and in vivo assays showed thatαKNL2-C undergoes SUMOylation by SUMO1 or SUMO3.The Arabidopsis SUMO protease mutant ulp1d-2 exhibits a mild accumulation of SUMOylatedαKNL2.We further showed that SUMOylation ofαKNL2 promotes its binding to CENH3 and controls protein stability.Our findings demonstrate that C-terminal SUMOylation ofαKNL2 is crucial for its centromeric localization,interaction with CENH3,and kinetochore assembly,emphasizing the significance of post-translational modifications in chromosome segregation and cell division in plants.展开更多
SUMO PROTEASE RELATED TO FERTILITY 1(SPF1)and SPF2 are responsible for deSUMOylation of SUMO-conjugated protein substrates and for maintaining protein SUMOylation homeostasis.To date,the role of SUMO proteases in fatt...SUMO PROTEASE RELATED TO FERTILITY 1(SPF1)and SPF2 are responsible for deSUMOylation of SUMO-conjugated protein substrates and for maintaining protein SUMOylation homeostasis.To date,the role of SUMO proteases in fatty acid biosynthesis and lipid accumulation remains unclear.Here,we demonstrate that the Arabidopsis thaliana mutants spf1-1,spf2-1,and spf1-1 spf2-1 exhibit increased seed size and elevated seed oil content(SOC).We further show that SPF1 and SPF2 interact with WRINKLED1(WRI1),a master regulator of the transcriptional control of plant oil synthesis.Genetic analyses indicate that the spf1-1 wri1-3 and spf2-1 wri1-3 double mutants,as well as the spf1-1 spf2-1 wri1-3 triple mutant,phenocopy wri1-3 and display severe seed shriveling,indicating that SPF1 and SPF2 act upstream of WRI1.WRI1 was identified as a SUMO1 substrate with two conserved SUMOylation sites,lysine 257(K257)and K266,in cruciferous plants,with K257 acting as the dominant site required for seed oil synthesis.SUMOylation enhances WRI1 stability,whereas SPF1and SPF2mediated deSUMOylation promotes WRI1 degradation.In spf1-1,spf2-1,and spf1-1 spf2-1 mutants,the abundance of SUMOylated WRI1 increases during seed development and correlates with elevated seed oil accumulation.Together,these results indicate that SPF1 and SPF2 negatively regulate oil synthesis by deSUMOylating WRI1,establishing a dynamic SUMOylation and deSUMOylation switch centered on the SPF1/SPF2-WRI1 module that fine-tunes seed development and oil synthesis.展开更多
Most cancers are currently incurable,partly due to abnormal post-translational modifications(PTMs).In this study,we initially used multiple myeloma(MM)as a working model and found that SUMOylation activating enzyme su...Most cancers are currently incurable,partly due to abnormal post-translational modifications(PTMs).In this study,we initially used multiple myeloma(MM)as a working model and found that SUMOylation activating enzyme subunit 1(SAE1)promotes the malignancy of MM.Through proteome microarray analysis,SAE1 was identified as a potential target for bioactive colcemid or its derivative colchicine.Elevated levels of SAE1 were associated with poor clinical survival and increased MM proliferation in vitro and in vivo.Additionally,SAE1 directly SUMOylated and upregulated the total protein expression of p27,leading to LLPS-mediated nuclear export of p27.Our study also demonstrated the involvement of SAE1 in other types of cancer cells,and provided the first monomer crystal structure of SAE1 and its key binding model with colchicine.Colchicine also showed promising results in the Patient-Derived Tumor Xenograft(PDX)model.Furthermore,a controlled clinical trial with 56 MM patients demonstrated the clinical efficacy of colchicine.Our findings reveal a novel mechanism by which tumor cells evade p27-induced cellular growth arrest through p27 SUMOylation-mediated nuclear export.SAE1 may serve as a promising therapeutic target,and colchicine may be a potential treatment option for multiple types of cancer in clinical settings.展开更多
SUMOylation is a reversible posttranslational modification that plays a crucial role in various essential biological processes within eukaryotic cells.This reaction involves the transfer of small ubiquitin-like modifi...SUMOylation is a reversible posttranslational modification that plays a crucial role in various essential biological processes within eukaryotic cells.This reaction involves the transfer of small ubiquitin-like modifier(SUMO)molecules to lysine residues on target proteins,thereby regulating the stability,localization,and activity of these substrates.Despite the low amino acid sequence similarity between SUMO and ubiquitin,they share similar three-dimensional structures and are conjugated onto protein substrates through a comparable three-step catalytic reaction.展开更多
Chemo-/radioresistance of malignant tumors hampers cancer control and increases patient mortality.Efficient repair of damaged DNA is critical for the maintenance of genomic integrity and fidelity of genetic informatio...Chemo-/radioresistance of malignant tumors hampers cancer control and increases patient mortality.Efficient repair of damaged DNA is critical for the maintenance of genomic integrity and fidelity of genetic information.In reverse,increased DNA repair capability in cancer cells contributes to chemo-/radioresistance of malignant tumors.DNA double-strand break(DSB)is the most serious DNA damage and is also the principal molecular basis of radiotherapy.Upon DNA damage,the Ku80 is recruited and forms a critical DNA-PK complex at the DSB sites with Ku70 and the catalytic subunit(DNA-PKcs)to initiate DNA repair.How DNA-PK is assembled and activated is not fully understood.Based on the identification of radiation-reduced Ku80 K568 crotonylation through quantitative global lysine crotonylome analysis,we reveal that Ku80 K568 is crotonylated by p300-CBP-associated factor(PCAF).Upon DNA damage,the K568cr is decrotonylated by HDAC8(Histone deacetylase 8).Decrotonylation of K568cr empties this site for the subsequent SUMOylation of Ku80 by CBX4.The conversion of Ku80 from K568 crotonylation to SUMOylation facilitates the assembly of DNA-PK complex and autophosphorylation of DNA-PKcs S2056,consequently activating the DSB repair.Moreover,mutation disrupting the post-translational modification(PTM)of Ku80 K568 site sensitizes cancer cells to radiotherapy in tumorbearing nude mice models.This study elucidates the conversion model between two different forms of PTMs in the regulation of DNA-PK complex assembly and DSB repair,highlighting this model’s potential in controlling chemo-/radioresistance of malignant tumors,as well as expands the atlas of therapeutic targets.展开更多
Dear Editor,SUMOylation is a crucial form of post-translational modification in which small ubiquitin-like modifiers(SUMOs)are transferred to lysine residues of target proteins,thereby regulating their stability,local...Dear Editor,SUMOylation is a crucial form of post-translational modification in which small ubiquitin-like modifiers(SUMOs)are transferred to lysine residues of target proteins,thereby regulating their stability,localization,and activity.Proteomic analysis in Arabidopsis has revealed numerous proteins serving as SUMOylation substrates,which play crucial roles in development and stress responses(Elrouby and Coupland,2010).展开更多
The growing interest in post-translational protein modification,particularly in SU-MOylation,is driven by its crucial role in cell cycle regulation.SUMOylation affects various cell cycle regulators,including oncogenes...The growing interest in post-translational protein modification,particularly in SU-MOylation,is driven by its crucial role in cell cycle regulation.SUMOylation affects various cell cycle regulators,including oncogenes,suggesting its relevance in cancer.SUMO E3 ligases are pivotal in this process,exhibiting diverse functionalities through structural domains and sub-cellular localizations.A less-explored SUMO E3 ligase,RANBP2,a component of the vertebrate nuclear pore complex,emerges as a central player in cellular cycle processes,as well as in tumorigenesis.The current studies illuminate the importance of RANBP2 and underscore the need for more extensive studies to validate its clinical applicability in neoplastic interventions.Our review elucidates the significance of RANBP2 across various types of malignancies.Addi-tionally,it delves into exploring RANBP2 as a prospective therapeutic target for cancer treat-ment,offering insights into the avenues that scholars should pursue in their subsequent research endeavors.Thus,further investigation into RANBP2’s role in solid tumorigenesis is eagerly awaited.展开更多
基金Supported by the National Natural Science Foundation of China(No.81170836, No.81570838)
文摘AIM: To explore the effects of IκBα SUMOylation and NF-κB p65 deacetylation on NF-κB p65 activity induced by high glucose in cultured human lens epithelial cells(HLECs).METHODS: HLECs(SRA01/04) were cultured with 5.5, 25, and 50 mmol/L glucose media for 24 h, and with 50 mmol/L glucose media for 0, 12, and 24 h respectively. SUMO1 and SIRT1 expressions were detected by reverse transcriptionpolymerase chain reaction(RT-PCR) and Western blot(WB). IκBα and NF-κB p65 expressions were detected by WB. With NAC, DTT, MG132 or Resveratrol(RSV) treatment, SUMO1 and SIRT1 expressions were detected by WB. Protein expression localizations were examined by immunofluorescence and co-immunofluorescence. The effects of SUMO1 or SIRT1 overexpression, as well as MG132 and RSV, on the nuclear expression and activity of IκBα and NF-κB p65 were analyzed by immunoblot and dual luciferase reporter gene assay.RESULTS: SUMO1 and SIRT1 expressions were influenced by high glucose in mRNA and protein levels, which could be blocked by NAC or DTT. SUMO1 was down-regulated by using MG132, and SIRT1 was up-regulated under RSV treatment. IκBα nuclear expression was attenuated and NF-κB p65 was opposite under high glucose, while IκBα and NF-κB p65 location was transferred to the nucleus. SUMO1 or SIRT1 overexpression and MG132 or RSV treatment affected the nuclear expression and activity of IκBα and NF-κB p65 under high glucose condition.CONCLUSION: IκBα SUMOylation and NF-κB p65 deacetylation affect NF-κB p65 activity in cultured HLECs under high glucose, and presumably play a significant role in controlling diabetic cataract.
基金supported by NIH CORE Grant P30 EY08098 to the Department of Ophthalmology,University of Pittsburgh,the Eye and Ear Foundation of Pittsburgh (to KCC)。
文摘SRY-related HMG-box(Sox) transcription factors are known to regulate central nervous system development and are involved in several neurological diseases.Post-translational modification of Sox proteins is known to alter their functions in the central nervous system.Among the different types of post-translational modification,small ubiquitin-like modifier(SUMO) modification of Sox proteins has been shown to modify their transcriptional activity.Here,we review the mechanisms of three Sox proteins in neuronal development and disease,along with their transcriptional changes under SUMOylation.Across three species,lysine is the conserved residue for SUMOylation.In Drosophila,SUMOylation of Sox N plays a repressive role in transcriptional activity,which impairs central nervous system development.However,de SUMOylation of Sox E and Sox11 plays neuroprotective roles,which promote neural crest precursor formation in Xenopus and retinal ganglion cell differentiation as well as axon regeneration in the rodent.We further discuss a potential translational therapy by SUMO site modification using AAV gene transduction and Clustered regularly interspaced short palindromic repeats-Cas9 technology.Understanding the underlying mechanisms of Sox SUMOylation,especially in the rodent system,may provide a therapeutic strategy to address issues associated with neuronal development and neurodegeneration.
基金supported by grants from National Key R&D Program of China(2017YFA0505801)the National Natural Science Foundation of China(81825015,81871650 and 31630086)+2 种基金National Science and Technology Major Project(2018ZX10101004)the Natural Science Foundation of Hubei Province Innovation Group(2017CFA022)Advanced Customer Cultivation Project of Wuhan National Biosafety Laboratory(2019ACCP-MS06)。
文摘Human parainfluenza virus type 3(HPIV3),a member of the Paramyxoviridae family,can cause lower respiratory disease in infants and young children.The phosphoprotein(P)of HPIV3 is an essential cofactor of the viral RNA-dependent RNA polymerase large protein(L).P connects nucleocapsid protein(N)with L to initiate genome transcription and replication.Sumoylation influences many important pathways of the target proteins,and many viral proteins are also themselves sumoylated.In this study,we found that the P of HPIV3 could be sumoylated,and mutation of K492 and K532 to arginine(PK492 R/K532 R)failed to be sumoylated within P,which enhances HPIV3 minigenome activity.Biochemical studies showed that PK492 R/K532 Rhad no effect on its interactions with N,formation of homo-tetramers and formation of inclusion bodies.Finally,we found that incorporation of K492 R/K532 R into a recombinant HPIV3(rHPIV3-PK492 R/K532 R)increased viral production in culture cells,suggesting that sumoylation attenuates functions of P and down-regulates viral replication.
基金supported by the Natural Science Foundation of Guangxi Zhuang Autonomous Region,China (No.2019JJA140728)the Horizontal Cooperation Project with Hunan Mingshun Pharmaceutical Co.,Ltd. (No.2021GLHX02)the Guangxi Province Postgraduate Co-training Base for Cooperative Innovation in Basic Medicine of Guilin Medical University (No.Gui Xue Wei[2020]7).
文摘This study aims to investigate the impact of hepatocyte nuclear factor 1β(HNF1b)on macrophage sortilin-mediated lipid metabolism and aortic atherosclerosis and explore the role of the flavone of Polygonatum odoratum(PAOA-flavone)-promoted small ubiquitin-related modifier(SUMO)modification in the atheroprotective efficacy of HNF1b.HNF1b was predicted to be a transcriptional regulator of sortilin expression via bioinformatics,dual-luciferase reporter gene assay,and chromatin immunoprecipitation.HNF1b overexpression decreased sortilin expression and cellular lipid contents in THP-1 macrophages,leading to a depression in atherosclerotic plaque formation in low-density lipoprotein(LDL)receptor-deficient(LDLR−/−)mice.Multiple SUMO1-modified sites were identified on the HNF1b protein and co-immunoprecipitation confirmed its SUMO1 modification.The SUMOylation of HNF1b protein enhanced the HNF1b-inhibited effect on sortilin expression and reduced lipid contents in macrophages.PAOA-flavone treatment promoted SUMO-activating enzyme subunit 1(SAE1)expression and SAE1-catalyzed SUMOylation of the HNF1b protein,which prevented sortilin-mediated lipid accumulation in macrophages and the formation of atherosclerotic plaques in apolipoprotein E-deficient(ApoE−/−)mice.Interference with SAE1 abrogated the improvement in lipid metabolism in macrophage cells and atheroprotective efficacy in vivo upon PAOA-flavone administration.In summary,HNF1b transcriptionally suppressed sortilin expression and macrophage lipid accumulation to inhibit aortic lipid deposition and the development of atherosclerosis.This anti-atherosclerotic effect was enhanced by PAOA-flavone-facilitated,SAE1-catalyzed SUMOylation of the HNF1b protein.
基金supported by the National Natural Science Foundation of China (81503074)
文摘Recently, Khayachi et al.;showed that fragile X mental retardation protein （FMRP） is an active substrate of the small ubiquitin-like modifier （SUMO） pathway in neurons.FMRP SUMOylation is induced by the activation of metabotropic glutamate receptors （mGlu5Rs）. FMRP
基金supported by grants from the National Natural Science Foundation of China(81770824,81270239)。
文摘Background:Abnormal myocardial voltage-gated sodium channel 1.5(Nav1.5)expression and function cause lethal ventricular arrhythmias during myocardial ischemia–reperfusion(I/R).Protein inhibitor of activated STAT Y(PIASy)-mediated caveolin-3(Cav-3)small ubiquitin-related modifier(SUMO)modification affects Cav-3 binding to the Nav1.5.PIASy activity is increased after myocardial I/R,but it is unclear whether this is attributable to plasma membrane Nav1.5 downregulation and ventricular arrhythmias.Methods:Using recombinant adeno-associated virus subtype 9(AAV9),rat cardiac PIASy was silenced using intraventricular injection of PIASy short hairpin RNA(shRNA).After two weeks,rat hearts were subjected to I/R and electrocardiography was performed to assess malignant arrhythmias.Tissues from peri-infarct areas of the left ventricle were collected for molecular biological measurements.Results:PIASy was upregulated by I/R(P<0.01),with increased SUMO2/3 modification of Cav-3 and reduced membrane Nav1.5 density(P<0.01).AAV9-PIASy shRNA intraventricular injection into the rat heart down-regulated PIASy after I/R,at both mRNA and protein levels(P<0.05 vs.Scramble-shRNA+I/R group),decreased SUMO-modified Cav-3 levels,enhanced Cav-3 binding to Nav1.5,and prevented I/R-induced decrease of Nav1.5 and Cav-3co-localization in the intercalated disc and lateral membrane.PIASy silencing in rat hearts reduced I/R-induced fatal arrhythmias,which was reflected by a modest decrease in the duration of ventricular fibrillation(VF;P<0.05 vs.Scramble-shRNA+I/R group)and a significantly reduced arrhythmia score(P<0.01 vs.Scramble-shRNA+I/R group).The anti-arrhythmic effects of PIASy silencing were also evidenced by decreased episodes of ventricular tachycardia(VT),sustained VT and VF,especially at the time 5–10 min after ischemia(P<0.05 vs.Scramble-shRNA+IR group).Using in vitro human embryonic kidney 293 T(HEK293T)cells and isolated adult rat cardiomyocyte models exposed to hypoxia/reoxygenation(H/R),we confirmed that increased PIASy promoted Cav-3 modification by SUMO2/3 and Nav1.5/Cav-3 dissociation after H/R.Mutation of SUMO consensus lysine sites in Cav-3(K38R or K144R)altered the membrane expression levels of Nav1.5 and Cav-3 before and after H/R in HEK293T cells.Conclusions:I/R-induced cardiac PIASy activation increased Cav-3 SUMOylation by SUMO2/3 and dysregulated Nav1.5-related ventricular arrhythmias.Cardiac-targeted PIASy silencing mediated Cav-3 deSUMOylation and partially prevented I/R-induced Nav1.5 downregulation in the plasma membrane of cardiomyocytes,and subsequent ventricular arrhythmias in rats.PIASy was identified as a potential therapeutic target for life-threatening arrhythmias in patients with ischemic heart diseases.
文摘Objective This study elucidated the function and role of SUMOylation in type Ⅰ endometrial carcinoma.Methods Fifty type Ⅰ endometrial carcinoma cases and para-cancer tissue samples were collected.The expression levels of ubiquitin-conjugating enzyme E2I(Ube2 i,Ubc9) and small ubiquitin-like modifier 1(SUMO1)/sentrin-specific peptidase 1(SENP1) proteins were examined using immunohistochemistry and the correlation with clinicopathological parameters was analyzed.Results Ubc9 expression in type Ⅰ endometrial carcinoma tissues was significantly higher than that in the para-cancer tissues;in contrast,the expression of the SENP1 protein was markedly lower than that in the para-cancer tissues.Ubc9 and SENP1 expression levels were negatively correlated and were associated with tumor differentiation,but not age,depth of invasion,tumor stage,and lymph node metastasis.Conclusion SUMOylation modification plays a major role in the pathogenesis and development of type Ⅰ endometrial carcinoma.Thus,it could be a potential target for the treatment of endometrial cancer.
文摘Dear Editor, In recent years, post-translational modifications （PTMs） by small ubiquitin-related modifiers （SUMOs） have emerged as an important regulatory mechanism for both cellular and viral processes （Ribet and Cossart, 2010）. Identifying the SUMOylation sites of the target protein is important to understand the molecular mechanism under- lying SUMO modification and virus-host interactions, as well as provide new insights into antiviral drug develop- ment （Wimmer and Schreiner, 2015）. Traditional site- directed mutagenesis for identifying viral protein SUMO- ylation sites lacks a specific aim and is laborious （McManus et al., 2016）. Recently, mass spectrometry （MS） has been employed as an accurate and sensitive tool to identify PTM sites, thereby greatly expanding the number of known SUMOylated proteins （Pedrioli et al., 2006）. However, during viral infection, SUMOylation is highly dynamic and SUMOylated viral proteins often have low abundance, which makes studying SUMOylation under natural conditions difficult.
文摘Recently there have been exciting research advances in neuroprotective therapies for ischemic stroke. In the past, the search for neu- roprotective agents has been fraught with failure at the clinical trials stage due to numerous factors, including subject heterogeneity and improper therapeutic windows (Tymianski, 2017). Moreover, it is becoming clearer that the complex and evolving pathobiology of stroke requires multimodal therapeutic approaches capable of modulating the numerous axes that contribute to ischemia/reperfusion damage, rather than targeting a single axis (Bernstock et al., 2018a). With the success of recent endovascular thrombectomy (EVT) trials, it has been suggested that clinical trials of EVT with adjunct neuroprotection can overcome past difficulties and maximize the effect size by using imaging to reduce patient heterogeneity (i. e., selecting those with large vessel occlusions, small ischemic cores, and good collateral circulation), restoring perfusion using better EVT devices, and enrolling patients in the correct therapeutic window (i.e., when they still have salvageable brain tissue) (Tymianski, 2017). Considering the opportunity that this represents for new, better clinical trials of neuroprotective agents, the search is on for high-potential compounds that may be investigated in these future studies.
基金supported by Deutsche Forschungsgemeinschaft(DFG)grant DFG LE 2299/5-1J.V.acknowledges support from ProteoCure COST(European Cooperation in Science and Technology)Action CA20113+1 种基金which funded the research stay during which the methodology applied in this publication was acquired.J.R.C.was supported by DFG grant DFG LE 2299/8-1.J.J.P.was supported by the Czech Science Foundation(GA23-05284S)The CEITEC Proteomics Core Facility of CIISB,Instruct-CZ Centre,was supported by MEYS CR(LM2023042,CZ.02.01.01/00/23_015/0008175,and e-INFRA CZ[ID:90254]).
文摘The centromere is a specialized domain that facilitates chromosome segregation during mitosis and serves as the site of kinetochore formation.KINETOCHORE NULL2(αKNL2)is essential for the recognition and loading of the centromeric histone H3 variant CENH3 at centromeres.A yeast two-hybrid screen forαKNL2 interactors identified components of the SUMOylation pathway.However,the role ofαKNL2 SUMOylation in Arabidopsis has not yet been determined.In this study,we demonstrated that the C-terminal region ofαKNL2(designatedαKNL2-C)interacts with small ubiquitin-like modifier 3(SUMO3)and ULP1d,as shown by bimolecular fluorescence complementation and co-immunoprecipitation assays.Bioinformatic and functional analyses ofαKNL2-C identified three SUMOylation sites and two SUMOinteracting motifs,which were shown to be critical for growth,fertility,and chromosome alignment.Of the three SUMOylation sites,Lys474 and Lys511 are the most critical for the centromeric localization ofαKNL2,underscoring the importance ofαKNL2 SUMOylation for its function.Additionally,both in vitro and in vivo assays showed thatαKNL2-C undergoes SUMOylation by SUMO1 or SUMO3.The Arabidopsis SUMO protease mutant ulp1d-2 exhibits a mild accumulation of SUMOylatedαKNL2.We further showed that SUMOylation ofαKNL2 promotes its binding to CENH3 and controls protein stability.Our findings demonstrate that C-terminal SUMOylation ofαKNL2 is crucial for its centromeric localization,interaction with CENH3,and kinetochore assembly,emphasizing the significance of post-translational modifications in chromosome segregation and cell division in plants.
基金supported by grants from the National Key Research and Development Plan of China(2021YFF1000103)the National Natural Science Foundation of China(32470339 and 32072095)+1 种基金the Ministry of Education(MOE)of Singapore Tier 2 Program(MOE-T2EP30123-0001 to W.M.)the Asian Institute of Modern Agronomy(AIMA)R&D Funding Program(AIMA-2024-WP2 to W.M.)。
文摘SUMO PROTEASE RELATED TO FERTILITY 1(SPF1)and SPF2 are responsible for deSUMOylation of SUMO-conjugated protein substrates and for maintaining protein SUMOylation homeostasis.To date,the role of SUMO proteases in fatty acid biosynthesis and lipid accumulation remains unclear.Here,we demonstrate that the Arabidopsis thaliana mutants spf1-1,spf2-1,and spf1-1 spf2-1 exhibit increased seed size and elevated seed oil content(SOC).We further show that SPF1 and SPF2 interact with WRINKLED1(WRI1),a master regulator of the transcriptional control of plant oil synthesis.Genetic analyses indicate that the spf1-1 wri1-3 and spf2-1 wri1-3 double mutants,as well as the spf1-1 spf2-1 wri1-3 triple mutant,phenocopy wri1-3 and display severe seed shriveling,indicating that SPF1 and SPF2 act upstream of WRI1.WRI1 was identified as a SUMO1 substrate with two conserved SUMOylation sites,lysine 257(K257)and K266,in cruciferous plants,with K257 acting as the dominant site required for seed oil synthesis.SUMOylation enhances WRI1 stability,whereas SPF1and SPF2mediated deSUMOylation promotes WRI1 degradation.In spf1-1,spf2-1,and spf1-1 spf2-1 mutants,the abundance of SUMOylated WRI1 increases during seed development and correlates with elevated seed oil accumulation.Together,these results indicate that SPF1 and SPF2 negatively regulate oil synthesis by deSUMOylating WRI1,establishing a dynamic SUMOylation and deSUMOylation switch centered on the SPF1/SPF2-WRI1 module that fine-tunes seed development and oil synthesis.
基金supported by National Natural Science Foundation of China(82341230 to Ye Yang)Basic Research Plan(Natural Science Foundation)in“Climbing”Program of Jiangsu Province(BK20240003 to Ye Yang,China)+1 种基金Science and Technology Development Plan Project of Jiangsu Provincial Administration of Traditional Chinese Medicine(ZD202203 to Chunyan Gu,China)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(Traditional Chinese Medicine,Integration of Chinese and Western Medicine)(China).
文摘Most cancers are currently incurable,partly due to abnormal post-translational modifications(PTMs).In this study,we initially used multiple myeloma(MM)as a working model and found that SUMOylation activating enzyme subunit 1(SAE1)promotes the malignancy of MM.Through proteome microarray analysis,SAE1 was identified as a potential target for bioactive colcemid or its derivative colchicine.Elevated levels of SAE1 were associated with poor clinical survival and increased MM proliferation in vitro and in vivo.Additionally,SAE1 directly SUMOylated and upregulated the total protein expression of p27,leading to LLPS-mediated nuclear export of p27.Our study also demonstrated the involvement of SAE1 in other types of cancer cells,and provided the first monomer crystal structure of SAE1 and its key binding model with colchicine.Colchicine also showed promising results in the Patient-Derived Tumor Xenograft(PDX)model.Furthermore,a controlled clinical trial with 56 MM patients demonstrated the clinical efficacy of colchicine.Our findings reveal a novel mechanism by which tumor cells evade p27-induced cellular growth arrest through p27 SUMOylation-mediated nuclear export.SAE1 may serve as a promising therapeutic target,and colchicine may be a potential treatment option for multiple types of cancer in clinical settings.
基金supported by the Major Program of Guangdong Basic and Applied Research(2019B030302006)the National Natural Science Foundation of China(32270292,32270752)+3 种基金the Natural Science Foundation of Guangdong(2019A1515110330,2018B030308002,2024A1515011071)Guangdong Modern Agro-industry Technology Research System(2023KJ114)the Program for Changjiang Scholars,the Scientific Research Start-up Fund for Ph D of Zhaoqing University(240013)Youth Foundation of Zhaoqing University(QN202439)。
文摘SUMOylation is a reversible posttranslational modification that plays a crucial role in various essential biological processes within eukaryotic cells.This reaction involves the transfer of small ubiquitin-like modifier(SUMO)molecules to lysine residues on target proteins,thereby regulating the stability,localization,and activity of these substrates.Despite the low amino acid sequence similarity between SUMO and ubiquitin,they share similar three-dimensional structures and are conjugated onto protein substrates through a comparable three-step catalytic reaction.
基金supported by the National Natural Science Foundation of China(No.82230108,32171238,82173466,and 82273581).
文摘Chemo-/radioresistance of malignant tumors hampers cancer control and increases patient mortality.Efficient repair of damaged DNA is critical for the maintenance of genomic integrity and fidelity of genetic information.In reverse,increased DNA repair capability in cancer cells contributes to chemo-/radioresistance of malignant tumors.DNA double-strand break(DSB)is the most serious DNA damage and is also the principal molecular basis of radiotherapy.Upon DNA damage,the Ku80 is recruited and forms a critical DNA-PK complex at the DSB sites with Ku70 and the catalytic subunit(DNA-PKcs)to initiate DNA repair.How DNA-PK is assembled and activated is not fully understood.Based on the identification of radiation-reduced Ku80 K568 crotonylation through quantitative global lysine crotonylome analysis,we reveal that Ku80 K568 is crotonylated by p300-CBP-associated factor(PCAF).Upon DNA damage,the K568cr is decrotonylated by HDAC8(Histone deacetylase 8).Decrotonylation of K568cr empties this site for the subsequent SUMOylation of Ku80 by CBX4.The conversion of Ku80 from K568 crotonylation to SUMOylation facilitates the assembly of DNA-PK complex and autophosphorylation of DNA-PKcs S2056,consequently activating the DSB repair.Moreover,mutation disrupting the post-translational modification(PTM)of Ku80 K568 site sensitizes cancer cells to radiotherapy in tumorbearing nude mice models.This study elucidates the conversion model between two different forms of PTMs in the regulation of DNA-PK complex assembly and DSB repair,highlighting this model’s potential in controlling chemo-/radioresistance of malignant tumors,as well as expands the atlas of therapeutic targets.
基金supported by the Major Program of Guangdong Basic and Applied Research(2019B030302006)the National Natural Science Foundation of China(32270292,32270752,and 32200436)+2 种基金the Natural Science Foundation of Guangdong(2024A1515011071,2021A1515011151,and 2019A1515110330)the Guangdong Modern Agro-industry Technology Research System(2023KJ114)the Program for Changjiang Scholars.
文摘Dear Editor,SUMOylation is a crucial form of post-translational modification in which small ubiquitin-like modifiers(SUMOs)are transferred to lysine residues of target proteins,thereby regulating their stability,localization,and activity.Proteomic analysis in Arabidopsis has revealed numerous proteins serving as SUMOylation substrates,which play crucial roles in development and stress responses(Elrouby and Coupland,2010).
基金supported by the National Natural Science Foundation of China(No.82273457)the Guangdong Basic and Applied Basic Research Foundation of China(No.2023A1515012762 and 2021A1515010846)+2 种基金Special Grant for Key Area Programs of Guangdong Education Department(China)(No.2021ZDZX2040)Science and Technology Special Project of Guangdong Province,China(No.210715216902829)“Dengfeng Project”for the construction of high-level hospitals in Guangdong Provinced First Affiliated Hospital of Shantou University College Supporting Funding(No.202003-10).
文摘The growing interest in post-translational protein modification,particularly in SU-MOylation,is driven by its crucial role in cell cycle regulation.SUMOylation affects various cell cycle regulators,including oncogenes,suggesting its relevance in cancer.SUMO E3 ligases are pivotal in this process,exhibiting diverse functionalities through structural domains and sub-cellular localizations.A less-explored SUMO E3 ligase,RANBP2,a component of the vertebrate nuclear pore complex,emerges as a central player in cellular cycle processes,as well as in tumorigenesis.The current studies illuminate the importance of RANBP2 and underscore the need for more extensive studies to validate its clinical applicability in neoplastic interventions.Our review elucidates the significance of RANBP2 across various types of malignancies.Addi-tionally,it delves into exploring RANBP2 as a prospective therapeutic target for cancer treat-ment,offering insights into the avenues that scholars should pursue in their subsequent research endeavors.Thus,further investigation into RANBP2’s role in solid tumorigenesis is eagerly awaited.
