Cilium,an organelle with a unique proteome and organization,protruding from the cell surface,generally serves as a force generator and signaling compartment.During ciliogenesis,ciliary proteins are synthesized in cyto...Cilium,an organelle with a unique proteome and organization,protruding from the cell surface,generally serves as a force generator and signaling compartment.During ciliogenesis,ciliary proteins are synthesized in cytoplasm and transported into cilia by intraflagellar transport(IFT)particles,where the inner counterparts undergo reverse trafficking.The homeostasis of IFT plays a key role in cilial structure assembly and signaling transduction.Much progress has been made on the mechanisms and functions of IFT;however,recent studies have revealed the involvement of IFT particle subunits in organogenesis and spermatogenesis.In this review,we discuss new concepts concerning the molecular functions of IFT protein IFT25 and how its interactions with other IFT particle subunits are involved in mammalian development and fertility.展开更多
目的:研究纤毛转运蛋白140(intraflagellar transport protein 140,IFT140)阳性细胞在小鼠生长发育过程中的动态分布与转归。方法:利用Cre/loxP系统构建IFT140谱系示踪小鼠动物模型,将构建的IFT140-CreER小鼠与R26RtdTomato小鼠进行交...目的:研究纤毛转运蛋白140(intraflagellar transport protein 140,IFT140)阳性细胞在小鼠生长发育过程中的动态分布与转归。方法:利用Cre/loxP系统构建IFT140谱系示踪小鼠动物模型,将构建的IFT140-CreER小鼠与R26RtdTomato小鼠进行交配繁殖,获得基因型为IFT140-CreER;R26RtdTomato小鼠。用他莫昔芬(tamoxifen)腹腔注射诱导Cre重组酶表达,并在特定的时间点观察IFT140阳性细胞的分布、分化及转归规律。结果:聚合酶链反应(polymerase chain reaction,PCR)验证小鼠基因型符合IFT140-CreER;R26RtdTomato。结果显示,阳性细胞在纤毛富集的组织中信号清晰,模型构建成功;骨组织中的IFT140阳性细胞首先出现在骨髓中,随着骨骼发育分布于骨小梁及骨皮质表面。结论:本研究成功构建了谱系示踪小鼠模型,为观察IFT140在小鼠体内重要器官尤其是骨组织发育中的作用,提供实验动物模型。展开更多
纤毛内转运蛋白20(intraflagellar transport protein 20,IFT20)是细胞纤毛内转运蛋白B类中的一种,定位于高尔基体,在纤毛内蛋白转运、纤毛组装和维持纤毛正常形态等多种生理功能的调节中具有关键性作用。IFT20的异常表达与肿瘤、多囊...纤毛内转运蛋白20(intraflagellar transport protein 20,IFT20)是细胞纤毛内转运蛋白B类中的一种,定位于高尔基体,在纤毛内蛋白转运、纤毛组装和维持纤毛正常形态等多种生理功能的调节中具有关键性作用。IFT20的异常表达与肿瘤、多囊肾、免疫紊乱等多种疾病的发生有关,可能是一些重大疾病防治的分子靶点。展开更多
CD4^(+)T cells play major roles in the adaptive immune system,which requires antigen recognition,costimulation,and cytokines for its elaborate orchestration.Recent studies have provided new insight into the importance...CD4^(+)T cells play major roles in the adaptive immune system,which requires antigen recognition,costimulation,and cytokines for its elaborate orchestration.Recent studies have provided new insight into the importance of the supramolecular activation cluster(SMAC),which comprises concentric circles and is involved in the amplification of CD4^(+)T cell activation.However,the underlying mechanism of SMAC formation remains poorly understood.Here,we performed single-cell RNA sequencing of CD4^(+)T cells left unstimulated and stimulated with anti-CD3 and anti-CD28 antibodies to identify novel proteins involved in their regulation.We found that intraflagellar transport 20(IFT20),previously known as cilia-forming protein,was upregulated in antibody-stimulated CD4^(+)T cells compared to unstimulated CD4^(+)T cells.We also found that IFT20 interacted with tumor susceptibility gene 101(TSG101),a protein that endocytoses ubiquitinated T-cell receptors.The interaction between IFT20 and TSG101 promoted SMAC formation,which led to amplification of AKT-mTOR signaling.However,IFT20-deficient CD4^(+)T cells showed SMAC malformation,resulting in reduced CD4^(+)T cell proliferation,aerobic glycolysis,and cellular respiration.Finally,mice with T-cell-specific IFT20 deficiency exhibited reduced allergen-induced airway inflammation.Thus,our data suggest that the IFT20-TSG101 axis regulates AKT-mTOR signaling via SMAC formation.展开更多
目的 :探讨纤毛内运输(intral agellar transport,IFT)蛋白20在肺癌组织中的表达及IFT20对肺癌A549细胞增殖的影响。方法 :免疫组织化学法检测20例肺癌组织和4例正常肺组织中IFT20蛋白的表达。将靶向IFT20基因的小干扰RNA(small interfe...目的 :探讨纤毛内运输(intral agellar transport,IFT)蛋白20在肺癌组织中的表达及IFT20对肺癌A549细胞增殖的影响。方法 :免疫组织化学法检测20例肺癌组织和4例正常肺组织中IFT20蛋白的表达。将靶向IFT20基因的小干扰RNA(small interfering RNA,si RNA)片段转染肺癌A549细胞后,实时荧光定量PCR法检测细胞中IF T20 m RNA的表达,MTT法检测细胞的增殖情况,免疫荧光染色法检测A549细胞中IFT20蛋白的表达情况以及纤毛的数量和长度,蛋白芯片检测A549细胞中蛋白的表达情况。结果 :IFT20蛋白在肺癌组织中呈弱阳性表达,而在正常肺组织中呈中度阳性表达。IFT20 si RNA成功转染后,A549细胞中IFT20 m RNA表达水平低于阴性对照组(转染control si RNA)和空白对照组(未转染的A549细胞)(P<0.05),细胞增殖加快(P<0.05);IF T20 si RNA转染组A549细胞IFT20蛋白表达水平下调,纤毛数量减少、长度变短或消失(P值均<0.05);IFT20 si RNA转染组A549细胞中X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)、survivin、高温需求因子A(high temperature requirment A,HTRA)蛋白、热休克蛋白(heat shock protein,Hsp)27、Hsp70和SMAC(second mitochondria-derived activator of caspases)的表达水平上调(P值均<0.05)。结论 :在肺癌组织中存在IFT20蛋白的低表达,抑制IFT20可以使肺癌A549细胞的纤毛数量减少和纤毛长度变短或消失,并促进肺癌细胞的增殖。展开更多
基金supported by the Natural Science Foundation of Hubei Province(Excellent Youth Foundation,No.2018CFA040)the Outstanding Young and Middle-aged Technology Innovation Team Project of the Hubei Provincial Department of Education(No.T2020003)+1 种基金NIH grant(No.HD076257)Wayne State University Start-up fund.
文摘Cilium,an organelle with a unique proteome and organization,protruding from the cell surface,generally serves as a force generator and signaling compartment.During ciliogenesis,ciliary proteins are synthesized in cytoplasm and transported into cilia by intraflagellar transport(IFT)particles,where the inner counterparts undergo reverse trafficking.The homeostasis of IFT plays a key role in cilial structure assembly and signaling transduction.Much progress has been made on the mechanisms and functions of IFT;however,recent studies have revealed the involvement of IFT particle subunits in organogenesis and spermatogenesis.In this review,we discuss new concepts concerning the molecular functions of IFT protein IFT25 and how its interactions with other IFT particle subunits are involved in mammalian development and fertility.
文摘目的:研究纤毛转运蛋白140(intraflagellar transport protein 140,IFT140)阳性细胞在小鼠生长发育过程中的动态分布与转归。方法:利用Cre/loxP系统构建IFT140谱系示踪小鼠动物模型,将构建的IFT140-CreER小鼠与R26RtdTomato小鼠进行交配繁殖,获得基因型为IFT140-CreER;R26RtdTomato小鼠。用他莫昔芬(tamoxifen)腹腔注射诱导Cre重组酶表达,并在特定的时间点观察IFT140阳性细胞的分布、分化及转归规律。结果:聚合酶链反应(polymerase chain reaction,PCR)验证小鼠基因型符合IFT140-CreER;R26RtdTomato。结果显示,阳性细胞在纤毛富集的组织中信号清晰,模型构建成功;骨组织中的IFT140阳性细胞首先出现在骨髓中,随着骨骼发育分布于骨小梁及骨皮质表面。结论:本研究成功构建了谱系示踪小鼠模型,为观察IFT140在小鼠体内重要器官尤其是骨组织发育中的作用,提供实验动物模型。
文摘纤毛内转运蛋白20(intraflagellar transport protein 20,IFT20)是细胞纤毛内转运蛋白B类中的一种,定位于高尔基体,在纤毛内蛋白转运、纤毛组装和维持纤毛正常形态等多种生理功能的调节中具有关键性作用。IFT20的异常表达与肿瘤、多囊肾、免疫紊乱等多种疾病的发生有关,可能是一些重大疾病防治的分子靶点。
基金the National Research Foundation of Korea(NRF-2021M3A9D3026428)the Ministry of Science and ICT of Korea.J.Jeong is a recipient of funding from the Global Ph.D.Fellowship Program(NRF-2019H1A2A1076865)of the National Research Foundation of Korea.
文摘CD4^(+)T cells play major roles in the adaptive immune system,which requires antigen recognition,costimulation,and cytokines for its elaborate orchestration.Recent studies have provided new insight into the importance of the supramolecular activation cluster(SMAC),which comprises concentric circles and is involved in the amplification of CD4^(+)T cell activation.However,the underlying mechanism of SMAC formation remains poorly understood.Here,we performed single-cell RNA sequencing of CD4^(+)T cells left unstimulated and stimulated with anti-CD3 and anti-CD28 antibodies to identify novel proteins involved in their regulation.We found that intraflagellar transport 20(IFT20),previously known as cilia-forming protein,was upregulated in antibody-stimulated CD4^(+)T cells compared to unstimulated CD4^(+)T cells.We also found that IFT20 interacted with tumor susceptibility gene 101(TSG101),a protein that endocytoses ubiquitinated T-cell receptors.The interaction between IFT20 and TSG101 promoted SMAC formation,which led to amplification of AKT-mTOR signaling.However,IFT20-deficient CD4^(+)T cells showed SMAC malformation,resulting in reduced CD4^(+)T cell proliferation,aerobic glycolysis,and cellular respiration.Finally,mice with T-cell-specific IFT20 deficiency exhibited reduced allergen-induced airway inflammation.Thus,our data suggest that the IFT20-TSG101 axis regulates AKT-mTOR signaling via SMAC formation.
文摘目的 :探讨纤毛内运输(intral agellar transport,IFT)蛋白20在肺癌组织中的表达及IFT20对肺癌A549细胞增殖的影响。方法 :免疫组织化学法检测20例肺癌组织和4例正常肺组织中IFT20蛋白的表达。将靶向IFT20基因的小干扰RNA(small interfering RNA,si RNA)片段转染肺癌A549细胞后,实时荧光定量PCR法检测细胞中IF T20 m RNA的表达,MTT法检测细胞的增殖情况,免疫荧光染色法检测A549细胞中IFT20蛋白的表达情况以及纤毛的数量和长度,蛋白芯片检测A549细胞中蛋白的表达情况。结果 :IFT20蛋白在肺癌组织中呈弱阳性表达,而在正常肺组织中呈中度阳性表达。IFT20 si RNA成功转染后,A549细胞中IFT20 m RNA表达水平低于阴性对照组(转染control si RNA)和空白对照组(未转染的A549细胞)(P<0.05),细胞增殖加快(P<0.05);IF T20 si RNA转染组A549细胞IFT20蛋白表达水平下调,纤毛数量减少、长度变短或消失(P值均<0.05);IFT20 si RNA转染组A549细胞中X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)、survivin、高温需求因子A(high temperature requirment A,HTRA)蛋白、热休克蛋白(heat shock protein,Hsp)27、Hsp70和SMAC(second mitochondria-derived activator of caspases)的表达水平上调(P值均<0.05)。结论 :在肺癌组织中存在IFT20蛋白的低表达,抑制IFT20可以使肺癌A549细胞的纤毛数量减少和纤毛长度变短或消失,并促进肺癌细胞的增殖。
