TMEM43基因编码的跨膜蛋白43(transmembrane protein 43,TMEM43)是TMEM蛋白家族成员之一,该蛋白质约为400个氨基酸,包括4个跨膜结构域和1个膜内结构域。TMEM43在许多物种中都存在表达,并且遗传相似性很高,特别是4个跨膜结构在不同物种...TMEM43基因编码的跨膜蛋白43(transmembrane protein 43,TMEM43)是TMEM蛋白家族成员之一,该蛋白质约为400个氨基酸,包括4个跨膜结构域和1个膜内结构域。TMEM43在许多物种中都存在表达,并且遗传相似性很高,特别是4个跨膜结构在不同物种中都表现为高度保守。近年来有研究者发现,TMEM43与听神经病谱系障碍(auditory neuropathy spectrum disorder,ANSD)的发生可能相关,推测其可能为一种新的听力损伤相关基因。本文就现阶段TMEM43基因与听力损伤的关系展开综述,分析TMEM43在耳发育与声传导方面的作用,探讨TMEM43基因变异对听力损伤产生的影响,以期为TMEM43后续研究和精准医疗提供新思路。展开更多
为探究人源跨膜蛋白43(TMEM43)基因含量、定位分布及体外真核表达情况,利用RT-PCR及Western blot方法分别检测内源MRC-5和HEK293细胞内TMEM43基因及其蛋白表达情况,采用间接免疫荧光实验进一步分析其在MRC-5细胞内的分布;构建重组表达载...为探究人源跨膜蛋白43(TMEM43)基因含量、定位分布及体外真核表达情况,利用RT-PCR及Western blot方法分别检测内源MRC-5和HEK293细胞内TMEM43基因及其蛋白表达情况,采用间接免疫荧光实验进一步分析其在MRC-5细胞内的分布;构建重组表达载体pcDNA3.1-EGFP-TMEM43,转染至HEK293细胞内检测外源TMEM43在HEK293细胞内的基因和蛋白表达情况。结果显示,基因水平TMEM43在MRC-5细胞内的表达明显高于HEK293细胞,蛋白水平仅能检测到MRC-5细胞内TMEM43的表达。重组表达质粒pcDNA3.1-EGFP-TMEM43转染HEK293细胞后,基因水平TMEM43表达显著升高, Western blot可检测到转染后细胞内TMEM43蛋白成功表达。结果表明,不同细胞表达TMEM43含量存在差异,肺来源细胞内TMEM43的表达明显高于肾来源的细胞;构建的重组载体pcDNA3.1-EGFP-TMEM43可成功转染HEK293细胞并表达。该研究为TMEM43结构及功能、与疾病相关机理的进一步探究提供了实验依据。展开更多
Arrhythmogenic right ventricular dysplasia/cardiomyopathy(ARVD/C)is a genetic cardiac muscle disease that accounts for approximately 30%sudden cardiac death in young adults.The Ser358Leu mutation of transmembrane prot...Arrhythmogenic right ventricular dysplasia/cardiomyopathy(ARVD/C)is a genetic cardiac muscle disease that accounts for approximately 30%sudden cardiac death in young adults.The Ser358Leu mutation of transmembrane protein 43(TMEM43)was commonly identified in the patients of highly lethal and fully penetrant ARVD subtype,ARVD5.Here,we generated TMEM43 S358L mouse to explore the underlying mechanism.This mouse strain showed the classic patholo.gies of ARVD patients,including structural abnormalities and cardiac fibrofatty.TMEM43 S358L mutation led to hyper-activated nuclear factor kB(NFkB)activation in heart tissues and primary cardiomy.ocyte cells.Importantly,this hyper activation of NF-κB directly drove the expression of pro-fibrotic gene,transforming growth factor beta(TGFβ),and enhanced downstream signal,indicating that TMEM43 S358L mutation up-regulates NF-κB-TGFβ signal cascade during ARVD cardiac fibrosis.Our study partially reveals the regulatory mechanism of ARVD development.展开更多
目的 探讨跨膜蛋白43(transmembrane protein 43,TMEM43)在脑心肌炎病毒(encephalomyocarditis virus,EMCV)感染过程中的作用。方法 采用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantitation,i TRAQ)结合...目的 探讨跨膜蛋白43(transmembrane protein 43,TMEM43)在脑心肌炎病毒(encephalomyocarditis virus,EMCV)感染过程中的作用。方法 采用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantitation,i TRAQ)结合液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry,LC-MS/MS)高通量蛋白质组学技术,对EMCV感染的BHK-21细胞进行蛋白质组学分析;采用分子克隆技术构建人源TMEM43重组质粒p CMV-MycTMEM43,转染HEK293细胞后,感染EMCV(MOI=3),real-time PCR法检测TMEM43过表达对EMCV体外增殖以及EMCV吸附和进入的影响;RNAi试验筛选最有效干扰序列,转染HEK293细胞后,感染EMCV(MOI=3),real-time PCR法检测TMEM43下调对EMCV体外增殖的影响。结果 EMCV感染的BHK-21细胞TMEM43表达显著增加。重组质粒p CMV-Myc-TMEM43可在HEK293细胞正常表达,TMEM43过表达对EMCV复制有显著促进作用,病毒感染滴度也有一定提高;TMEM43过表达在病毒进入阶段发挥促进作用,而对吸附过程无任何影响。TMEM43下调对EMCV增殖有明显抑制作用。结论 TMEM43促进EMCV复制增殖,主要在病毒进入阶段发挥作用。展开更多
文摘为探究人源跨膜蛋白43(TMEM43)基因含量、定位分布及体外真核表达情况,利用RT-PCR及Western blot方法分别检测内源MRC-5和HEK293细胞内TMEM43基因及其蛋白表达情况,采用间接免疫荧光实验进一步分析其在MRC-5细胞内的分布;构建重组表达载体pcDNA3.1-EGFP-TMEM43,转染至HEK293细胞内检测外源TMEM43在HEK293细胞内的基因和蛋白表达情况。结果显示,基因水平TMEM43在MRC-5细胞内的表达明显高于HEK293细胞,蛋白水平仅能检测到MRC-5细胞内TMEM43的表达。重组表达质粒pcDNA3.1-EGFP-TMEM43转染HEK293细胞后,基因水平TMEM43表达显著升高, Western blot可检测到转染后细胞内TMEM43蛋白成功表达。结果表明,不同细胞表达TMEM43含量存在差异,肺来源细胞内TMEM43的表达明显高于肾来源的细胞;构建的重组载体pcDNA3.1-EGFP-TMEM43可成功转染HEK293细胞并表达。该研究为TMEM43结构及功能、与疾病相关机理的进一步探究提供了实验依据。
基金grants from National Natural Science Foundation of China(81570211 to X.Lin)China Postdoctoral Science Foundation(023221010 to G.Zheng).
文摘Arrhythmogenic right ventricular dysplasia/cardiomyopathy(ARVD/C)is a genetic cardiac muscle disease that accounts for approximately 30%sudden cardiac death in young adults.The Ser358Leu mutation of transmembrane protein 43(TMEM43)was commonly identified in the patients of highly lethal and fully penetrant ARVD subtype,ARVD5.Here,we generated TMEM43 S358L mouse to explore the underlying mechanism.This mouse strain showed the classic patholo.gies of ARVD patients,including structural abnormalities and cardiac fibrofatty.TMEM43 S358L mutation led to hyper-activated nuclear factor kB(NFkB)activation in heart tissues and primary cardiomy.ocyte cells.Importantly,this hyper activation of NF-κB directly drove the expression of pro-fibrotic gene,transforming growth factor beta(TGFβ),and enhanced downstream signal,indicating that TMEM43 S358L mutation up-regulates NF-κB-TGFβ signal cascade during ARVD cardiac fibrosis.Our study partially reveals the regulatory mechanism of ARVD development.
