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| 1 | FKBP8负调控RNA病毒早期诱导的Ⅰ型IFN信号通路显示文摘目的进一步探究VISA介导的抗病毒信号通路。方法利用免疫共沉淀、免疫印迹、双荧光素报告、qPCR等实验技术检测FKBP8在RLRs信号通路中的作用。结果我们的实验证实FKBP8与VISA、TRAF3都存在互作,且FKBP8显著地抑制RNA病毒早期诱导的干扰素启动子IFN-β和核调节因子-κB(nuclear factor kappa-B,NF-κB)的活性,并增强病毒的复制。进一步的研究表明FKBP8通过作用于VISA和TBK1而负调控RLR-VISA信号转导;此外,FKBP8通过干扰VISA-TRAF3的结合而负调节病毒诱导的信号通路。结论FKBP8是RLR抗病毒先天免疫信号通路中的负调控因子。 | 徐杉杉 李圣纳 侯雯 许亮国 | 2020 | 免疫学杂志2020，36，1： | 5 |
| 2 | TRIM35 mediates protection against influenza infection by activating TRAF3 and degrading viral PB2显示文摘Tripartite motif(TRIM)family proteins are important effectors of innate immunity against viral infections.Here we identified TRIM35 as a regulator of TRAF3 activation.Deficiency in or inhibition of TRIM35 suppressed the production of type I interferon(IFN)in response to viral infection.777m35-deficient mice were more susceptible to influenza A virus(IAV)infection than were wild-type mice.TRIM35 promoted the RIG-Imediated signaling by catalyzing Lys63-linked polyubiquitination of TRAF3 and the subsequent formation of a signaling complex with VISA and TBK1.IAV PB2 polymerase countered the innate antiviral immune response by impeding the Lys63-linked polyubiquitination and activation of TRAF3.TRIM35 mediated Lys48-linked polyubiquitination and proteasomal degradation of IAV PB2,thereby antagonizing its suppression of TRAF3 activation.Our in vitro and in vivo findings thus reveal novel roles of TRIM35,through catalyzing Lys63-or Lys48-linked polyubiquitination,in RIG-I antiviral immunity and mechanism of defense against IAV infection. | Nan Sun Li Jiang Miaomiao Ye Yihan Wang Guangwen Wang Xiaopeng Wan Yuhui Zhao Xia Wen Libin Liang Shujie Ma Liling Liu Zhigao Bu Hualan Chen Chengjun Li | 2020 | Protein & Cell2020，11，12： | 4 |
| 3 | 银杏素促进成纤维样滑膜细胞中TRAF3蛋白的表达缓解类风湿关节炎显示文摘目的探究银杏素(ginkgetin)治疗类风湿关节炎(rheumatoid arthritis,RA)的潜在机制。方法通过TNF-α刺激成纤维样滑膜MH7A细胞建立炎性细胞模型,用银杏素单独处理或分别与TNF受体相关因子3(tumor necrosis factor receptor-associated factor 3,TRAF3)的小干扰RNA(si-TRAF3)和p38激动剂茴香霉素共同处理,检测细胞增殖、凋亡、炎症因子白介素(interleukin,IL)-6、IL-1β分泌、TRAF3蛋白表达和p38蛋白磷酸化水平。此外,建立CIA大鼠模型并给予口服银杏素,评估大鼠关节炎指数,足爪肿胀度和滑膜组织病理学,检测滑膜组织TRAF3表达以及TNF-α、IL-6和IL-1β水平。结果银杏素上调了TRAF3表达,抑制了MH7A细胞增殖及炎症因子分泌,促进凋亡,阻断p38 MAPK信号通路。而转染si-TRAF3或添加茴香霉素处理,可逆转银杏素对MH7A细胞增殖、凋亡和炎症因子分泌以及p38 MAPK信号通路的调节作用。此外,银杏素治疗降低了CIA大鼠关节炎指数、足爪肿胀度以及TNF-α,IL-6和IL-1β分泌。结论银杏素能够上调TRAF3表达,阻断p38 MAPK信号通路,抑制MH7A细胞增殖和炎症因子分泌,促进凋亡,进而减轻CIA大鼠的关节症状。 | 吕昌伟 张静涛 郗海涛 强毅 张乾 | 2022 | 中国骨质疏松杂志2022，28，2： | 3 |
| 4 | Influenza A virus use of BinCARD1 to facilitate the binding of viral NP to importinα7 is counteracted by TBK1-p62 axis-mediated autophagy显示文摘As a major component of the viral ribonucleoprotein(vRNP)complex in influenza A virus(IAV),nucleoprotein(NP)interacts with isoforms of importinαfamily members,leading to the import of itself and vRNP complex into the nucleus,a process pivotal in the replication cycle of IAV.In this study,we found that BinCARD1,an isoform of Bcl10-interacting protein with CARD(BinCARD),was leveraged by IAV for efficient viral replication.BinCARD1 promoted the nuclear import of the vRNP complex and newly synthesized NP and thus enhanced vRNP complex activity.Moreover,we found that BinCARD1 interacted with NP to promote NP binding to importinα7,an adaptor in the host nuclear import pathway.However,we also found that BinCARD1 promoted RIG-I-mediated innate immune signaling by mediating Lys63-linked polyubiquitination of TRAF3,and that TBK1 appeared to degrade BinCARD1.We showed that BinCARD1 was polyubiquitinated at residue K103 through a Lys63 linkage,which was recognized by the TBK1-p62 axis for autophagic degradation.Overall,our data demonstrate that IAV leverages BinCARD1 as an important host factor that promotes viral replication,and two mechanisms in the host defense system are triggered—innate immune signaling and autophagic degradation—to mitigate the promoting effect of BinCARD1 on the life cycle of IAV. | Xuyuan Wang Li Jiang Guangwen Wang Wenjun Shi Yuzhen Hu Bo Wang Xianying Zeng Guobin Tian Guohua Deng Jianzhong Shi Liling Liu Chengjun Li Hualan Chen | 2022 | Cellular & Molecular Immunology2022，19，10： | 3 |
| 5 | MicroRNA-628-5p Facilitates Enterovirus 71 Infection by Suppressing TRAF3 Signaling显示文摘To date,EV71 infection still poses a great challenge to the health of infants and young children.1 Much established evidence suggests that EV71 infection depends on a wide variety of host factors,including cell surface receptors for EV71 entry,innate immune response,miRNAs,and lncRNAs.2,3 It is thought that diverse signaling pathways are required for EV71 to escape the host immune response.4 Innate immunity serves as the first line of defense against pathogens.Tumor necrosis factor receptor-associated factor(TRAF)proteins are essential components of signaling pathways activated by Toll-like receptor(TLR)or RIG-I-like receptor(RLR)family members.TRAF3 is widely expressed by many cell types,including all nucleated immune cells,in which it plays many roles in the regulation of immune functions.5 TRAF3 plays a significant role in regulating the functions of B and T lymphocytes.TRAF3 is a highly versatile regulator that positively controls type I interferon and cytokine production through interferon regulatory factors(IRFs)and nuclear factor-κB(NF-κB).6 A recent study indicated that EV71-induced ubiquitin-specific protease 19(USP19)negatively regulates type I IFN signaling by targeting TRAF3. | Dong Li Shuaiyin Chen Weiguo Zhang Chao Zhang Tiantian Sun Yue Du Ronghua Ding Yanlei Gao Yuefei Jin Guangcai Duan | 2021 | Cellular & Molecular Immunology2021，18，5： | 2 |
| 6 | TRAF3和TRAF6在天然免疫中的作用显示文摘近年来,人们发现肿瘤坏死因子受体相关蛋白是肿瘤坏死因子受体(TNF)超家族和白细胞介素-1受体/Toll样受体(IL-1R/TLR)超家族重要信号转导分子。迄今为止,已发现六种不同的TRAFs,分别是TRAF1、TRAF2、TRAF3、TRAF4、TRAF5、TRAF6,其中TRAF3和TRAF6在天然免疫中有重要作用。尽管二者的结构相似,但TRAF3和TRAF6在功能上有很大区别。 | 王桂芹 邓国华 李书华 | 2006 | 畜牧兽医科技信息2006，22，4： | 2 |
| 7 | FXYD3 enhances IL-17A signaling to promote psoriasis by competitively binding TRAF3 in keratinocytes显示文摘Psoriasis is a common chronic inflammatory skin disease characterized by inflammatory cell infiltration and epidermal hyperplasia.However,the regulatory complexity of cytokine and cellular networks still needs to be investigated.Here,we show that the expression of FXYD3,a member of the FXYD domain-containing regulators of Na+/K+ATPases family,is significantly increased in the lesional skin of psoriasis patients and mice with imiquimod(IMQ)-induced psoriasis.IL-17A,a cytokine important for the development of psoriatic lesions,contributes to FXYD3 expression in human primary keratinocytes.FXYD3 deletion in keratinocytes attenuated the psoriasis-like phenotype and inflammation in an IMQ-induced psoriasis model.Importantly,FXYD3 promotes the formation of the IL-17R-ACT1 complex by competing with IL-17R for binding to TRAF3 and then enhances IL-17A signaling in keratinocytes.This promotes the activation of the NF-κB and MAPK signaling pathways and leads to the expression of proinflammatory factors.Our results clarify the mechanism by which FXYD3 serves as a mediator of IL-17A signaling in keratinocytes to form a positive regulatory loop to promote psoriasis exacerbation.Targeting FXYD3 may serve as a potential therapeutic approach in the treatment of psoriasis. | Wenjuan Yang Rukun He Hao Qu Wenwen Lian Yue Xue Tao Wang Wenlong Lin Peishuo Zhu Meng Xia Lihua Lai Qingqing Wang | 2023 | Cellular & Molecular Immunology2023，20，3： | 2 |
| 8 | TRAF3交互蛋白2介导雄性小鼠肥胖相关血管胰岛素抵抗与功能异常显示文摘血管胰岛素抵抗是肥胖的一个特点,可导致血管异常与疾病的发生。然而,目前对肥胖相关的血管胰岛素抵抗及功能异常的潜在分子机制仍了解很少。该文假设TRAF3交互蛋白2(TRAF3 interacting protein 2,TRAF3IP2),一个已知在心血管病激活病理应激通路的促炎症接头分子,与肥胖相关的血管胰岛素抵抗与功能异常有因果联系。研究人员通过体外在内皮细胞、离体动脉中,体内在喂食诱发肥胖的TRAF3IP2敲除小鼠模型中进行基因调控. | 赵狄(摘译) 刘莉(审校) Grunewald ZI Ramirez-Perez FI Woodford ML Morales-Quinones M Mejia S Manrique-Acevedo C Siebenlist U Martinez-Lemus LA Chandrasekar B Padilla J | 2020 | 中华高血压杂志2020，28，12： | 1 |
| 9 | 日本鳗鲡TRAF3基因的克隆及功能研究显示文摘为探究鱼类TRAF3在鱼类抗病毒免疫应答中的功能及作用机制,实验利用逆转录PCR克隆获得了日本鳗鲡TRAF3转录本(AjTRAF3),利用生物信息学软件分析了AjTRAF3的结构特征,利用实时荧光定量PCR(qPCR)、双荧光素酶报告系统以及免疫共沉淀等方法对其表达规律、功能及作用机理进行了初步分析。AjTRAF3的开放阅读框长度为1707 bp,编码568个氨基酸。序列结构分析结果显示,AjTRAF3由N端的环结构域2个锌指结构域以及1个螺旋结构域和C端高度保守的TRAF-C(MATH)结构域组成。qPCR结果显示,AjTRAF3在日本鳗鲡各组织中均有表达,脑组织中表达量最高,其次为头肾,心脏中的表达量最低。Poly I:C刺激6 h后,日本鳗鲡脾脏组织中AjTRAF3上调倍数最高,为对照组的15.83倍。迟缓爱德华氏菌感染24 h后,日本鳗鲡脾脏组织中AjTRAF3上调倍数最高,为对照组的31.47倍。此外,本研究构建了AjTRAF3真核表达质粒,发现过表达AjTRAF3能显著上调炎症及抗病毒相关基因的表达,可显著增强AjIFN2、AjIFN4和NF-κB启动子荧光素酶活性。并能显著上调由AjRIG-IN、AjMAVS、AjIRF3诱导的AjIFN2、AjIFN4和NF-κB启动子活性。免疫荧光结果显示,AjTRAF3主要定位于细胞质中,且与AjMAVS存在共定位。免疫共沉淀结果显示,AjTRAF3通过MATH结构域与AjMAVS相互结合,缺失该结构域后,其与AjMAVS的相互作用消失,推测AjTRAF3可通过介导RIG-I/MAVS信号转导途径调控鱼类的抗病毒免疫应答。本研究结果为进一步揭示鱼类TRAF3的生物学功能奠定了基础。 | 凌露露 梁英 黄文树 聂品 黄贝 | 2023 | 水产学报2023，47，8： | 1 |
| 10 | TRAF3 Interacts with Smac/DIABLO and Enhances the Proapoptotic Effect of Smac/DIABLO in Cytoplasm显示文摘Smac/DIABLO (有低 PI 的 caspase/directIAP-binding 蛋白质的第二导出线粒体的使活跃之物) 是 29 kDa mitochondrial 先锋蛋白质,它 isproteolytically 在线粒体处理了成 23 kDa 成熟蛋白质。它被免除 themitochondrial 内部到在 apoptotic trigger.Smac/DIABLO 以后的 cytosol 的膜空间是行动一更暗淡并且它由扣押贡献 caspase 激活 apoptosis 蛋白质(国际机场) 的禁止者。为了推进,调查 Smac/DIABLO 行动的机制,我们把 Smac/DIABLO 的成熟形式用作一个诱饵并且屏蔽了用酵母在人的 livercDNA 图书馆与成熟 Smac/DIABLO 交往的蛋白质二混血儿的系统。在 5.8x10 ~ 6colonies 被营养限制和 X 牛乳糖试金屏蔽以后, 42 个殖民地被获得。在 DNA 顺序分析和相同检索以后,当 TNFreceptor 的 TRAF 领域联系了因素 3,候选人蛋白质之一被识别(TRAF3 ) 。在 TRAF3 和 Smac/DIABLO 之间的相互作用地点被贝它牛乳糖测试识别。在经由 TRAF 领域的 TRAF3 和 Smac/DIABLO 之间的相互作用被 co-immunoprecipitation 在 HepG2 房间在 vivo 识别,并且在 vitro 的直接相互作用 betweenTRAF3 和 Smac/DIABLO 被GST拉在 TRAF3 的assay.Co表示下面识别,在 293 个房间的成熟 Smac/DIABLO 能提高建议的 Smac/DIABLO-mediated apoptosis.These 结果那 TRAF3 经由 TRAF 领域与 Smac/DIABLO 交往了,在细胞质导致 Smac/DIABLO 的 increasedproapoptotic 效果。 | Cuili ZHANG~(1,2) Lei XIE~4 Hai’en CHENG~3 and Yingxiong WANG~2 ~1Clinical Laboratory,Sichuan Corps Hospital of The people’s Armed Police Forces,Leshan 614000,China ~2Department of Procreation Medicine,Chongqing Medical Universtity,Chongqing 400016,China ~3Clinical Laboratory,Second Affiliated Hospital Zhejiang University College of Medicine,Hangzhou 310009,China ~4Sitchuan Corps Hospital The People’s Armed Police Forces,Leshan 614000,China | 2007 | Acta Biochimica et Biophysica Sinica2007，39，2： | 1 |
| 11 | Profiling of miRNAs in porcine Sertoli cells显示文摘Background: Sertoli cells(SCs) create a specialized environment to support and dictate spermatogenesis.MicroRNAs(miRNAs), a kind of ~ 22 nt small noncoding RNAs, have been reported to be highly abundant in mouse SCs and play critical roles in spermatogenesis. However, the miRNAs of porcine SCs remain largely unknown.Methods: We isolated porcine SCs and conducted small RNA sequencing. By comparing miRNAs in germ cells, we systematically analyzed the miRNA expression pattern of porcine SCs. We screened the highly enriched SC miRNAs and predicted their functions by Gene Ontology analysis. The dual luciferase assay was used to elucidate the regulation of tumor necrosis factor receptor(TNFR)-associated factor 3(TRAF3) by ssc-miR-149.Results: The analysis showed that 18 miRNAs were highly expressed in SCs and 15 miRNAs were highly expressed in germ cells. These miRNAs were predicted to mediate SC and germ cell functions. In addition, ssc-miR-149 played critical roles in SCs by targeting TRAF3.Conclusion: Our findings provide novel insights into the miRNA expression pattern and their regulatory roles of porcine SCs. | Xiaoxu Chen Yi Zheng Xueliang Li Qiang Gao Tongying Feng Pengfei Zhang Mingzhi Liao Xiu’e Tian Hongzhao Lu Wenxian Zeng | 2021 | Journal of Animal Science and Biotechnology2021，12，1： | 1 |
| 12 | 罗非鱼TRAF3基因的表达及功能研究显示文摘肿瘤坏死因子受体(TNFR)相关因子3(TRAF3)是多种免疫途径中的关键调控因子。从尼罗罗非鱼(Oreochromis niloticus)中克隆获得了TRAF3基因,命名为OnTRAF3(GeneBank No.MN258118),该基因包含1个环指结构域、1个锌指结构域、1个卷曲螺旋和MATH结构域。多序列比对表明,OnTRAF3与其他已知的TRAF3蛋白具有高度的相似性,尤其是MATH结构域。实时荧光定量PCR(qRT-PCR)分析显示,OnTRAF3在各组织中广泛分布,且在脑、皮肤、肠和鳃中表达量较高。在无乳链球菌诱导后,多个组织中的OnTRAF3表达量出现了不同程度的上调,说明OnTRAF3参与了罗非鱼的抗菌免疫应答。亚细胞定位实验显示,OnTRAF3分布在HEK293的细胞质和细胞核中。此外,荧光素酶报告基因实验结果显示,野生型(WT)OnTRAF3可显著激活NF-κB信号,而coiled-coil和MATH结构域缺失后,依然能够显著激活NF-κB活性,而RING和Zinc缺失后,该激活作用则明显减弱,表明RING和Zinc结构域是OnTRAF3在免疫信号通路中行使功能的关键结构域。研究为探索TRAF3在罗非鱼免疫应答中的功能提供了重要的基础。 | 夏洪丽 汪志文 黎源 陈文捷 龙梦 喻大鹏 程俊 夏立群 鲁义善 | 2023 | 水生生物学报2023，47，2： | 1 |
| 13 | BAFF-R胞内区与TRAF3相结合关键分子结合域的确定显示文摘目的:研究BAFF-R胞内区与TRAF3相互作用的关键结合域。方法:分别钓取BAFF-R胞内区和TRAF3的c DNA片段,克隆到酵母双杂交系统中,验证两者之间的相互作用。利用缺失PCR和重叠PCR的方法获取11个TRAF3的突变体,验证TRAF3的11个突变体和BAFF-R胞内区的相互作用。结果:TRAF3与BAFF-R胞内区发生相互作用时,有三个关键的分子结合域,分别是N端的螺旋结构382-400位氨基酸、中间的428-463位氨基酸以及TRAF3 C端的543-560位氨基酸。结论:在体内得到了BAFF-R胞内区与TRAF3相互作用的三个关键分子结合域,有助于展开对BAFF-R胞内区与TRAF3相互作用的进一步研究。 | 田侠 徐东刚 杜建芳 | 2015 | 现代生物医学进展2015，15，8： | 0 |
| 14 | 大肠杆菌麦芽糖结合蛋白激活TLR2介导的MyD88依赖途径和TLR4介导的TRIF/TRAF3依赖途径诱导Th1活化显示文摘目的探讨TLR2/TLR4介导的信号通路在大肠杆菌麦芽糖结合蛋白诱导Th1活化中的调控作用。方法免疫磁珠分选方法获取纯的CD4+T细胞,经CD3/CD28抗体活化后的CD4+T细胞分别用MBP、MBP+anti-TLR2、MBP+anti-TLR4刺激。ELISA检测培养上清中IFN-γ和IL-4的分泌;RT-PCR检测CD4+T细胞IFN-γ、MyD88、TRIF、TRAF3和TRAF6的mRNA表达;Western blotting方法检测CD4+T细胞MyD88、TRIF、TRAF3和TRAF6的蛋白表达。结果 MBP组IFN-γ水平升高、IFN-γ、MyD88、TRIF、TRAF3的mRNA上调、TRAF6的mRNA下调;MBP+anti-TLR2组IFN-γ水平降低、TRAF6的mRNA上调、MyD88的mRNA下调、TRIF、TRAF3的mRNA无影响;MBP+anti-TLR4组IFN-γ水平升高、TRAF6的mRNA上调、MyD88、TRIF、TRAF3的mRNA下调,Western blotting实验也出现了相似的结果。结论 TLR2介导的MyD88依赖途径和TLR4介导的TRIF/TRAF3依赖途径在MBP诱导的Th1活化过程中发挥了重要的调控作用。 | 刘果木 翟晓玉 孙敏英 张楠楠 倪伟华 接晶 台桂香 蒋丽娜 | 2017 | 免疫学杂志2017，33，1： | 0 |
| 15 | TRAF3 activates STING-mediated suppression of EV-A71 and target of viral evasion显示文摘Innate immunity represents one of the main host responses to viral infection.1,2,3 STING(Stimulator of interferon genes),a crucial immune adapter functioning in host cells,mediates cGAS(Cyclic GMP-AMP Synthase)sensing of exogenous and endogenous DNA fragments and generates innate immune responses.4 Whether STING activation was involved in infection and replication of enterovirus remains largely unknown.In the present study,we discovered that human enterovirus A71(EV-A71)infection triggered STING activation in a cGAS dependent manner.EV-A71 infection caused mitochondrial damage and the discharge of mitochondrial DNA into the cytosol of infected cells.However,during EV-A71 infection,cGAS-STING activation was attenuated.EV-A71 ^(pro)teins were screened and the viral ^(pro)tease 2A^(pro) had the greatest capacity to inhibit cGAS-STING activation.We identified TRAF3 as an important factor during STING activation and as a target of 2A^(pro).Supplement of TRAF3 rescued cGAS-STING activation suppression by 2A^(pro).TRAF3 supported STING activation mediated TBK1 phosphorylation.Moreover,we found that 2A^(pro) ^(pro)tease activity was essential for inhibiting STING activation.Furthermore,EV-D68 and CV-A16 infection also triggered STING activation.The viral ^(pro)tease 2A^(pro) from EV-D68 and CV-A16 also had the ability to inhibit STING activation.As STING activation prior to EV-A71 infection generated cellular resistance to EV-A71 replication,blocking EV-A71-mediated STING suppression represents a new anti-viral target. | Wenwen Zheng Zhenbang Zhou Yajuan Rui Runxin Ye Fengyan Xia Fei Guo Xiaoman Liu Jiaming Su Meng Lou Xiao-Fang Yu | 2023 | Signal Transduction and Targeted Therapy2023，8，3： | 0 |
| 16 | 脱氢胆酸调控OPG/RANK和TRAF3抑制破骨细胞分化显示文摘目的探讨脱氢胆酸(dehydrocholic acid,DHCA)对破骨细胞(osteoclasts,OCs)分化及功能的影响。方法采用粒细胞-巨噬细胞集落刺激因子和核因子κB受体活化因子配体(receptor activator of nuclear factor kappa B(NF-κB)-ligand,RANKL)诱导成熟骨髓来源的巨噬细胞分化为OCs。通过抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP/ACP5)染色,确定DHCA抑制OCs形成的最佳浓度。qRT-PCR检测OCs分化和功能相关基因TRAP/ACP5、组织蛋白酶K(cathepsin K,CTSK)和基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)的基因表达。Western blot检测OCs中TNF受体相关因子3(TNF receptor-associated factor 3,TRAF3)、NF-κB受体活化因子(receptor activator of NF-κB,RANK)和骨保护素(osteoprotegerin,OPG)的蛋白水平。结果DHCA浓度为200μmol/L是抑制OCs形成的最佳剂量(P<0.01)。DHCA抑制OCs分化和功能相关基因ACP5、CTSK和MMP9的表达(P<0.01)。DHCA通过下调RANK蛋白和增加TRAF3和OPG蛋白的表达来抑制OCs的分化(P<0.01)。结论DHCA通过调控OPG/RANK和TRAF3抑制OCs分化,这可能是一种有效的骨质疏松前体药物。 | 朱禹潼 张晓楠 关溪 尚东 | 2024 | 中国骨质疏松杂志2024，30，1： | 0 |
| 17 | 凋亡蛋白Daxx与TRAF3相互作用的筛选及相互作用位点的鉴定显示文摘目的:利用酵母双杂交技术在成人肝文库中筛选能与死亡结构域相关蛋白(Daxx)相互作用的蛋白,并用哺乳动物细胞中进行了免疫共沉淀验证,以进一步了解Daxx的功能.方法:利用PCR在成人肝cDNA文库中扩增Daxx全长ORF,构建pDBLeu-Daxx载体.转化MaV203,检测细胞毒性和自激活作用,确定His基础表达的3AT浓度.依次转化成人肝cDNA文库.在营养缺陷型培养基上挑取三阳性克隆.进行回转实验和免疫共沉淀验证,并利用β-gal分析进行了相互作用结构域的鉴定.结果:筛库转化效率达到5.8×106个克隆.筛选到了一株能与Daxx相互作用的阳性克隆,DNA序列分析和同源检索表明该阳性克隆为肿瘤坏死因子受体相关因子3(TRAF3).通过回转及co-IP证实了TRAF3能与Daxx相互作用,β-gal分析发现TRAF3通过TRAFDomain与Daxx相互作用.结论:TRAF3通过TRAFDomain与Daxx相互作用,可能参与了Daxx的调控和功能,为进一步了解Daxx作用的分子机制奠定了基础. | 张翠莉 成海恩 王应雄 | 2007 | 第四军医大学学报2007，28，9： | 0 |
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