In eukaryotes,microtubule polymers are essential for cellular plasticity and fate decisions.End-binding(EB)proteins serve as scaffolds for orchestrating microtubule polymer dynamics and are essential for cellular dyna...In eukaryotes,microtubule polymers are essential for cellular plasticity and fate decisions.End-binding(EB)proteins serve as scaffolds for orchestrating microtubule polymer dynamics and are essential for cellular dynamics and chromosome segregation in mitosis.Here,we show that EB1 forms molecular condensates with TIP150 and MCAK through liquid–liquid phase separation to compartmentalize the kinetochore–microtubule plus-end machinery,ensuring accurate kinetochore–microtubule interactions during chromosome segregation in mitosis.Perturbation of EB1–TIP150 polymer formation by a competing peptide prevents phase separation of the EB1-mediated complex and chromosome alignment at the metaphase equator in both cultured cells and Drosophila embryos.Lys220 of EB1 is dynamically acetylated by p300/CBP-associated factor in early mitosis,and persistent acetylation at Lys220 attenuates phase separation of the EB1-mediated complex,dissolves droplets in vitro,and harnesses accurate chromosome segregation.Our data suggest a novel framework for understanding the organization and regulation of eukaryotic spindle for accurate chromosome segregation in mitosis.展开更多
EB1(the end-binding protein 1)蛋白家族是一群广泛存在且高度保守的微管相关蛋白,存在于从酵母到人类的广泛的生物体中.它与微管正极和中心体结合,参与了绝大部分基于微管的生理过程,包括:维持细胞极性,调节染色体稳定性,有丝分裂纺...EB1(the end-binding protein 1)蛋白家族是一群广泛存在且高度保守的微管相关蛋白,存在于从酵母到人类的广泛的生物体中.它与微管正极和中心体结合,参与了绝大部分基于微管的生理过程,包括:维持细胞极性,调节染色体稳定性,有丝分裂纺锤体的定位,将微管锚定到成核位点.自从1995年,Su等人在人细胞中发现了EB1基因,各种生物体中EB1的同源物质被相继报道.十年来,人们通过对不同生物体的研究,试图揭开EB1在细胞中的分布以及它的生理功能.然而到目前为止,对于EB1的了解还非常有限.本文结合国外的研究成果,对EB1蛋白在调节微管动态、纺锤体定位和染色体的稳定性方面以及它与APC(the adenomatous polyposis coli)之间的相互作用作以综述.展开更多
目的探讨微管末端结合蛋白1(end-binding protein 1,EBl)在小鼠神经系统中表达分布以及在细胞自噬蛋白降解通路中的作用。方法以1.5月龄C57BL/6J野生型小鼠为材料,使用蛋白质印迹法检测EB1在小鼠眼、脑、脊髓和坐骨神经中的表达情况。...目的探讨微管末端结合蛋白1(end-binding protein 1,EBl)在小鼠神经系统中表达分布以及在细胞自噬蛋白降解通路中的作用。方法以1.5月龄C57BL/6J野生型小鼠为材料,使用蛋白质印迹法检测EB1在小鼠眼、脑、脊髓和坐骨神经中的表达情况。以小鼠成纤维细胞为材料,使用免疫荧光和激光共聚焦显微镜技术检测EB1在小鼠成纤维细胞中表达和定位。以HEK293细胞为材料,使用siRNA转染和Western blot法检测EB1缺失对于HEK293细胞自噬通路中关键蛋白的影响。结果微管正端示踪蛋白EB1在小鼠神经系统中表达广泛,且与脊髓比较,坐骨神经中含量显著下降。小鼠成纤维细胞中EB1定位于微管正端,呈彗星样分布。HEK293细胞中敲减EB1蛋白可引起自噬通路中p62和LC3B-Ⅱ蛋白表达水平显著提高(P<0.05),而LC3B-Ⅰ蛋白含量未发生明显变化(P>0.05)。结论EB1在神经系统广泛表达,但具有组织部位差异性。EB1聚集于微管正端,其含量下降可能通过破坏自噬小体的形成和运输引起自噬通路异常。展开更多
Angiogenesis,a process by which the preexisting blood vasculature gives rise to new capillary vessels,is associated with a variety of physiologic and pathologic conditions.However,the molecular mechanism underlying th...Angiogenesis,a process by which the preexisting blood vasculature gives rise to new capillary vessels,is associated with a variety of physiologic and pathologic conditions.However,the molecular mechanism underlying this important process remains poorly understood.Here we show that histone deacetylase 6(HDAC6),a microtubule-associated enzyme critical for cell motility,contributes to angiogenesis by regulating the polarization and migration of vascular endothelial cells.Inhibition of HDAC6 activity impairs the formation of new blood vessels in chick embryos and in angioreactors implanted in mice.The requirement for HDAC6 in angiogenesis is corroborated in vitro by analysis of endothelial tube formation and capillary sprouting.Our data further show that HDAC6 stimulates membrane ruffling at the leading edge to promote cell polarization.In addition,microtubule end binding protein 1(EB1)is important for HDAC6 to exert its activity towards the migration of endothelial cells and generation of capillary-like structures.These results thus identify HDAC6 as a novel player in the angiogenic process and offer novel insights into the molecular mechanism governing endothelial cell migration and angiogenesis.展开更多
目的探讨末端结合蛋白1(end-binding protein 1,EB1)在宫颈组织中的表达及其对自噬标志蛋白LC3和p62/SQSTM1蛋白表达的影响。方法免疫组织化学检测EB1在宫颈癌、宫颈息肉和慢性宫颈炎组织中的表达;siRNA干扰方法抑制EB1基因表达,并采用...目的探讨末端结合蛋白1(end-binding protein 1,EB1)在宫颈组织中的表达及其对自噬标志蛋白LC3和p62/SQSTM1蛋白表达的影响。方法免疫组织化学检测EB1在宫颈癌、宫颈息肉和慢性宫颈炎组织中的表达;siRNA干扰方法抑制EB1基因表达,并采用实时荧光定量PCR和Western blot方法检测此时宫颈癌Siha细胞自噬标志蛋白LC3和p62/SQSTM1基因的表达。结果 EB1的表达部位主要集中在细胞核外周的胞质中。EB1在宫颈癌和宫颈息肉中的阳性表达率显著低于宫颈慢性炎性组织(P<0.05),但在阳性表达的宫颈癌组织中,绝大多数间质细胞表现为阴性;EB1在中、低分化宫颈鳞癌组织中的阳性表达率显著低于高分化宫颈鳞癌组织(P<0.05)。当EB1基因表达在宫颈癌Siha细胞中的表达被抑制时,无论是在mRNA还是在蛋白水平,自噬标志蛋白p62/SQSTM1和LC3的表达均显著增加(P<0.05)。结论 EB1在自噬发生较低的宫颈癌组织中阳性表达率低于自噬发生较高的宫颈慢性炎性组织;抑制EB1在宫颈癌Siha细胞中的表达,则自噬标志蛋白p62/SQSTM1和LC3表达改变,这一发现充分证明EB1在宫颈癌的自噬过程中发挥着一定的生物学效应。展开更多
被人广为熟知的移民美国方式是EB5投资移民,但在美国移民的种类中,EB1人才移民才是最受美国政府欢迎的移民类型。美国EB1-C(Managers and Executive Transferees,简称EB1-C)企业高管移民属于美国职业移民第一优先类别,允许跨国公司将其...被人广为熟知的移民美国方式是EB5投资移民,但在美国移民的种类中,EB1人才移民才是最受美国政府欢迎的移民类型。美国EB1-C(Managers and Executive Transferees,简称EB1-C)企业高管移民属于美国职业移民第一优先类别,允许跨国公司将其在美国境外的高级管理人员派遣到美国的分支机构(或总部)继续从事高级管理工作。EB1-C移民要求在美国的公司与在美境外的外国公司存在一种被认可的跨国公司合作关系。中国母公司或美国以外任何国家的企业体(公司、合伙或独资企业均可),和美国的一个企业体已有或将要建立跨国企业关系,即可为其高级管理人员申请EB1-C移民。展开更多
Objective Colon cancer is a type of cancer with high morbidity and mortality,of which adenocarcinoma is the most common type.Numerous studies have found that long noncoding RNAs(lncRNAs)are related to the occurrence a...Objective Colon cancer is a type of cancer with high morbidity and mortality,of which adenocarcinoma is the most common type.Numerous studies have found that long noncoding RNAs(lncRNAs)are related to the occurrence and development of colon cancer.Autophagy is a key metabolic process in the human body and has a role in affecting cancer growth.In this study,our aim was to explore the correlation between lncRNAs and colon adenocarcinoma(COAD)from the perspective of autophagy.Methods A series of bioinformatics methods were used to explore the correlation between lncRNA and COAD from the perspective of autophagy.Results Four autophagy-related lncRNAs related to the prognosis of COAD were identified:EB1-AS1,LINC02381,AC011462.4,and AC016876.1.These four lncRNAs may act as oncogenes involved in the occurrence and development of COAD.The prognostic model was established,and the accuracy of the model was verified by the receiver operating characteristic curve.The risk score of the model could independently predict the prognosis of patients and was preferable to other clinical indicators,with higher values indicating a worse prognosis of the patients.Gene Set Enrichment Analysis was performed for these four lncRNAs,which showed that the high expression group of these were enriched in the basal cell carcinoma pathway.To make it more convenient for clinicians to use,we constructed a nomogram based on age and risk score,which can be used to evaluate the one-,three-,and five-year survival rates of patients.Conclusion These results can help us understand the mechanism of action of lncRNA on COAD from the perspective of autophagy and may provide new directions for the diagnosis and treatment of COAD.The EB1-AS1 gene in this study is a potential candidate biological target for COAD treatment in the future.展开更多
基金supported by grants from the National Key Research and Development Program of China(2022YFA1303100,2022YFA0806800,2022YFA1302700,and 2017YFA0503600)the National Natural Science Foundation of China(32090040,92153302,92254302,92253305,31621002,21922706,92059102,and 92253301)+1 种基金the Plans for Major Provincial Science&Technology Projects of Anhui Province(202303a0702003),the Ministry of Education(IRT_17R102)the Fundamental Research Funds for the Central Universities(KB9100000007 and KB9100000013).
文摘In eukaryotes,microtubule polymers are essential for cellular plasticity and fate decisions.End-binding(EB)proteins serve as scaffolds for orchestrating microtubule polymer dynamics and are essential for cellular dynamics and chromosome segregation in mitosis.Here,we show that EB1 forms molecular condensates with TIP150 and MCAK through liquid–liquid phase separation to compartmentalize the kinetochore–microtubule plus-end machinery,ensuring accurate kinetochore–microtubule interactions during chromosome segregation in mitosis.Perturbation of EB1–TIP150 polymer formation by a competing peptide prevents phase separation of the EB1-mediated complex and chromosome alignment at the metaphase equator in both cultured cells and Drosophila embryos.Lys220 of EB1 is dynamically acetylated by p300/CBP-associated factor in early mitosis,and persistent acetylation at Lys220 attenuates phase separation of the EB1-mediated complex,dissolves droplets in vitro,and harnesses accurate chromosome segregation.Our data suggest a novel framework for understanding the organization and regulation of eukaryotic spindle for accurate chromosome segregation in mitosis.
文摘目的探讨微管末端结合蛋白1(end-binding protein 1,EBl)在小鼠神经系统中表达分布以及在细胞自噬蛋白降解通路中的作用。方法以1.5月龄C57BL/6J野生型小鼠为材料,使用蛋白质印迹法检测EB1在小鼠眼、脑、脊髓和坐骨神经中的表达情况。以小鼠成纤维细胞为材料,使用免疫荧光和激光共聚焦显微镜技术检测EB1在小鼠成纤维细胞中表达和定位。以HEK293细胞为材料,使用siRNA转染和Western blot法检测EB1缺失对于HEK293细胞自噬通路中关键蛋白的影响。结果微管正端示踪蛋白EB1在小鼠神经系统中表达广泛,且与脊髓比较,坐骨神经中含量显著下降。小鼠成纤维细胞中EB1定位于微管正端,呈彗星样分布。HEK293细胞中敲减EB1蛋白可引起自噬通路中p62和LC3B-Ⅱ蛋白表达水平显著提高(P<0.05),而LC3B-Ⅰ蛋白含量未发生明显变化(P>0.05)。结论EB1在神经系统广泛表达,但具有组织部位差异性。EB1聚集于微管正端,其含量下降可能通过破坏自噬小体的形成和运输引起自噬通路异常。
基金the National Natural Science Foundation of China(Grant Nos.30825022 and 90913021)the Fok Ying Tung Education Foundation(Grant No.111036)the National Basic Research Program of China(Grant No.2007CB914802).
文摘Angiogenesis,a process by which the preexisting blood vasculature gives rise to new capillary vessels,is associated with a variety of physiologic and pathologic conditions.However,the molecular mechanism underlying this important process remains poorly understood.Here we show that histone deacetylase 6(HDAC6),a microtubule-associated enzyme critical for cell motility,contributes to angiogenesis by regulating the polarization and migration of vascular endothelial cells.Inhibition of HDAC6 activity impairs the formation of new blood vessels in chick embryos and in angioreactors implanted in mice.The requirement for HDAC6 in angiogenesis is corroborated in vitro by analysis of endothelial tube formation and capillary sprouting.Our data further show that HDAC6 stimulates membrane ruffling at the leading edge to promote cell polarization.In addition,microtubule end binding protein 1(EB1)is important for HDAC6 to exert its activity towards the migration of endothelial cells and generation of capillary-like structures.These results thus identify HDAC6 as a novel player in the angiogenic process and offer novel insights into the molecular mechanism governing endothelial cell migration and angiogenesis.
文摘被人广为熟知的移民美国方式是EB5投资移民,但在美国移民的种类中,EB1人才移民才是最受美国政府欢迎的移民类型。美国EB1-C(Managers and Executive Transferees,简称EB1-C)企业高管移民属于美国职业移民第一优先类别,允许跨国公司将其在美国境外的高级管理人员派遣到美国的分支机构(或总部)继续从事高级管理工作。EB1-C移民要求在美国的公司与在美境外的外国公司存在一种被认可的跨国公司合作关系。中国母公司或美国以外任何国家的企业体(公司、合伙或独资企业均可),和美国的一个企业体已有或将要建立跨国企业关系,即可为其高级管理人员申请EB1-C移民。
文摘Objective Colon cancer is a type of cancer with high morbidity and mortality,of which adenocarcinoma is the most common type.Numerous studies have found that long noncoding RNAs(lncRNAs)are related to the occurrence and development of colon cancer.Autophagy is a key metabolic process in the human body and has a role in affecting cancer growth.In this study,our aim was to explore the correlation between lncRNAs and colon adenocarcinoma(COAD)from the perspective of autophagy.Methods A series of bioinformatics methods were used to explore the correlation between lncRNA and COAD from the perspective of autophagy.Results Four autophagy-related lncRNAs related to the prognosis of COAD were identified:EB1-AS1,LINC02381,AC011462.4,and AC016876.1.These four lncRNAs may act as oncogenes involved in the occurrence and development of COAD.The prognostic model was established,and the accuracy of the model was verified by the receiver operating characteristic curve.The risk score of the model could independently predict the prognosis of patients and was preferable to other clinical indicators,with higher values indicating a worse prognosis of the patients.Gene Set Enrichment Analysis was performed for these four lncRNAs,which showed that the high expression group of these were enriched in the basal cell carcinoma pathway.To make it more convenient for clinicians to use,we constructed a nomogram based on age and risk score,which can be used to evaluate the one-,three-,and five-year survival rates of patients.Conclusion These results can help us understand the mechanism of action of lncRNA on COAD from the perspective of autophagy and may provide new directions for the diagnosis and treatment of COAD.The EB1-AS1 gene in this study is a potential candidate biological target for COAD treatment in the future.