Neddylation is a crucial posttranslational modification that involves the attachment of neural precursor cell-expressed developmentally downregulated protein 8(NEDD8)to a lysine residue in the substrate via the sequen...Neddylation is a crucial posttranslational modification that involves the attachment of neural precursor cell-expressed developmentally downregulated protein 8(NEDD8)to a lysine residue in the substrate via the sequential actions of the E1 NEDD8-activating enzyme(NAE)(E1),E2 NEDD8-conjugating enzyme(E2),and E3 NEDD8-ligase(E3).The most extensively studied substrates of neddylation are members of the cullin family,which act as scaffold components for cullin ring E3 ubiquitin ligases(CRLs).Since cullin neddylation activates CRLs,which are frequently overactive in tumors,inhibiting neddylation has emerged as a promising strategy for developing novel antitumor therapies.This review explores the antitumor effects of inhibiting neddylation that leads to the inactivation of CRLs and provides a summary of known inhibitors that target protein-protein interactions(PPIs)within the neddylation enzymatic cascade.展开更多
Much akin to ubiquitylation,neddylation is catalyzed by a cascade of three enzymes:E1 NEDD8-activating enzyme,E2 NEDD8-conjugating enzyme(UBE2M or UBE2F),and E3 NEDD8 ligases.The best-known neddylation substrates are ...Much akin to ubiquitylation,neddylation is catalyzed by a cascade of three enzymes:E1 NEDD8-activating enzyme,E2 NEDD8-conjugating enzyme(UBE2M or UBE2F),and E3 NEDD8 ligases.The best-known neddylation substrates are the members of cullin family,leading to the activation of Cullin-RING ligases,which regulate a variety of downstream biological processes largely via promoting ubiquitylation and subsequent proteasomal degradation of many key signaling proteins.Notably,neddylation enzymes and components of the Cullin-RING ligases are frequently altered in many human cancers and have been validated as promising cancer targets.As such,drug discovery efforts are underway to target neddylation-Cullin-RING ligases with a few selective small molecule inhibitors being advanced into various phases of clinical trials.This review firstly provides a brief introduction to neddylation,then focuses on lung cancer,and summarizes a wealth of current data showing how neddylation-Cullin-RING ligases are altered and affect the growth and survival of lung cancer cells,lung tumorigenesis,lung tumor microenvironment,and inflammatory response.A few reported small molecule inhibitors of neddylation enzymes as well as their activity against lung cancer cells are also summarized,and future perspectives with an ultimate goal of discovering effective treatment of lung cancer via targeting neddylation-Cullin-RING ligases are proposed.展开更多
Hepatocellular carcinoma(HCC)is the most prevalent type of malignant liver tu-mor with high morbidity and mortality and severely threatens human health and life quality.Thus,it is of great significance to investigate ...Hepatocellular carcinoma(HCC)is the most prevalent type of malignant liver tu-mor with high morbidity and mortality and severely threatens human health and life quality.Thus,it is of great significance to investigate the molecular mechanism underlying the patho-genesis of HCC and seek biomarkers for early diagnosis.Neddylation,one of the most conserved post-translational modification types in eukaryotes,plays vital roles in the progres-sion of HCC.During the process of neddylation,NEDD8 is covalently conjugated to its substrate proteins,thereby modulating multiple necessary biological processes.Currently,increasing ev-idence shows that the aberrant activation of neddylation is positively correlated with the occurrence and development of tumors and the poor clinical prognosis of HCC patients.Based on the current investigations,neddylation modification has been reported to target both the cullins and non-cullin substrates and subsequently affect HCC progression,including the virus infection,malignant transformation,tumor cell proliferation,migration and invasion ability,and tumor microenvironment.Therefore,inhibitors targeting the neddylation cascade have been developed and entered clinical trials,indicating satisfactory anti-HCC treatment effects.This review aims to summarize the latest progress in the molecular mechanism of pathologi-cally aberrant neddylation in HCC,as well as the advances of neddylation-targeted inhibitors as potential drugs for HCC treatment.展开更多
Protein neddylation is a post-translational modification which transfers the ubiquitin-like protein NEDD8 to a lysine residue of the target substrate through a three-step enzymatic cascade.The bestknown substrates of ...Protein neddylation is a post-translational modification which transfers the ubiquitin-like protein NEDD8 to a lysine residue of the target substrate through a three-step enzymatic cascade.The bestknown substrates of neddylation are cullin family proteins,which are the core component of Cullin-RING E3 ubiquitin ligases(CRLs).Given that cullin neddylation is required for CRL activity,and CRLs control the turn-over of a variety of key signal proteins and are often abnormally activated in cancers,targeting neddylation becomes a promising approach for discovery of novel anti-cancer therapeutics.In the past decade,we have witnessed significant progress in the field of protein neddylation from preclinical target validation,to drug screening,then to the clinical trials of neddylation inhibitors.In this review,we first briefly introduced the nature of protein neddylation and the regulation of neddylation cascade,followed by a summary of all reported chemical inhibitors of neddylation enzymes.We then discussed the structure-based targeting of protein-protein interaction in neddylation cascade,and finally the available approaches for the discovery of new neddylation inhibitors.This review will provide a focused,up-to-date and yet comprehensive overview on the discovery effort of neddylation inhibitors.展开更多
Protein neddylation is catalyzed by a three-enzyme cascade,namely an E1 NEDD8-activating enzyme(NAE),one of two E2 NEDD8 conjugation enzymes and one of several E3 NEDD8 ligases.The physiological substrates of neddylat...Protein neddylation is catalyzed by a three-enzyme cascade,namely an E1 NEDD8-activating enzyme(NAE),one of two E2 NEDD8 conjugation enzymes and one of several E3 NEDD8 ligases.The physiological substrates of neddylation are the family members of cullin,the scaffold component of cullin RING ligases(CRLs).Currently,a potent E1 inhibitor,MLN4924,also known as pevonedistat,is in several clinical trials for anti-cancer therapy.Here we report the discovery,through virtual screening and structural modifications,of a small molecule compound HA-1141 that directly binds to NAE in both in vitro and in vivo assays and effectively inhibits neddylation of cullins 1 e5.Surprisingly,unlike MLN4924,HA-1141 also triggers non-canonical endoplasmic reticulum(ER)stress and PKR-mediated terminal integrated stress response(ISR)to activate ATF4 at an early stage,and to inhibit protein synthesis and mTORC1 activity at a later stage,eventually leading to autophagy induction.Biologically,HA-1141 suppresses growth and survival of cultured lung cancer cells and tumor growth in in vivo xenograft lung cancer models at a well-tolerated dose.Taken together,our study has identified a small molecule compound with the dual activities of blocking neddylation and triggering ER stress,leading to growth suppression of cancer cells.展开更多
NEDDylation has been shown to participate in the DNA damage pathway, but the substrates of neural precursor cell expressed developmentally downregulated 8 （NEDD8） and the roles of NEDDylation involved in the DNA dam...NEDDylation has been shown to participate in the DNA damage pathway, but the substrates of neural precursor cell expressed developmentally downregulated 8 （NEDD8） and the roles of NEDDylation involved in the DNA damage response （DDR） are largely unknown. Translesion synthesis （TLS） is a damage-tolerance mechanism, in which RAD181RAD6-mediated monoubiq- uitinated proliferating cell nuclear antigen （PCNA） pro- motes recruitment of polymerase q （polq） to bypass lesions. Here we identify PCNA as a substrate of NEDD8, and show that E3 ligase RAD18-catalyzed PCNA NEDDylation antagonizes its ubiquitination. In addition, NEDP1 acts as the deNEDDylase of PCNA, and NEDP1 deletion enhances PCNA NEDDylation but reduces its ubiquitination. In response to H202 stimulation, NEDP1 disassociates from PCNA and RAD18-dependent PCNA NEDDylation increases markedly after its ubiquitination. impairment of NEDDylation by Ubc12 knockout enhances PCNA ubiquitination and promotes PCNA-polη interaction, while up-regulation of NEDDylation by NEDD8 overexpression or NEDP1 deletion reduces the excessive accumulation of ubiquitinated PCNA, thus inhibits PCNA-polη interaction and blocks polη foci formation. Moreover, Ubc12 knockout decreases cell sensitivity to H2O2-induced oxidative stress, but NEDP1 deletion aggravates this sensitivity. Collectively, our study elucidates the important role of NEDDylation in the DDR as a modulator of PCNA monoubiquitination and polη recruitment.展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.:82060675 and 82260036)the Guizhou Provincial Science and Technology Projects,China(Grant Nos.:QKHCG[2024]ZD012 and QKHJCZK[2024]YB306)+1 种基金the Guizhou Provincial High-Level(“Thousand”Level)Innovative Talents Projects,China(Grants Nos:gzwjrs2023-034 and gzwjrs2023-041)the Zunyi Science and Technology Plan Project,China(Grant Nos.:ZSKRPT-2023-6 and ZSKHZ-2023-219).
文摘Neddylation is a crucial posttranslational modification that involves the attachment of neural precursor cell-expressed developmentally downregulated protein 8(NEDD8)to a lysine residue in the substrate via the sequential actions of the E1 NEDD8-activating enzyme(NAE)(E1),E2 NEDD8-conjugating enzyme(E2),and E3 NEDD8-ligase(E3).The most extensively studied substrates of neddylation are members of the cullin family,which act as scaffold components for cullin ring E3 ubiquitin ligases(CRLs).Since cullin neddylation activates CRLs,which are frequently overactive in tumors,inhibiting neddylation has emerged as a promising strategy for developing novel antitumor therapies.This review explores the antitumor effects of inhibiting neddylation that leads to the inactivation of CRLs and provides a summary of known inhibitors that target protein-protein interactions(PPIs)within the neddylation enzymatic cascade.
基金supported by the funds from National Natural Science Foundation of China(92253203 and U22A20317)to Y.S.Zhejiang Provincial Natural Science Foundation of China(LD22H300003)to Y.S.+1 种基金Leading Innovative and Entrepreneur Team Introduction Program of Zhejiang(2022R01002)to Y.S.a grant from Research Center for Life Science and Human Health,Binjiang Institute of Zhejiang University(ZY202205SMKY007)to Y.S.
文摘Much akin to ubiquitylation,neddylation is catalyzed by a cascade of three enzymes:E1 NEDD8-activating enzyme,E2 NEDD8-conjugating enzyme(UBE2M or UBE2F),and E3 NEDD8 ligases.The best-known neddylation substrates are the members of cullin family,leading to the activation of Cullin-RING ligases,which regulate a variety of downstream biological processes largely via promoting ubiquitylation and subsequent proteasomal degradation of many key signaling proteins.Notably,neddylation enzymes and components of the Cullin-RING ligases are frequently altered in many human cancers and have been validated as promising cancer targets.As such,drug discovery efforts are underway to target neddylation-Cullin-RING ligases with a few selective small molecule inhibitors being advanced into various phases of clinical trials.This review firstly provides a brief introduction to neddylation,then focuses on lung cancer,and summarizes a wealth of current data showing how neddylation-Cullin-RING ligases are altered and affect the growth and survival of lung cancer cells,lung tumorigenesis,lung tumor microenvironment,and inflammatory response.A few reported small molecule inhibitors of neddylation enzymes as well as their activity against lung cancer cells are also summarized,and future perspectives with an ultimate goal of discovering effective treatment of lung cancer via targeting neddylation-Cullin-RING ligases are proposed.
基金supported by the Natural Science Foundation of Shandong Province,China(No.ZR2021QC030,ZR2022LZL006)the Innovation Project of Shandong First Medical University.
文摘Hepatocellular carcinoma(HCC)is the most prevalent type of malignant liver tu-mor with high morbidity and mortality and severely threatens human health and life quality.Thus,it is of great significance to investigate the molecular mechanism underlying the patho-genesis of HCC and seek biomarkers for early diagnosis.Neddylation,one of the most conserved post-translational modification types in eukaryotes,plays vital roles in the progres-sion of HCC.During the process of neddylation,NEDD8 is covalently conjugated to its substrate proteins,thereby modulating multiple necessary biological processes.Currently,increasing ev-idence shows that the aberrant activation of neddylation is positively correlated with the occurrence and development of tumors and the poor clinical prognosis of HCC patients.Based on the current investigations,neddylation modification has been reported to target both the cullins and non-cullin substrates and subsequently affect HCC progression,including the virus infection,malignant transformation,tumor cell proliferation,migration and invasion ability,and tumor microenvironment.Therefore,inhibitors targeting the neddylation cascade have been developed and entered clinical trials,indicating satisfactory anti-HCC treatment effects.This review aims to summarize the latest progress in the molecular mechanism of pathologi-cally aberrant neddylation in HCC,as well as the advances of neddylation-targeted inhibitors as potential drugs for HCC treatment.
基金the financial support by the National Key R&D Program of China(2016YFA0501800 to YS)
文摘Protein neddylation is a post-translational modification which transfers the ubiquitin-like protein NEDD8 to a lysine residue of the target substrate through a three-step enzymatic cascade.The bestknown substrates of neddylation are cullin family proteins,which are the core component of Cullin-RING E3 ubiquitin ligases(CRLs).Given that cullin neddylation is required for CRL activity,and CRLs control the turn-over of a variety of key signal proteins and are often abnormally activated in cancers,targeting neddylation becomes a promising approach for discovery of novel anti-cancer therapeutics.In the past decade,we have witnessed significant progress in the field of protein neddylation from preclinical target validation,to drug screening,then to the clinical trials of neddylation inhibitors.In this review,we first briefly introduced the nature of protein neddylation and the regulation of neddylation cascade,followed by a summary of all reported chemical inhibitors of neddylation enzymes.We then discussed the structure-based targeting of protein-protein interaction in neddylation cascade,and finally the available approaches for the discovery of new neddylation inhibitors.This review will provide a focused,up-to-date and yet comprehensive overview on the discovery effort of neddylation inhibitors.
基金National Key R&D Program of China(2016YFA0501800 to Yi Sun)for financial support。
文摘Protein neddylation is catalyzed by a three-enzyme cascade,namely an E1 NEDD8-activating enzyme(NAE),one of two E2 NEDD8 conjugation enzymes and one of several E3 NEDD8 ligases.The physiological substrates of neddylation are the family members of cullin,the scaffold component of cullin RING ligases(CRLs).Currently,a potent E1 inhibitor,MLN4924,also known as pevonedistat,is in several clinical trials for anti-cancer therapy.Here we report the discovery,through virtual screening and structural modifications,of a small molecule compound HA-1141 that directly binds to NAE in both in vitro and in vivo assays and effectively inhibits neddylation of cullins 1 e5.Surprisingly,unlike MLN4924,HA-1141 also triggers non-canonical endoplasmic reticulum(ER)stress and PKR-mediated terminal integrated stress response(ISR)to activate ATF4 at an early stage,and to inhibit protein synthesis and mTORC1 activity at a later stage,eventually leading to autophagy induction.Biologically,HA-1141 suppresses growth and survival of cultured lung cancer cells and tumor growth in in vivo xenograft lung cancer models at a well-tolerated dose.Taken together,our study has identified a small molecule compound with the dual activities of blocking neddylation and triggering ER stress,leading to growth suppression of cancer cells.
基金We sincerely thank Profs. Jun Huang, Wensheng Wei, and Caixia Guo for providing the plasmids used in this study. We thank Millennium Pharmaceuticals for providing the MLN4924 used in this study. This work was supported by the National Natural Science Foundation of China （Grant Nos. 31470754, 81730080 and 31670786）, the National Key Research and Development Program of China （2016YFC1302401）.
文摘NEDDylation has been shown to participate in the DNA damage pathway, but the substrates of neural precursor cell expressed developmentally downregulated 8 （NEDD8） and the roles of NEDDylation involved in the DNA damage response （DDR） are largely unknown. Translesion synthesis （TLS） is a damage-tolerance mechanism, in which RAD181RAD6-mediated monoubiq- uitinated proliferating cell nuclear antigen （PCNA） pro- motes recruitment of polymerase q （polq） to bypass lesions. Here we identify PCNA as a substrate of NEDD8, and show that E3 ligase RAD18-catalyzed PCNA NEDDylation antagonizes its ubiquitination. In addition, NEDP1 acts as the deNEDDylase of PCNA, and NEDP1 deletion enhances PCNA NEDDylation but reduces its ubiquitination. In response to H202 stimulation, NEDP1 disassociates from PCNA and RAD18-dependent PCNA NEDDylation increases markedly after its ubiquitination. impairment of NEDDylation by Ubc12 knockout enhances PCNA ubiquitination and promotes PCNA-polη interaction, while up-regulation of NEDDylation by NEDD8 overexpression or NEDP1 deletion reduces the excessive accumulation of ubiquitinated PCNA, thus inhibits PCNA-polη interaction and blocks polη foci formation. Moreover, Ubc12 knockout decreases cell sensitivity to H2O2-induced oxidative stress, but NEDP1 deletion aggravates this sensitivity. Collectively, our study elucidates the important role of NEDDylation in the DDR as a modulator of PCNA monoubiquitination and polη recruitment.
