Deubiquitinating enzymes （DUBs） play an important role in ubiquitin-dependent processes as negative regulators of protein ubiquitination. Ubiquitin-specific protease 26 （USP26） is a member of this family. The expr...Deubiquitinating enzymes （DUBs） play an important role in ubiquitin-dependent processes as negative regulators of protein ubiquitination. Ubiquitin-specific protease 26 （USP26） is a member of this family. The expression of Usp26 in mammalian testis and in other tissues has yet to be fully elucidated. To study the expression of Usp26 mRNA and protein in various murine tissues, reverse transcription （RT）-PCR and immunohistochemistry analyses were carried out. The RT-PCR analysis showed that the Usp26 transcript was expressed in all of the tested tissues. USP26 protein localization was examined by immunohistochemistry, and it was shown that USP26 was not detectable at 20 days postpartum, with the expression restricted to the cytoplasm of condensing spermatids （steps 9-16）, Leydig cells and nerve fibers in the brain. In addition, the USP26 protein was detected at moderate levels in myocardial ceils, the corpus of epidydimis, epithelium of the renal tubules and the seminal gland of postnatal day 35 mice. Its spatial and temporal expression pattern suggests that Usp26 may play an important role in development or function of the testis and brain. Further research into these possibilities is in progress.展开更多
Purpose:To analyze the change of water-soluble-protein(WSP),urea-soluble-protein(USP)and membrane intrinsic protein(MIP)in human senile catarct.Methods:The water-soluble-fractions(WSF)were prepared basically according...Purpose:To analyze the change of water-soluble-protein(WSP),urea-soluble-protein(USP)and membrane intrinsic protein(MIP)in human senile catarct.Methods:The water-soluble-fractions(WSF)were prepared basically according to the method of Kibbelear,et al.But in this study,5mmol/LB-mercaptoethanol was added to the buffer solution.The urea-soluble-fractions(USF)were pre-pared basically according to the method of Kibbelear,et al.Lens fiber cell mem-branes were purified basically according to the method of Russell,et al.SDS-PAGE were performed according to the procedure of Laemmili,et al.using re-solving gel13%and3%stacking gel.Results:The WSPwas fractionated intoHM^+α^-,β1-3^-andγ-crystallin compo-nents.In nuclear cataractous lenses HM^+α^-and B-crystallin increase,while r-crystallin decrease.The USP from clear lenses contains mainlyαβchains of22KD,whereas in cataractous lenses,especially in nuclear cataractous lenses,the relative amount of the 28-and23KDpolypeptide(the components of β-crys-tallin)increased markedly.Lens fiber cell MIP,clear lens and cataract lens con-tained the main polypeptide of 27KD(MIP)and23KD(MP23).Conclusion:The water-insolube protein,whether in quantity or in quality,plays an important role in cataract formation.Eye Science 1995,11:124-127.展开更多
Ecdysone receptor （EcR） and ultraspiracle （USP） form heterodimers to mediate ecdysteroid signaling during molting and metamorphosis. Various EcR/USP heterodimers have been reported. However, it is unclear what kin...Ecdysone receptor （EcR） and ultraspiracle （USP） form heterodimers to mediate ecdysteroid signaling during molting and metamorphosis. Various EcR/USP heterodimers have been reported. However, it is unclear what kind of EcR/USP combination is adopted by lepidopteran insects during the larval-pupal metamorphosis and whether the EcR/USP heterodimer varies among different tissues. To address these questions, two isoforms of each EcR and USP were cloned from the common cutworm, their messenger RNA expression patterns were examined by real-time quantitative polymerase chain reaction in different tissues during the larval-pupal metamorphosis and in the midgut in response to hormonal induction. Furthermore, their subcellular localization and protein-protein interaction were explored by transient expression and far-western blotting, respectively. All the four genes were significantly up-regulated in prepuae and/or pupae. The expression profiles of EcRB1 and USP1 were nearly identical to each other in the epidermis, fat body and midgut, and a similar situation also applied to EcRA and USP2. The three genes responded to 20-hydroxyecdysone （20E） induction except for USP2, and USP1 could be up-regulated by both 20E and juvenile hormone. The four proteins mainly localized in the nucleus and the nuclear localization was promoted by 20E. The protein-protein interaction between each EcR and USP was found in vitro. These results suggest that two types of EcR/USP heterodimer （EeRA/USP2 and EcRB 1/USP1） may exist simultaneously in the common cutworm, and the latter should play more important roles during the larval-pupal metamorphosis. In addition, the types of EcR/USP heterodimer do not vary in the tissues which undergo histolysis and regeneration during metamorphosis.展开更多
Ubiquitin-specific protease(USP7),also known as Herpesvirus-associated ubiquitin-specific protease(HAUSP),is a deubiquitinase.There has been significant recent attention on USP7 following the discovery that USP7 is a ...Ubiquitin-specific protease(USP7),also known as Herpesvirus-associated ubiquitin-specific protease(HAUSP),is a deubiquitinase.There has been significant recent attention on USP7 following the discovery that USP7 is a key regulator of the p53-MDM2 pathway.The USP7 protein is 130 kDa in size and has multiple domains which bind to a diverse set of proteins.These interactions mediate key developmental and homeostatic processes including the cell cycle,immune response,and modulation of transcription factor and epigenetic regulator activity and localization.USP7 also promotes carcinogenesis through aberrant activation of the Wnt signalling pathway and stabilization of HIF-1α.These findings have shown that USP7 may induce tumour progression and be a therapeutic target.Together with interest in developing USP7 as a target,several studies have defined new protein interactions and the regulatory networks within which USP7 functions.In this review,we focus on the protein interactions of USP7 that are most important for its cancer-associated roles.展开更多
Background:Hepatocellular carcinoma(HCC)is one of the most common malignant tumors in the world.The ubiquitin-specific peptidase 25(USP25)protein has been reported to participate in the development of several cancers....Background:Hepatocellular carcinoma(HCC)is one of the most common malignant tumors in the world.The ubiquitin-specific peptidase 25(USP25)protein has been reported to participate in the development of several cancers.However,few studies have reported its association with HCC.In this study,we aimed to investigate the function and mechanism of USP25 in the progression of HCC.Methods:We analyzed USP25 protein expression in HCC based on The Cancer Genome Atlas(TCGA)and International Cancer Genome Consortium(ICGC)database cohorts.Then,we constructed USP25-overexpressing and USP25-knockdown HepG2,MHCC97H,and L-O2 cells.We detected the biological function of USP25 by performing a series of assays,such as Cell Counting Kit-8(CCK-8),colony formation,transwell,and wound healing assays.Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR)analyses were performed to detect the interaction between USP25 and the Wnt/β-catenin signaling pathway.The relationship between USP25 and tripartite motif-containing 21(TRIM21)was assessed through mass spectrometry and co-immunoprecipitation(Co-IP)analysis.Finally,we constructed a mouse liver cancer model with the USP25 gene deletion to verify in vivo role of USP25.Results:USP25 was highly expressed in HCC tissue and HCC cell lines.Importantly,high expression of USP25 in tissues was closely related to a poor prognosis.USP25 knockdown markedly reduced the proliferation,migration,and invasion of HepG2 and MHCC97H cells,whereas USP25 overexpression led to the opposite effects.In addition,we demonstrated that USP25 interacts with TRIM21 to regulate the expression of proteins related to epithelial-mesenchymal transition(EMT;E-cadherin,N-cadherin,and Snail)and the Wnt/β-catenin pathway(β-catenin,Adenomatous polyposis coli,Axin2 and Glycogen synthase kinase 3 beta)and those of their downstream proteins(C-myc and Cyclin D1).Finally,we verified that knocking out USP25 inhibited tumor growth and distant metastasis in vivo.Conclusions:In summary,our data showed that USP25 was overexpressed in HCC.USP25 promoted the proliferation,migration,invasion,and EMT of HCC cells by interacting with TRIM21 to activate theβ-catenin signaling pathway.展开更多
基金Acknowledgment We thank the laboratory, clinical and paramedical staff of the center of Reproductive Medicine, and the Department of Pathology for their assistance. This study was supported by the National Natural Science Foundation of China （30471735 and 30700654） and the Sci-Technical Development Project of Shanxi Province, China （2006K 15-G4）.
文摘Deubiquitinating enzymes （DUBs） play an important role in ubiquitin-dependent processes as negative regulators of protein ubiquitination. Ubiquitin-specific protease 26 （USP26） is a member of this family. The expression of Usp26 in mammalian testis and in other tissues has yet to be fully elucidated. To study the expression of Usp26 mRNA and protein in various murine tissues, reverse transcription （RT）-PCR and immunohistochemistry analyses were carried out. The RT-PCR analysis showed that the Usp26 transcript was expressed in all of the tested tissues. USP26 protein localization was examined by immunohistochemistry, and it was shown that USP26 was not detectable at 20 days postpartum, with the expression restricted to the cytoplasm of condensing spermatids （steps 9-16）, Leydig cells and nerve fibers in the brain. In addition, the USP26 protein was detected at moderate levels in myocardial ceils, the corpus of epidydimis, epithelium of the renal tubules and the seminal gland of postnatal day 35 mice. Its spatial and temporal expression pattern suggests that Usp26 may play an important role in development or function of the testis and brain. Further research into these possibilities is in progress.
文摘Purpose:To analyze the change of water-soluble-protein(WSP),urea-soluble-protein(USP)and membrane intrinsic protein(MIP)in human senile catarct.Methods:The water-soluble-fractions(WSF)were prepared basically according to the method of Kibbelear,et al.But in this study,5mmol/LB-mercaptoethanol was added to the buffer solution.The urea-soluble-fractions(USF)were pre-pared basically according to the method of Kibbelear,et al.Lens fiber cell mem-branes were purified basically according to the method of Russell,et al.SDS-PAGE were performed according to the procedure of Laemmili,et al.using re-solving gel13%and3%stacking gel.Results:The WSPwas fractionated intoHM^+α^-,β1-3^-andγ-crystallin compo-nents.In nuclear cataractous lenses HM^+α^-and B-crystallin increase,while r-crystallin decrease.The USP from clear lenses contains mainlyαβchains of22KD,whereas in cataractous lenses,especially in nuclear cataractous lenses,the relative amount of the 28-and23KDpolypeptide(the components of β-crys-tallin)increased markedly.Lens fiber cell MIP,clear lens and cataract lens con-tained the main polypeptide of 27KD(MIP)and23KD(MP23).Conclusion:The water-insolube protein,whether in quantity or in quality,plays an important role in cataract formation.Eye Science 1995,11:124-127.
基金Acknowledgments The research was supported by the grants from National Natural Science Foundation of China （Grant No. 31172154） and the National Basic Research Program of China （973 Program, No. 2012CB114101）.
文摘Ecdysone receptor （EcR） and ultraspiracle （USP） form heterodimers to mediate ecdysteroid signaling during molting and metamorphosis. Various EcR/USP heterodimers have been reported. However, it is unclear what kind of EcR/USP combination is adopted by lepidopteran insects during the larval-pupal metamorphosis and whether the EcR/USP heterodimer varies among different tissues. To address these questions, two isoforms of each EcR and USP were cloned from the common cutworm, their messenger RNA expression patterns were examined by real-time quantitative polymerase chain reaction in different tissues during the larval-pupal metamorphosis and in the midgut in response to hormonal induction. Furthermore, their subcellular localization and protein-protein interaction were explored by transient expression and far-western blotting, respectively. All the four genes were significantly up-regulated in prepuae and/or pupae. The expression profiles of EcRB1 and USP1 were nearly identical to each other in the epidermis, fat body and midgut, and a similar situation also applied to EcRA and USP2. The three genes responded to 20-hydroxyecdysone （20E） induction except for USP2, and USP1 could be up-regulated by both 20E and juvenile hormone. The four proteins mainly localized in the nucleus and the nuclear localization was promoted by 20E. The protein-protein interaction between each EcR and USP was found in vitro. These results suggest that two types of EcR/USP heterodimer （EeRA/USP2 and EcRB 1/USP1） may exist simultaneously in the common cutworm, and the latter should play more important roles during the larval-pupal metamorphosis. In addition, the types of EcR/USP heterodimer do not vary in the tissues which undergo histolysis and regeneration during metamorphosis.
基金This work was supported by the Medical Research Council,UK(No.MR/S01411X/1)the Saudi Arabia Cultural Bureau in London(No.DMU500).
文摘Ubiquitin-specific protease(USP7),also known as Herpesvirus-associated ubiquitin-specific protease(HAUSP),is a deubiquitinase.There has been significant recent attention on USP7 following the discovery that USP7 is a key regulator of the p53-MDM2 pathway.The USP7 protein is 130 kDa in size and has multiple domains which bind to a diverse set of proteins.These interactions mediate key developmental and homeostatic processes including the cell cycle,immune response,and modulation of transcription factor and epigenetic regulator activity and localization.USP7 also promotes carcinogenesis through aberrant activation of the Wnt signalling pathway and stabilization of HIF-1α.These findings have shown that USP7 may induce tumour progression and be a therapeutic target.Together with interest in developing USP7 as a target,several studies have defined new protein interactions and the regulatory networks within which USP7 functions.In this review,we focus on the protein interactions of USP7 that are most important for its cancer-associated roles.
基金National Natural Science Foundation of China(No.81870392)
文摘Background:Hepatocellular carcinoma(HCC)is one of the most common malignant tumors in the world.The ubiquitin-specific peptidase 25(USP25)protein has been reported to participate in the development of several cancers.However,few studies have reported its association with HCC.In this study,we aimed to investigate the function and mechanism of USP25 in the progression of HCC.Methods:We analyzed USP25 protein expression in HCC based on The Cancer Genome Atlas(TCGA)and International Cancer Genome Consortium(ICGC)database cohorts.Then,we constructed USP25-overexpressing and USP25-knockdown HepG2,MHCC97H,and L-O2 cells.We detected the biological function of USP25 by performing a series of assays,such as Cell Counting Kit-8(CCK-8),colony formation,transwell,and wound healing assays.Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR)analyses were performed to detect the interaction between USP25 and the Wnt/β-catenin signaling pathway.The relationship between USP25 and tripartite motif-containing 21(TRIM21)was assessed through mass spectrometry and co-immunoprecipitation(Co-IP)analysis.Finally,we constructed a mouse liver cancer model with the USP25 gene deletion to verify in vivo role of USP25.Results:USP25 was highly expressed in HCC tissue and HCC cell lines.Importantly,high expression of USP25 in tissues was closely related to a poor prognosis.USP25 knockdown markedly reduced the proliferation,migration,and invasion of HepG2 and MHCC97H cells,whereas USP25 overexpression led to the opposite effects.In addition,we demonstrated that USP25 interacts with TRIM21 to regulate the expression of proteins related to epithelial-mesenchymal transition(EMT;E-cadherin,N-cadherin,and Snail)and the Wnt/β-catenin pathway(β-catenin,Adenomatous polyposis coli,Axin2 and Glycogen synthase kinase 3 beta)and those of their downstream proteins(C-myc and Cyclin D1).Finally,we verified that knocking out USP25 inhibited tumor growth and distant metastasis in vivo.Conclusions:In summary,our data showed that USP25 was overexpressed in HCC.USP25 promoted the proliferation,migration,invasion,and EMT of HCC cells by interacting with TRIM21 to activate theβ-catenin signaling pathway.
