【目的】本研究旨在扩增丽江猪清道夫受体B族成员2(scavenger receptor class B member 2,SCARB2)基因序列并进行生物信息学分析,检测其在丽江猪各组织中的表达情况,为后续探究该基因的功能提供理论依据。【方法】采集6月龄丽江猪脂肪组...【目的】本研究旨在扩增丽江猪清道夫受体B族成员2(scavenger receptor class B member 2,SCARB2)基因序列并进行生物信息学分析,检测其在丽江猪各组织中的表达情况,为后续探究该基因的功能提供理论依据。【方法】采集6月龄丽江猪脂肪组织,参考GenBank中野猪(Sus scrofa)的SCARB2基因mRNA序列(登录号:NM_001244155.1)设计引物扩增SCARB2基因CDS区序列并测序,利用生物信息学软件分析SCARB2基因的序列特征和密码子偏好性,分析多个物种与丽江猪、中外猪种与丽江猪SCARB2基因序列的相似性并构建系统进化树,预测SCARB2蛋白理化性质和蛋白结构。使用实时荧光定量PCR检测SCARB2基因在2、4、6月龄丽江猪和6月龄杜洛克猪背部脂肪、心脏、肝脏、肺脏、肾脏等组织中的表达情况。【结果】丽江猪SCARB2基因CDS区序列总长为1437 bp,编码478个氨基酸。丽江猪与山羊、绵羊、牛、马、大鼠、小鼠、人和原鸡的相似性分别为91.3%、91.2%、91.0%、89.8%、82.6%、82.5%、76.0%和66.0%;在6个地方猪种中,除巴马小型猪外,丽江猪与中国地方猪的相似性均为99.4%,与外种猪的相似性为99.1%~99.3%。系统进化树分析显示,丽江猪与野猪、牛、山羊、绵羊聚为一支,原鸡形成单独分支。丽江猪SCARB2蛋白属于亲水蛋白,有2个跨膜螺旋和1个信号肽切割位点,包含10个N-糖基化位点和39个磷酸化位点;其二级结构与三级结构均以无规则卷曲为主。SCARB2基因在6月龄丽江猪背部脂肪和肝脏组织中的表达量极显著高于其他组织(P<0.01);随着月龄增长,SCARB2基因在丽江猪腹部脂肪、背部脂肪和肩部脂肪组织中的表达量逐渐升高,且在2、6月龄间达显著或极显著水平(P<0.05;P<0.01);6月龄丽江猪背部脂肪组织中SCARB2基因表达量极显著高于杜洛克猪(P<0.01)。【结论】本研究成功扩增了丽江猪SCARB2基因序列并分析了其分子特征,其在皮下脂肪、肝脏和肺脏等多个组织中均有表达,且随月龄增长在皮下脂肪组织中的表达量呈显著增加。因此,SCARB2可作为研究丽江猪脂肪沉积性状的候选基因,研究结果可为进一步探究猪脂肪沉积的分子机制提供参考。展开更多
目的系统评价清道夫受体B1基因(SCARB1)变异与心血管疾病(CVD)发病风险的相关性。方法计算机检索PubMed、Web of Science、CNKI、WanFang Data和VIP数据库,检索时限均从建库截至2017年12月31日。由两位研究者独立筛选文献、提取资料和...目的系统评价清道夫受体B1基因(SCARB1)变异与心血管疾病(CVD)发病风险的相关性。方法计算机检索PubMed、Web of Science、CNKI、WanFang Data和VIP数据库,检索时限均从建库截至2017年12月31日。由两位研究者独立筛选文献、提取资料和评价纳入研究的偏倚风险后,采用Stata 12.0软件进行Meta分析。结果共纳入12个研究,报告了SCARB1基因rs5888 C/T多态性、rs4238001 G/A多态性和rs10846744G/C多态性与心血管疾病易感性的相关性。Meta分析结果显示,rs10846744 G/C多态性与心血管疾病发病风险相关[G vs. C:OR=1.30,95%CI(1.11,1.52),P=0.001],但rs5888 C/T多态性[C vs. T:OR=0.97,95%CI(0.86,1.09),P=0.627]和rs4238001 G/A多态性[G vs. A:OR=0.87,95%CI(0.64,1.17),P=0.344]与心血管疾病发病风险不相关。亚组分析结果显示:rs5888 C/T多态性与非亚洲人群CVD的发病风险相关[C vs. T:OR=0.82,95%CI(0.68,0.99),P=0.040]。结论 SCARB1 rs10846744 G/C多态性与CVD发病风险相关。但受纳入研究的数量和质量的限制,上述结论尚待开展更多高质量研究予以验证。展开更多
清道夫受体B类成员1(scavenger receptor class B member 1,Scarb1)作为细胞表面的膜受体蛋白,在动物体色形成过程中发挥重要作用。为了解Scarb1基因在虹鳟(Oncorhynchus mykiss)体色形成中的作用,通过RACE技术克隆虹鳟Scarb1基因的cDN...清道夫受体B类成员1(scavenger receptor class B member 1,Scarb1)作为细胞表面的膜受体蛋白,在动物体色形成过程中发挥重要作用。为了解Scarb1基因在虹鳟(Oncorhynchus mykiss)体色形成中的作用,通过RACE技术克隆虹鳟Scarb1基因的cDNA全长,并运用生物信息学方法分析该基因及其序列结构特征,同时使用实时定量PCR(qRT-PCR)检测Scarb1基因在虹鳟、金鳟及其杂交F_(1)代不同发育阶段和不同组织中的表达情况。结果显示,Scarb1基因cDNA序列全长为2032 bp,开放阅读框1479 bp,编码492个氨基酸,预测分子质量为55.59 ku,且存在保守的CD36结构域和2个跨膜区。序列同源性分析显示,虹鳟与其他硬骨鱼类的氨基酸序列相似度为71.69%~98.58%;进化分析发现虹鳟与大马哈鱼亲缘关系最近,与哺乳动物和两栖动物亲缘关系最远。qRT-PCR检测结果表明,在虹鳟与金鳟胚胎期及出膜后各发育阶段中Scarb1基因均有不同程度表达,且表现为受精期至桑葚期的表达显著高于其他时期(P<0.05),对虹鳟与金鳟同一时期的差异分析发现该基因在胚胎期及7 dph(days post hatch)、1 M(month post hatch)、2 M和3 M时期中表达存在显著差异(P<0.01)。Scarb1基因在虹鳟与金鳟背部皮肤和背部肌肉等色素沉着性组织中表达量较高,其中在金鳟背部皮肤的表达量显著高于虹鳟(P<0.01)。此外,Scarb1基因在杂交F_(1)代不同发育时期中的表达规律与双亲一致;在不同组织中,该基因在杂交F_(1)代背部皮肤中的表达量介于双亲之间。研究结果表明,Scarb1基因与虹鳟体色形成有着密切关系,且可能在金鳟黄色体色形成过程中发挥重要作用。展开更多
The autotetraploid Carassius auratus(4nRR,4n=200,RRRR)is derived from whole-genome duplication of Carassius auratus red var.(RCC,2n=100,RR).In the current study,we demonstrated that chromatophores and pigment changes ...The autotetraploid Carassius auratus(4nRR,4n=200,RRRR)is derived from whole-genome duplication of Carassius auratus red var.(RCC,2n=100,RR).In the current study,we demonstrated that chromatophores and pigment changes directly caused the coloration and variation of 4nRR skin(red in RCC,brownish-yellow in4nRR).To further explore the molecular mechanisms underlying coloration formation and variation in 4nRR,we performed transcriptome profiling and molecular functional verification in RCC and 4nRR.Results revealed that scarb1,associated with carotenoid metabolism,underwent significant down-regulation in 4nRR.Efficient editing of this candidate pigment gene provided clear evidence of its significant role in RCC coloration.Subsequently,we identified four divergent scarb1 homeologs in 4nRR:two original scarb1 homeologs from RCC and two duplicated ones.Notably,three of these homeologs possessed two highly conserved alleles,exhibiting biased and allelespecific expression in the skin.Remarkably,after precise editing of both the original and duplicated scarb1homeologs and/or alleles,4nRR individuals,whether singly or multiply mutated,displayed a transition from brownishyellow skin to a cyan-gray phenotype.Concurrently,the proportional areas of the cyan-gray regions displayed a gene-dose correlation.These findings illustrate the subfunctionalization of duplicated scarb1,with all scarb1genes synergistically and equally contributing to the pigmentation of 4nRR.This is the first report concerning the functional differentiation of duplicated homeologs in an autopolyploidfish,substantiallyenrichingour understanding of coloration formation and change within this group of organisms.展开更多
Unlike the well-established picture for the entry of enveloped viruses, the mechanism of cellular entry of non-enveloped eukaryotic viruses remains largely mysterious. Picornaviruses are representative models for such...Unlike the well-established picture for the entry of enveloped viruses, the mechanism of cellular entry of non-enveloped eukaryotic viruses remains largely mysterious. Picornaviruses are representative models for such viruses, and initiate this entry process by their functional receptors. Here we present the structural and functional studies of SCARB2, a functional receptor of the important human enterovirus 71 （EV71）. SCARB2 is responsible for attachment as well as uncoating of EV71. Differences in the structures of SCARB2 under neutral and acidic conditions reveal that SCARB2 undergoes a pivotal pH-dependent conformational change which opens a lipid-transfer tunnel to mediate the expulsion of a hydrophobic pocket factor from the virion, a pre-requisite for uncoating. We have also identified the key residues essential for attachment to SCARB2, identifying the canyon region of EV71 as mediating the receptor interaction. Together these results provide a clear understanding of cellular attachment and initiation of uncoating for enteroviruses.展开更多
Entero virus 71 （EV71） causes hand, foot, and mouth disease （HFMD） and occasionally leads to severe neurological complications and even death. Scavenger receptor class B member 2 （SCARB2） is a functional recepto...Entero virus 71 （EV71） causes hand, foot, and mouth disease （HFMD） and occasionally leads to severe neurological complications and even death. Scavenger receptor class B member 2 （SCARB2） is a functional receptor for EV71, that mediates viral attachment, internalization, and uncoating. However, the exact binding site of EV71 on SCARB2 is unknown. In this study, we generated a monoclonal antibody （mAb） that binds to human but not mouse SCARB2. It is named JL2, and it can effectively inhibit EV71 infection of target cells. Using a set of chimeras of human and mouse SCARB2, we identified that the region containing residues 77-113 of human SCARB2 contributes significantly to JL2 binding. The structure of the SCARB2-JL2 complex revealed that JL2 binds to the apical region of SCARB2 involving a-helices 2, 5, and 14. Our results provide new insights into the potential binding sites for EV71 on SCARB2 and the molecular mechanism of EV71 entry.展开更多
Background:Angiogenesis is described as a complex process in which new microvessels sprout from endothelial cells of existing vasculature.This study aimed to determine whether long non-coding RNA(lncRNA)H19 induced th...Background:Angiogenesis is described as a complex process in which new microvessels sprout from endothelial cells of existing vasculature.This study aimed to determine whether long non-coding RNA(lncRNA)H19 induced the angiogenesis of gastric cancer(GC)and its possible mechanism.Methods:Gene expression level was determined by quantitative real-time polymerase chain reaction and western blotting.Cell counting kit-8,transwell,5-Ethynyl-2′-deoxyuridine(EdU),colony formation assay,and human umbilical vein endothelial cells(HUVECs)angiogenesis assay as well as Matrigel plug assay were conducted to study the proliferation,migration,and angiogenesis of GC in vitro and in vivo.The binding protein of H19 was found by RNA pull-down and RNA Immunoprecipitation(RIP).High-throughput sequencing was performed and next Gene Ontology(GO)as well as Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis was conducted to analyze the genes that are under H19 regulation.Methylated RIP(me-RIP)assay was used to investigate the sites and abundance among target mRNA.The transcription factor acted as upstream of H19 was determined through chromatin immunoprecipitation(ChIP)and luciferase assay.Results:In this study,we found that hypoxia-induced factor(HIF)-1a could bind to the promoter region of H19,leading to H19 overexpression.High expression of H19 was correlated with angiogenesis in GC,and H19 knocking down could inhibit cell proliferation,migration and angiogenesis.Mechanistically,the oncogenic role of H19 was achieved by binding with the N^(6)-methyladenosine(m^(6)A)reader YTH domain-containing family protein 1(YTHDF1),which could recognize the m^(6)A site on the 3′-untransated regions(3′-UTR)of scavenger receptor class B member 1(SCARB1)mRNA,resulting in over-translation of SCARB1 and thus promoting the proliferation,migration,and angiogenesis of GC cells.Conclusion:HIF-1a induced overexpression of H19 via binding with the promoter of H19,and H19 promoted GC cells proliferation,migration and angiogenesis through YTHDF1/SCARB1,which might be a beneficial target for antiangiogenic therapy for GC.展开更多
Background： Progressive myoclonus epilepsies （PMEs） conaprise a group of rare genetic disorders characterized by action rnyoclonus, epileptic seizures, and ataxia with progressive neurologic decline. Due to clinica...Background： Progressive myoclonus epilepsies （PMEs） conaprise a group of rare genetic disorders characterized by action rnyoclonus, epileptic seizures, and ataxia with progressive neurologic decline. Due to clinical and genetic heterogeneity of PMEs, it is difficult to decide which genes are affected. The aim of this study was to report an action myoclonus with or without renal failure syndrome （EPM4） fhmily and summarize the clinical and genetic characteristics of all reported EPM4 patients. Meihods： In the present study, targeted next-generation sequencing （NGS） was applied to screen causative genes in a Chinese PME family. The candidaie variant was further confirmed by cosegregation analysis and further functional analysis, including the reverse transcription polymerase chain reaction and Western blot of the proband＇s muscle. Moreovel, literature data on the clinical and mutational features of all reported EPM4 patients were reviewed. Results： The gene analysis revealed a novel homozygous splicing mutation （c.995-1G〉A） of the SCARB2 gene in two brothers. Further functional analysis revealed that this mutation led to loss function of the SCARB2 protein. The classification of the candidate variant, according to the American College of Medical Genetics and Genomics standards and guidelines and functional analysis, was pathogenic. Therefore, these two brothers were finally diagnostically confirmed as EPM4. Conclusions： These present results suggest the potential for targeted NGS to conduct a more rapid and precise diagnosis for PME patients. A literature review revealed that mutations in the different functional domains of SCARB2 appear to be associated with the phenotype of EPM4.展开更多
Porcine reproductive and respiratory syndrome(PRRS),a highly infectious immunosuppressive disease caused by porcine reproductive and respiratory syndrome virus(PRRSV),has led to significant economic losses in the glob...Porcine reproductive and respiratory syndrome(PRRS),a highly infectious immunosuppressive disease caused by porcine reproductive and respiratory syndrome virus(PRRSV),has led to significant economic losses in the global swine industry.The complexity of preventing and controlling PRRS,compounded by the limited efficacy of current vaccines,underscores the urgent need to identify antiviral targets and develop effective therapeutics against PRRSV.From the perspective of virus-host interactions,the discovery of target molecules associated with PRRSV resistance offers a promising strategy for future disease management.In this study,we conduct a comprehensive proteomic analysis using data-independent acquisition(DIA)mode to investigate the host response throughout the acute phase of PRRSV infection.This approach provides critical insights into the regulation of host antiviral and immune pathways during acute infection,advancing our theoretical understanding of PRRSV-host interactions and host gene dynamics during this critical phase.Notably,we identified SCARB2,a major lysosomal membrane protein associated with cholesterol metabolism,as a potential regulator of PRRSV replication.These findings offer novel perspectives for the prevention and control of PRRSV,contributing to the development of targeted antiviral strategies.展开更多
文摘【目的】本研究旨在扩增丽江猪清道夫受体B族成员2(scavenger receptor class B member 2,SCARB2)基因序列并进行生物信息学分析,检测其在丽江猪各组织中的表达情况,为后续探究该基因的功能提供理论依据。【方法】采集6月龄丽江猪脂肪组织,参考GenBank中野猪(Sus scrofa)的SCARB2基因mRNA序列(登录号:NM_001244155.1)设计引物扩增SCARB2基因CDS区序列并测序,利用生物信息学软件分析SCARB2基因的序列特征和密码子偏好性,分析多个物种与丽江猪、中外猪种与丽江猪SCARB2基因序列的相似性并构建系统进化树,预测SCARB2蛋白理化性质和蛋白结构。使用实时荧光定量PCR检测SCARB2基因在2、4、6月龄丽江猪和6月龄杜洛克猪背部脂肪、心脏、肝脏、肺脏、肾脏等组织中的表达情况。【结果】丽江猪SCARB2基因CDS区序列总长为1437 bp,编码478个氨基酸。丽江猪与山羊、绵羊、牛、马、大鼠、小鼠、人和原鸡的相似性分别为91.3%、91.2%、91.0%、89.8%、82.6%、82.5%、76.0%和66.0%;在6个地方猪种中,除巴马小型猪外,丽江猪与中国地方猪的相似性均为99.4%,与外种猪的相似性为99.1%~99.3%。系统进化树分析显示,丽江猪与野猪、牛、山羊、绵羊聚为一支,原鸡形成单独分支。丽江猪SCARB2蛋白属于亲水蛋白,有2个跨膜螺旋和1个信号肽切割位点,包含10个N-糖基化位点和39个磷酸化位点;其二级结构与三级结构均以无规则卷曲为主。SCARB2基因在6月龄丽江猪背部脂肪和肝脏组织中的表达量极显著高于其他组织(P<0.01);随着月龄增长,SCARB2基因在丽江猪腹部脂肪、背部脂肪和肩部脂肪组织中的表达量逐渐升高,且在2、6月龄间达显著或极显著水平(P<0.05;P<0.01);6月龄丽江猪背部脂肪组织中SCARB2基因表达量极显著高于杜洛克猪(P<0.01)。【结论】本研究成功扩增了丽江猪SCARB2基因序列并分析了其分子特征,其在皮下脂肪、肝脏和肺脏等多个组织中均有表达,且随月龄增长在皮下脂肪组织中的表达量呈显著增加。因此,SCARB2可作为研究丽江猪脂肪沉积性状的候选基因,研究结果可为进一步探究猪脂肪沉积的分子机制提供参考。
文摘目的系统评价清道夫受体B1基因(SCARB1)变异与心血管疾病(CVD)发病风险的相关性。方法计算机检索PubMed、Web of Science、CNKI、WanFang Data和VIP数据库,检索时限均从建库截至2017年12月31日。由两位研究者独立筛选文献、提取资料和评价纳入研究的偏倚风险后,采用Stata 12.0软件进行Meta分析。结果共纳入12个研究,报告了SCARB1基因rs5888 C/T多态性、rs4238001 G/A多态性和rs10846744G/C多态性与心血管疾病易感性的相关性。Meta分析结果显示,rs10846744 G/C多态性与心血管疾病发病风险相关[G vs. C:OR=1.30,95%CI(1.11,1.52),P=0.001],但rs5888 C/T多态性[C vs. T:OR=0.97,95%CI(0.86,1.09),P=0.627]和rs4238001 G/A多态性[G vs. A:OR=0.87,95%CI(0.64,1.17),P=0.344]与心血管疾病发病风险不相关。亚组分析结果显示:rs5888 C/T多态性与非亚洲人群CVD的发病风险相关[C vs. T:OR=0.82,95%CI(0.68,0.99),P=0.040]。结论 SCARB1 rs10846744 G/C多态性与CVD发病风险相关。但受纳入研究的数量和质量的限制,上述结论尚待开展更多高质量研究予以验证。
文摘清道夫受体B类成员1(scavenger receptor class B member 1,Scarb1)作为细胞表面的膜受体蛋白,在动物体色形成过程中发挥重要作用。为了解Scarb1基因在虹鳟(Oncorhynchus mykiss)体色形成中的作用,通过RACE技术克隆虹鳟Scarb1基因的cDNA全长,并运用生物信息学方法分析该基因及其序列结构特征,同时使用实时定量PCR(qRT-PCR)检测Scarb1基因在虹鳟、金鳟及其杂交F_(1)代不同发育阶段和不同组织中的表达情况。结果显示,Scarb1基因cDNA序列全长为2032 bp,开放阅读框1479 bp,编码492个氨基酸,预测分子质量为55.59 ku,且存在保守的CD36结构域和2个跨膜区。序列同源性分析显示,虹鳟与其他硬骨鱼类的氨基酸序列相似度为71.69%~98.58%;进化分析发现虹鳟与大马哈鱼亲缘关系最近,与哺乳动物和两栖动物亲缘关系最远。qRT-PCR检测结果表明,在虹鳟与金鳟胚胎期及出膜后各发育阶段中Scarb1基因均有不同程度表达,且表现为受精期至桑葚期的表达显著高于其他时期(P<0.05),对虹鳟与金鳟同一时期的差异分析发现该基因在胚胎期及7 dph(days post hatch)、1 M(month post hatch)、2 M和3 M时期中表达存在显著差异(P<0.01)。Scarb1基因在虹鳟与金鳟背部皮肤和背部肌肉等色素沉着性组织中表达量较高,其中在金鳟背部皮肤的表达量显著高于虹鳟(P<0.01)。此外,Scarb1基因在杂交F_(1)代不同发育时期中的表达规律与双亲一致;在不同组织中,该基因在杂交F_(1)代背部皮肤中的表达量介于双亲之间。研究结果表明,Scarb1基因与虹鳟体色形成有着密切关系,且可能在金鳟黄色体色形成过程中发挥重要作用。
基金supported by the National Natural Science Foundation of China (32172972,U19A2040)Science and Technology Innovation Program of Hunan Province (2021RC4028)+4 种基金Earmarked Fund for China Agriculture Research System (CARS-45)Hunan Provincial Science and Technology Department (2019RS5001)Special Funds for Construction of Innovative Provinces in Hunan Province (2021NK1010)Special Science Found of Nansha-South China Agricultural University Fishery Research Institute,Guangzhou (NSYYKY202305,NSYYKY202306)Aid Program for Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province。
文摘The autotetraploid Carassius auratus(4nRR,4n=200,RRRR)is derived from whole-genome duplication of Carassius auratus red var.(RCC,2n=100,RR).In the current study,we demonstrated that chromatophores and pigment changes directly caused the coloration and variation of 4nRR skin(red in RCC,brownish-yellow in4nRR).To further explore the molecular mechanisms underlying coloration formation and variation in 4nRR,we performed transcriptome profiling and molecular functional verification in RCC and 4nRR.Results revealed that scarb1,associated with carotenoid metabolism,underwent significant down-regulation in 4nRR.Efficient editing of this candidate pigment gene provided clear evidence of its significant role in RCC coloration.Subsequently,we identified four divergent scarb1 homeologs in 4nRR:two original scarb1 homeologs from RCC and two duplicated ones.Notably,three of these homeologs possessed two highly conserved alleles,exhibiting biased and allelespecific expression in the skin.Remarkably,after precise editing of both the original and duplicated scarb1homeologs and/or alleles,4nRR individuals,whether singly or multiply mutated,displayed a transition from brownishyellow skin to a cyan-gray phenotype.Concurrently,the proportional areas of the cyan-gray regions displayed a gene-dose correlation.These findings illustrate the subfunctionalization of duplicated scarb1,with all scarb1genes synergistically and equally contributing to the pigmentation of 4nRR.This is the first report concerning the functional differentiation of duplicated homeologs in an autopolyploidfish,substantiallyenrichingour understanding of coloration formation and change within this group of organisms.
基金ACKNOWLEDGEMENTS We thank Neil Shaw, Haitao Yang, Fei Sun, Yuguang Zhao, Jingshan Ren, David I. Stuart and Elizabeth E. Fry for assistance with the manuscript and advice, Wei Peng, Pi Liu, Jialong Zhang provided expert assistance. We gratefully acknowledge the assistance of the staff of the beamline BL5A and BL17A at the Photon Factory （PF） in Japan with the X-ray diffraction data col- lection. We also thank Core Facility in the Institute of Biophysics, Chinese Academy of Sciences （CAS）. Work was supported by the National Basic Research Program （973 Program） （No. 2014CB542800）, National Natural Science Foundation of China （Grant No. 81330036） and the Strategic Priority Research Program of the Chinese Academy of Sciences, （Grant No. XDB08020200）.
文摘Unlike the well-established picture for the entry of enveloped viruses, the mechanism of cellular entry of non-enveloped eukaryotic viruses remains largely mysterious. Picornaviruses are representative models for such viruses, and initiate this entry process by their functional receptors. Here we present the structural and functional studies of SCARB2, a functional receptor of the important human enterovirus 71 （EV71）. SCARB2 is responsible for attachment as well as uncoating of EV71. Differences in the structures of SCARB2 under neutral and acidic conditions reveal that SCARB2 undergoes a pivotal pH-dependent conformational change which opens a lipid-transfer tunnel to mediate the expulsion of a hydrophobic pocket factor from the virion, a pre-requisite for uncoating. We have also identified the key residues essential for attachment to SCARB2, identifying the canyon region of EV71 as mediating the receptor interaction. Together these results provide a clear understanding of cellular attachment and initiation of uncoating for enteroviruses.
基金This work was supported in part by grants from the National Natural Science Foundation of China （Grant Nos. 31670924 and 31570717）, the National Basic Research Program （973 Program） （No. 2014CB542800） and the Open Research Project （2015kf04） from the State Key Laboratory of Biological Macromolecules of the Institute of Biophysics at the Chinese Academy of Sciences. X. Wang was supported by the Young Elite Scientist Sponsorship by CAST.
文摘Entero virus 71 （EV71） causes hand, foot, and mouth disease （HFMD） and occasionally leads to severe neurological complications and even death. Scavenger receptor class B member 2 （SCARB2） is a functional receptor for EV71, that mediates viral attachment, internalization, and uncoating. However, the exact binding site of EV71 on SCARB2 is unknown. In this study, we generated a monoclonal antibody （mAb） that binds to human but not mouse SCARB2. It is named JL2, and it can effectively inhibit EV71 infection of target cells. Using a set of chimeras of human and mouse SCARB2, we identified that the region containing residues 77-113 of human SCARB2 contributes significantly to JL2 binding. The structure of the SCARB2-JL2 complex revealed that JL2 binds to the apical region of SCARB2 involving a-helices 2, 5, and 14. Our results provide new insights into the potential binding sites for EV71 on SCARB2 and the molecular mechanism of EV71 entry.
文摘Background:Angiogenesis is described as a complex process in which new microvessels sprout from endothelial cells of existing vasculature.This study aimed to determine whether long non-coding RNA(lncRNA)H19 induced the angiogenesis of gastric cancer(GC)and its possible mechanism.Methods:Gene expression level was determined by quantitative real-time polymerase chain reaction and western blotting.Cell counting kit-8,transwell,5-Ethynyl-2′-deoxyuridine(EdU),colony formation assay,and human umbilical vein endothelial cells(HUVECs)angiogenesis assay as well as Matrigel plug assay were conducted to study the proliferation,migration,and angiogenesis of GC in vitro and in vivo.The binding protein of H19 was found by RNA pull-down and RNA Immunoprecipitation(RIP).High-throughput sequencing was performed and next Gene Ontology(GO)as well as Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis was conducted to analyze the genes that are under H19 regulation.Methylated RIP(me-RIP)assay was used to investigate the sites and abundance among target mRNA.The transcription factor acted as upstream of H19 was determined through chromatin immunoprecipitation(ChIP)and luciferase assay.Results:In this study,we found that hypoxia-induced factor(HIF)-1a could bind to the promoter region of H19,leading to H19 overexpression.High expression of H19 was correlated with angiogenesis in GC,and H19 knocking down could inhibit cell proliferation,migration and angiogenesis.Mechanistically,the oncogenic role of H19 was achieved by binding with the N^(6)-methyladenosine(m^(6)A)reader YTH domain-containing family protein 1(YTHDF1),which could recognize the m^(6)A site on the 3′-untransated regions(3′-UTR)of scavenger receptor class B member 1(SCARB1)mRNA,resulting in over-translation of SCARB1 and thus promoting the proliferation,migration,and angiogenesis of GC cells.Conclusion:HIF-1a induced overexpression of H19 via binding with the promoter of H19,and H19 promoted GC cells proliferation,migration and angiogenesis through YTHDF1/SCARB1,which might be a beneficial target for antiangiogenic therapy for GC.
基金This work was supported by the grants from the National Natural Science Foundation of China （No. U1505222, No, 81322017, No. 81500980, and No. 81571100） and the National Key Clinical Specialty Discipline Construction Program and Key Clinical Specialty Discipline Construction Program of Fujian.
文摘Background： Progressive myoclonus epilepsies （PMEs） conaprise a group of rare genetic disorders characterized by action rnyoclonus, epileptic seizures, and ataxia with progressive neurologic decline. Due to clinical and genetic heterogeneity of PMEs, it is difficult to decide which genes are affected. The aim of this study was to report an action myoclonus with or without renal failure syndrome （EPM4） fhmily and summarize the clinical and genetic characteristics of all reported EPM4 patients. Meihods： In the present study, targeted next-generation sequencing （NGS） was applied to screen causative genes in a Chinese PME family. The candidaie variant was further confirmed by cosegregation analysis and further functional analysis, including the reverse transcription polymerase chain reaction and Western blot of the proband＇s muscle. Moreovel, literature data on the clinical and mutational features of all reported EPM4 patients were reviewed. Results： The gene analysis revealed a novel homozygous splicing mutation （c.995-1G〉A） of the SCARB2 gene in two brothers. Further functional analysis revealed that this mutation led to loss function of the SCARB2 protein. The classification of the candidate variant, according to the American College of Medical Genetics and Genomics standards and guidelines and functional analysis, was pathogenic. Therefore, these two brothers were finally diagnostically confirmed as EPM4. Conclusions： These present results suggest the potential for targeted NGS to conduct a more rapid and precise diagnosis for PME patients. A literature review revealed that mutations in the different functional domains of SCARB2 appear to be associated with the phenotype of EPM4.
基金supported by the National Natural Science Foundation of China(grant no.:3217190296,82102755 and 32302887)Guangdong Basic and Applied Basic Research Foundation(grant no.:2023A1515012623 and 2019B1515210030)China Postdoctoral Science Foundation(grant no.:2021M703739)。
文摘Porcine reproductive and respiratory syndrome(PRRS),a highly infectious immunosuppressive disease caused by porcine reproductive and respiratory syndrome virus(PRRSV),has led to significant economic losses in the global swine industry.The complexity of preventing and controlling PRRS,compounded by the limited efficacy of current vaccines,underscores the urgent need to identify antiviral targets and develop effective therapeutics against PRRSV.From the perspective of virus-host interactions,the discovery of target molecules associated with PRRSV resistance offers a promising strategy for future disease management.In this study,we conduct a comprehensive proteomic analysis using data-independent acquisition(DIA)mode to investigate the host response throughout the acute phase of PRRSV infection.This approach provides critical insights into the regulation of host antiviral and immune pathways during acute infection,advancing our theoretical understanding of PRRSV-host interactions and host gene dynamics during this critical phase.Notably,we identified SCARB2,a major lysosomal membrane protein associated with cholesterol metabolism,as a potential regulator of PRRSV replication.These findings offer novel perspectives for the prevention and control of PRRSV,contributing to the development of targeted antiviral strategies.
