Objective: To examine the effect of pSer9-GSK-3β on breast cancer and to determine whether the underlying metabolic and immunological mechanism is associated with ROS/eIF2B and natural killer(NK) cells.Methods: We em...Objective: To examine the effect of pSer9-GSK-3β on breast cancer and to determine whether the underlying metabolic and immunological mechanism is associated with ROS/eIF2B and natural killer(NK) cells.Methods: We employed TWS119 to inactivate GSK-3β by phosphorylating Ser9 and explored its effect on breast cancer and NK cells. The expression of GSK-3β, natural killer group 2 member D(NKG2D) ligands, eIF2B was quantified by PCR and Western blot. We measured intracellular reactive oxygen species(ROS) and mitochondrial ROS using DCFH-DA and MitoSOX^(TM) probe,respectively, and conducted quantitative analysis of cellular respiration on 4T1 cells with mitochondrial respiratory chain complex Ⅰ/Ⅲ kits.Results: Our investigation revealed that TWS119 downregulated NKG2D ligands(H60 a and Rae1), suppressed the cytotoxicity of NK cells, and promoted the migration of 4T1 murine breast cancer cells. Nevertheless, LY290042, which attenuates p-GSK-3β formation by inhibiting the PI3K/Akt pathway, reversed these effects. We also found that higher expression of p Ser9-GSK-3β induced higher levels of ROS, and observed that abnormality of mitochondrial respiratory chain complex Ⅰ/Ⅲ function induced the dysfunction of GSK-3β-induced electron transport chain, naturally disturbing the ROS level. In addition, the expression of NOX3 and NOX4 was significantly up-regulated, which affected the generation of ROS and associated with the metastasis of breast cancer. Furthermore, we found that the expression of pSer535-eIF2B promoted the expression of NKG2D ligands(Mult-1 and Rae1) following by expression of pSer9-GSK-3β and generation of ROS.Conclusions: The PI3K/Akt/GSK-3β/ROS/eIF2B pathway could regulate NK cell activity and sensitivity of tumor cells to NK cells,which resulted in breast cancer growth and lung metastasis. Thus, GSK-3β is a promising target of anti-tumor therapy.展开更多
该文报道2例白质消融性白质脑病(leukoencephalopathy with vanishing white matter,VWM)。患者1,女,2岁3个月,临床以反复感染后运动能力倒退为主要表现,病情进展迅速,发病6个月时有癫痫发作,此后患者不能独立行走,吞咽困难;基因检测发...该文报道2例白质消融性白质脑病(leukoencephalopathy with vanishing white matter,VWM)。患者1,女,2岁3个月,临床以反复感染后运动能力倒退为主要表现,病情进展迅速,发病6个月时有癫痫发作,此后患者不能独立行走,吞咽困难;基因检测发现,该患者的EIF2B4基因存在7号外显子c.594C>G (p.I98M)和11号外显子c.1177T>A(p.Y393N)的复合杂合突变,其中前者来自父亲、后者来自母亲,且2个位点均是未曾报道的新错义突变。患者2,女,41岁,临床以进行性双下肢无力及记忆力减退为主要表现;基因检测发现,该患者的EIF2B3基因存在2号外显子c.130G>A(p.G44K)和8号外显子c.934C>G (p.R312G)的复合杂合突变,其中后者位点的突变已有报道,但此突变类型为首次报告,且该患者是中国报道的第2例成人型VWM。2例患者的头颅磁共振成像均表现为弥漫对称性脑白质病变;其中,患者1白质稀薄更突出,患者2主要表现为白质萎缩、脑室扩大。发病年龄是VWM严重程度的临床预测因子,起病越早则病情进展越迅速。应激是发病及神经恶化的诱因,目前VWM尚无有效的治疗方法。该文对上述2个病例进行报道,旨在提升临床医师对疾病的认识及早期诊断的能力,以期延缓疾病的进展、延长患者的生存期。展开更多
Mutations in the eIF2B gene cause the VWM disease. Genetic and biochemical data of MS patient and MRI data showing VWM images similar to MS lesions, encouraged the present study in which we analyzed the eIF2B5 gene in...Mutations in the eIF2B gene cause the VWM disease. Genetic and biochemical data of MS patient and MRI data showing VWM images similar to MS lesions, encouraged the present study in which we analyzed the eIF2B5 gene in 225 unrelated MS patients to evaluate an overlapping between MS and VWM. A common variation Ile587Val was found very frequent in the MS patients respect normal controls, thus suggesting that Ile587Val should be considered as susceptibility factor in the development of MS. In conclusion, our data strongly highlight a possible involvement of the eIF2B5 in the development of MS.展开更多
Eukaryotic translation initiation factor eIF2B, the guanine nucleotide exchange factor (GEF) for eIF2, catalyzes conversion of eIF2·GDP to eIF2·GTP. Since translation initiation can
In translation initiation, eIF2 is responsible for delivering initiator tRNA to ribosome, and such delivery only occurs when GTP is bound to eIF2. eIF2B, a guanine nucleotide
Eukaryotic translation initiation factor eIF2B,the guanine nucleotide exchange factor(GEF)for eIF2,catalyzes conversion of eIF2·GDP to eIF2·GTP.The eIF2B is composed of five subunits,α,β,γ,δandε,within ...Eukaryotic translation initiation factor eIF2B,the guanine nucleotide exchange factor(GEF)for eIF2,catalyzes conversion of eIF2·GDP to eIF2·GTP.The eIF2B is composed of five subunits,α,β,γ,δandε,within which theεsubunit is responsible for catalyzing the guanine exchange reaction.Here we present the crystal structure of the C-terminal domain of human eIF2Bε(eIF2Bε-CTD)at 2.0-Åresolution.The structure resembles a HEAT motif and three charge-rich areas on its surface can be identified.When compared to yeast eIF2Bε-CTD,one area involves highly conserved AA boxes while the other two are only partially conserved.In addition,the previously reported mutations in human eIF2Bε-CTD,which are related to the loss of the GEF activity and human VWM disease,have been discussed.Based on the structure,most of such mutations tend to destabilize the HEAT motif.展开更多
自质消融性自质脑病（leukoencephalopathy with vanishing white matter，VWM）又称儿童共济失调伴中枢神经系统髓鞘化不良（childhood ataxia with central nervous system hypomyelination，CACH，OMIM306896），是一种常染色体隐性...自质消融性自质脑病（leukoencephalopathy with vanishing white matter，VWM）又称儿童共济失调伴中枢神经系统髓鞘化不良（childhood ataxia with central nervous system hypomyelination，CACH，OMIM306896），是一种常染色体隐性遗传的向质脑病，是儿童遗传性自质脑病中常见的类型之一，其患病率尚缺乏统计。展开更多
Vanishing white matter disease （VWM）, a human atitosomal recessive inherited leukoencephalopathy, is due to mutations in eukaryotic initiation factor 2B （elF2B）. elF2B is responsible for tile initiation of protein...Vanishing white matter disease （VWM）, a human atitosomal recessive inherited leukoencephalopathy, is due to mutations in eukaryotic initiation factor 2B （elF2B）. elF2B is responsible for tile initiation of protein synthesis by its guanine nucleotide exchange lhctor （GEF） activity. Mutations ofelF2B impair GEF activity at different degree. Previous studies implied improperly activated unlblded protein response （UPR） and endoplasmic reticulum stress （ERS） participated in the pathogenesis ofVWM. Autophagy relieves endoplasmic reticulum load by eliminating the unfolded protein. It is still unknown the effects of genotypes on the pathogenesis. In this work, UPR and autophagy flux were analyzed with different mutational types. Methods： ERS tolerance, reflected by apoptosis and cell viability, was detected in human oligodendrocyte cell line transfected with the wild type, or different mutations of p. Argl 13 His, p. Arg269＊ or p. Ser610-Asp613del in el F2 Be. A representative U PR-PERK component of activating transcription lhctor 4 （ATF4） was measured under the basal condition and ERS induction. Autophagy was analyzed the flux in the presence of lysosomal inhibitors. Results： The degree of ERS tolerance varied in different genotypes. The truncated or deletion mutant showed prominent apoptosis cell viability declination after ERS induction. The most seriously damaged GEF activity ofp. Arg269＊ group underwent spontaneous apoptosis. The truncated or deletion mutant showed elevated ATF4 under basal as well as ERS condition. Decreased expression of LC3-1 and LC3-11 in the mutants reflected an impaired autophagy flux, which was more obvious in the truncated or deletion mutants alter ERS induction. Conclusions： GEF activities in dilt;erent genotypes could influence the cell ERS tolerance as well as compensatory pathways of UPR and autophagy. Oligodendrocytes with truncated or deletion inutants showed less tolerable to ERS.展开更多
基金supported by grants from the National Natural Science Foundation of China (Grant No. 8117975 and 31770968)Tianjin Institutes for Basic Sciences (Grant No. 15JCYBJC26900 and 16JCQNJC11700)
文摘Objective: To examine the effect of pSer9-GSK-3β on breast cancer and to determine whether the underlying metabolic and immunological mechanism is associated with ROS/eIF2B and natural killer(NK) cells.Methods: We employed TWS119 to inactivate GSK-3β by phosphorylating Ser9 and explored its effect on breast cancer and NK cells. The expression of GSK-3β, natural killer group 2 member D(NKG2D) ligands, eIF2B was quantified by PCR and Western blot. We measured intracellular reactive oxygen species(ROS) and mitochondrial ROS using DCFH-DA and MitoSOX^(TM) probe,respectively, and conducted quantitative analysis of cellular respiration on 4T1 cells with mitochondrial respiratory chain complex Ⅰ/Ⅲ kits.Results: Our investigation revealed that TWS119 downregulated NKG2D ligands(H60 a and Rae1), suppressed the cytotoxicity of NK cells, and promoted the migration of 4T1 murine breast cancer cells. Nevertheless, LY290042, which attenuates p-GSK-3β formation by inhibiting the PI3K/Akt pathway, reversed these effects. We also found that higher expression of p Ser9-GSK-3β induced higher levels of ROS, and observed that abnormality of mitochondrial respiratory chain complex Ⅰ/Ⅲ function induced the dysfunction of GSK-3β-induced electron transport chain, naturally disturbing the ROS level. In addition, the expression of NOX3 and NOX4 was significantly up-regulated, which affected the generation of ROS and associated with the metastasis of breast cancer. Furthermore, we found that the expression of pSer535-eIF2B promoted the expression of NKG2D ligands(Mult-1 and Rae1) following by expression of pSer9-GSK-3β and generation of ROS.Conclusions: The PI3K/Akt/GSK-3β/ROS/eIF2B pathway could regulate NK cell activity and sensitivity of tumor cells to NK cells,which resulted in breast cancer growth and lung metastasis. Thus, GSK-3β is a promising target of anti-tumor therapy.
文摘该文报道2例白质消融性白质脑病(leukoencephalopathy with vanishing white matter,VWM)。患者1,女,2岁3个月,临床以反复感染后运动能力倒退为主要表现,病情进展迅速,发病6个月时有癫痫发作,此后患者不能独立行走,吞咽困难;基因检测发现,该患者的EIF2B4基因存在7号外显子c.594C>G (p.I98M)和11号外显子c.1177T>A(p.Y393N)的复合杂合突变,其中前者来自父亲、后者来自母亲,且2个位点均是未曾报道的新错义突变。患者2,女,41岁,临床以进行性双下肢无力及记忆力减退为主要表现;基因检测发现,该患者的EIF2B3基因存在2号外显子c.130G>A(p.G44K)和8号外显子c.934C>G (p.R312G)的复合杂合突变,其中后者位点的突变已有报道,但此突变类型为首次报告,且该患者是中国报道的第2例成人型VWM。2例患者的头颅磁共振成像均表现为弥漫对称性脑白质病变;其中,患者1白质稀薄更突出,患者2主要表现为白质萎缩、脑室扩大。发病年龄是VWM严重程度的临床预测因子,起病越早则病情进展越迅速。应激是发病及神经恶化的诱因,目前VWM尚无有效的治疗方法。该文对上述2个病例进行报道,旨在提升临床医师对疾病的认识及早期诊断的能力,以期延缓疾病的进展、延长患者的生存期。
文摘Mutations in the eIF2B gene cause the VWM disease. Genetic and biochemical data of MS patient and MRI data showing VWM images similar to MS lesions, encouraged the present study in which we analyzed the eIF2B5 gene in 225 unrelated MS patients to evaluate an overlapping between MS and VWM. A common variation Ile587Val was found very frequent in the MS patients respect normal controls, thus suggesting that Ile587Val should be considered as susceptibility factor in the development of MS. In conclusion, our data strongly highlight a possible involvement of the eIF2B5 in the development of MS.
文摘Eukaryotic translation initiation factor eIF2B, the guanine nucleotide exchange factor (GEF) for eIF2, catalyzes conversion of eIF2·GDP to eIF2·GTP. Since translation initiation can
文摘In translation initiation, eIF2 is responsible for delivering initiator tRNA to ribosome, and such delivery only occurs when GTP is bound to eIF2. eIF2B, a guanine nucleotide
基金This work was supported by the National Programs for High Technology Research and Development Program(863 Program)(Grant No.2006AA02A316)the National Basic Research Program(973 Program)(Grant Nos.2004CB520801,2006CB910903,2007CB914304,2009CB825501 and 2010CB912301)+1 种基金the Ministry of Science and Technology,National Natural Science Foundation of China(Grant Nos.30721003 and 30870484)the Chinese Academy of Sciences(Grant No.KSCX2-YW-R61).
文摘Eukaryotic translation initiation factor eIF2B,the guanine nucleotide exchange factor(GEF)for eIF2,catalyzes conversion of eIF2·GDP to eIF2·GTP.The eIF2B is composed of five subunits,α,β,γ,δandε,within which theεsubunit is responsible for catalyzing the guanine exchange reaction.Here we present the crystal structure of the C-terminal domain of human eIF2Bε(eIF2Bε-CTD)at 2.0-Åresolution.The structure resembles a HEAT motif and three charge-rich areas on its surface can be identified.When compared to yeast eIF2Bε-CTD,one area involves highly conserved AA boxes while the other two are only partially conserved.In addition,the previously reported mutations in human eIF2Bε-CTD,which are related to the loss of the GEF activity and human VWM disease,have been discussed.Based on the structure,most of such mutations tend to destabilize the HEAT motif.
文摘自质消融性自质脑病（leukoencephalopathy with vanishing white matter，VWM）又称儿童共济失调伴中枢神经系统髓鞘化不良（childhood ataxia with central nervous system hypomyelination，CACH，OMIM306896），是一种常染色体隐性遗传的向质脑病，是儿童遗传性自质脑病中常见的类型之一，其患病率尚缺乏统计。
基金grants from the Natural Science Foundation of China，National Key Technology R and D Program，Key Laboratory Program of Ministry of Education
文摘Vanishing white matter disease （VWM）, a human atitosomal recessive inherited leukoencephalopathy, is due to mutations in eukaryotic initiation factor 2B （elF2B）. elF2B is responsible for tile initiation of protein synthesis by its guanine nucleotide exchange lhctor （GEF） activity. Mutations ofelF2B impair GEF activity at different degree. Previous studies implied improperly activated unlblded protein response （UPR） and endoplasmic reticulum stress （ERS） participated in the pathogenesis ofVWM. Autophagy relieves endoplasmic reticulum load by eliminating the unfolded protein. It is still unknown the effects of genotypes on the pathogenesis. In this work, UPR and autophagy flux were analyzed with different mutational types. Methods： ERS tolerance, reflected by apoptosis and cell viability, was detected in human oligodendrocyte cell line transfected with the wild type, or different mutations of p. Argl 13 His, p. Arg269＊ or p. Ser610-Asp613del in el F2 Be. A representative U PR-PERK component of activating transcription lhctor 4 （ATF4） was measured under the basal condition and ERS induction. Autophagy was analyzed the flux in the presence of lysosomal inhibitors. Results： The degree of ERS tolerance varied in different genotypes. The truncated or deletion mutant showed prominent apoptosis cell viability declination after ERS induction. The most seriously damaged GEF activity ofp. Arg269＊ group underwent spontaneous apoptosis. The truncated or deletion mutant showed elevated ATF4 under basal as well as ERS condition. Decreased expression of LC3-1 and LC3-11 in the mutants reflected an impaired autophagy flux, which was more obvious in the truncated or deletion mutants alter ERS induction. Conclusions： GEF activities in dilt;erent genotypes could influence the cell ERS tolerance as well as compensatory pathways of UPR and autophagy. Oligodendrocytes with truncated or deletion inutants showed less tolerable to ERS.
