Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis(Bt)have proven to be highly effective in managing some key pests.However,the evolution of resistance by the target pests threa...Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis(Bt)have proven to be highly effective in managing some key pests.However,the evolution of resistance by the target pests threatens the sustainability of Bt crops.The L31S mutation in a tetraspanin encoded by Harm TspC5(previously known as Ha TSPAN1)has been shown to confer dominant resistance to the Bt protein Cry1Ac in Helicoverpa armigera,a globally damaging lepidopteran pest.However,the broader implications of the L31S mutation in the tetraspanins of other lepidopteran species remain unclear.The evolutionary analyses in this study indicate that TspC5s have evolved in a species-specific manner among the lepidopteran insects.To investigate the role of TspC5s in conferring dominant resistance to Cry1Ac,we used the piggyBac-based transformation system to generate four transgenic H.armigera strains that express exogenous TspC5 variants from three phylogenetically close species(Helicoverpa zea,Helicoverpa assulta and Heliothis virescens)and one phylogenetically distant species(Plutella xylostella).In comparison with the background SCD strain of H.armigera,the transgenic strains expressing HzeaTspC5-L31S,HassTspC5-L31S,or HvirTspC5-L31S exhibited significant resistance to Cry1Ac(10.0-,21.4-,and 81.1-fold,respectively),whereas the strain expressing PxylTspC5-L27S remained susceptible.Furthermore,the Cry1Ac resistant phenotypes followed an autosomal dominant inheritance pattern and were closely linked to the introduced mutant TspC5s.These findings reveal the conserved role of TspC5s from Helicoverpa and Heliothis species in mediating the dominant resistance to Cry1Ac,and they provide crucial insights for assessing resistance risks related to mutant tetraspanins and devising adaptive resistance management strategies for these major lepidopteran pests.展开更多
Objective:To identify a full length cDNA sequence of a novel tetraspanin(TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.M...Objective:To identify a full length cDNA sequence of a novel tetraspanin(TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.Methods:Using the NCBI,EMBI,Expasy and other online sites, the open reading frame(ORF),conserved domain,physical and chemical parameters,signal peptide,transmembrane domain,epitope,topological structures of the protein sequences were predicted.And Vector NTI software was used for multiple sequence alignment and phylogenetic tree construction.Results:’Hie target sequence was 1 132 hp length with a 681 hp biggest ORF encoding 226 amino acids protein with typical TSP conserved domain.It was confirmed as full length cDNA of TSP16 from Spirometra erinaceieuropaei and named as SeTSP16 (GenBank accession number:JF728872).The predicted molecular weight and isoelectric point of the deduced protein were 24 750.5 Da and 7.88 Da,respectively.Compared with TSP16s from Schistosoma japonicum and Schistosoma mansoni.it showed similarity of 59%and 59%, respectively.SeTSP16 contained four transmembrane domains(TM 1-4),intracellular N and C-termini,one short small extracellular loop and one large extracellular loop.Four major epitopes that were significant different from the corresponding epitope regions of TSP16 from Schistosoma mansoni and Schistosoma japonicum were predicted.Conclusions:The full length cDNA sequences of SeTSP16 arc identified.It encodes a transmembrane protein which might be an ideal diagnosis antigen and target molecule for antiparasitic drugs.展开更多
This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specifi...This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin αbβ3 by the two mAbs. Results.HI117 and SJ9A4(10μg/ml and 20μg/ml) induced evident specific Fg binding to human platelets,suggesting that the two mAbs evoked activation of platelet integrin αbβ3.Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αbβ3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. Conclusion.The anti platelet tetraspanin(CD9)mAbs,HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors.Three signaling pathways,i.e.thromboxane,secreted ADP, and cAMP pathways may be involved in the process,with protein kinase C activation presumably being the common step of the three pathways.展开更多
The tetraspanin gene family encodes cell-surface proteins that span the membrane 4 times and play critical roles in a wide range of biological processes across numerous organisms.Recent findings highlight the involvem...The tetraspanin gene family encodes cell-surface proteins that span the membrane 4 times and play critical roles in a wide range of biological processes across numerous organisms.Recent findings highlight the involvement of a tetraspanin of the lepidopteran pest Helicoverpa armigera in resistance to Bacillus thuringiensis Cry insecticidal proteins,which are extensively used in transgenic crops.Thus,a better understanding of lepidopteran tetraspanins is urgently needed.In the current study,genome scanning in 10 lepidopteran species identified a total of 283 sequences encoding potential tetraspanins.Based on conserved cysteine patterns in the large extracellular loop and their phylogenetic relationships,these tetraspanins were classified into 8 subfamilies(TspA to TspH).Six ancestral introns were identified within lepidopteran tetraspanin genes.Tetraspanins in TspA,TspB,TspC,and TspD subfamilies exhibit highly similar gene organization,while tetraspanins in the remaining 4 subfamilies exhibited variation in intron loss and/or gain during evolution.Analysis of chromosomal distribution revealed a lepidopteran-specific cluster of 10 to 11 tetraspanins,likely formed by tandem duplication events.Selective pressure analysis indicated negative selection across all orthologous groups,withωvalues ranging between 0.004 and 0.362.However,positive selection was identified at 18 sites within TspB5,TspC5,TspE3,and TspF10.Furthermore,spatiotemporal expression analysis of H.armigera tetraspanins demonstrated variable expression levels across different developmental stages and tissues,suggesting diverse functions of tetraspanin members in this globally important insect pest.Our findings establish a solid foundation for subsequent functional investigations of tetraspanins in lepidopteran species.展开更多
Cimex species are ectoparasites that exclusively feed on warm-blooded animals such as birds and mammals.Three cimicid species are known to be persistent pests for humans,including the tropical bed bug Cimex hemipterus...Cimex species are ectoparasites that exclusively feed on warm-blooded animals such as birds and mammals.Three cimicid species are known to be persistent pests for humans,including the tropical bed bug Cimex hemipterus,common bed bug Cimex lectularius,and Eastern bat bug Leptocimex boueti.To date,genomic information is restricted to the common bed bug C.lectularius,which limits understanding their biology and to provide controls of bed bug infestations.Here,a chromosomal-level genome assembly of C.hemipterus(495 Mb[megabase pairs])contained on 16 pseudochromosomes(scaffold N50=34 Mb),together with 9 messenger RNA and small RNA transcriptomes were obtained.In comparison between hemipteran genomes,we found that the tetraspanin superfamily was expanded in the Cimex ancestor.This study provides the first genome assembly for the tropical bed bug C.hemipterus,and offers an unprecedented opportunity to address questions relating to bed bug infestations,as well as genomic evolution to hemipterans more widely.展开更多
characterize the activation of platelet integrin αⅡbβ3 induced by two anti human platelet tetraspanin monoclonal antibodies (mAbs), HI117 and SJ9A4, and investigate their potential mechanism of action Method...characterize the activation of platelet integrin αⅡbβ3 induced by two anti human platelet tetraspanin monoclonal antibodies (mAbs), HI117 and SJ9A4, and investigate their potential mechanism of action Methods Using 125 I labeled human fibrinogen (Fg), specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured Results HI117 and SJ9A4 (10?μg/ml and 20?μg/ml) induced specific Fg binding to human platelets, suggesting that the two mAbs evoked activation of platelet integrin αⅡbβ3 Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αⅡbβ3 activation was inhibited by pretreatment of platelets with sphingosine, aspirin, apyrase, and/or PGI 2 Conclusions Anti platelet tetraspanin (CD9) mAbs, HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors Three signaling pathways, namely thromboxane, secreted ADP, and cAMP pathways, may be involved in the process, with protein kinase C activation presumably being the common step of the three pathways展开更多
家蚕(Bombyx mori)是一种重要的特种经济昆虫,由家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)引起的血液型脓病对养蚕业造成了严重危害。四跨膜蛋白(tetraspanin)在细胞信号转导、病毒识别与入侵以及膜融合等多种生理...家蚕(Bombyx mori)是一种重要的特种经济昆虫,由家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)引起的血液型脓病对养蚕业造成了严重危害。四跨膜蛋白(tetraspanin)在细胞信号转导、病毒识别与入侵以及膜融合等多种生理过程中发挥着关键作用,在病毒复制与宿主免疫应答中亦具有重要功能。从家蚕中筛选鉴定出一个潜在的四跨膜蛋白基因,将其命名为Tetraspanin B(BmTsp.B),其编码区序列包含765 bp,共编码254个氨基酸。组织表达谱分析显示BmTsp.B在脂肪体中的转录水平最高,且BmNPV感染后转录水平显著上升。基因过表达和siRNA介导的敲降实验表明,BmTsp.B能够增强BmNPV的感染与复制。qRT-PCR分析结果显示,BmTsp.B能够参与调控凋亡相关基因的表达进而促进病毒对细胞的侵染能力。上述研究结果可为明晰BmTsp.B蛋白的功能以及深入解析BmNPV致病机制提供基础,同时为家蚕抗性品种培育以及BmNPV科学防控提供新的靶标。展开更多
基金primarily funded by a grant from the National Natural Science Foundation of China(31930093)Additional support was provided by the Natural Science Foundation of Jiangsu Province,China(BK20230983)the Project of Fund for Stable Support to Agricultural Sci-Tech Renovation,China(xjnkywdzc-2022004)。
文摘Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis(Bt)have proven to be highly effective in managing some key pests.However,the evolution of resistance by the target pests threatens the sustainability of Bt crops.The L31S mutation in a tetraspanin encoded by Harm TspC5(previously known as Ha TSPAN1)has been shown to confer dominant resistance to the Bt protein Cry1Ac in Helicoverpa armigera,a globally damaging lepidopteran pest.However,the broader implications of the L31S mutation in the tetraspanins of other lepidopteran species remain unclear.The evolutionary analyses in this study indicate that TspC5s have evolved in a species-specific manner among the lepidopteran insects.To investigate the role of TspC5s in conferring dominant resistance to Cry1Ac,we used the piggyBac-based transformation system to generate four transgenic H.armigera strains that express exogenous TspC5 variants from three phylogenetically close species(Helicoverpa zea,Helicoverpa assulta and Heliothis virescens)and one phylogenetically distant species(Plutella xylostella).In comparison with the background SCD strain of H.armigera,the transgenic strains expressing HzeaTspC5-L31S,HassTspC5-L31S,or HvirTspC5-L31S exhibited significant resistance to Cry1Ac(10.0-,21.4-,and 81.1-fold,respectively),whereas the strain expressing PxylTspC5-L27S remained susceptible.Furthermore,the Cry1Ac resistant phenotypes followed an autosomal dominant inheritance pattern and were closely linked to the introduced mutant TspC5s.These findings reveal the conserved role of TspC5s from Helicoverpa and Heliothis species in mediating the dominant resistance to Cry1Ac,and they provide crucial insights for assessing resistance risks related to mutant tetraspanins and devising adaptive resistance management strategies for these major lepidopteran pests.
文摘Objective:To identify a full length cDNA sequence of a novel tetraspanin(TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.Methods:Using the NCBI,EMBI,Expasy and other online sites, the open reading frame(ORF),conserved domain,physical and chemical parameters,signal peptide,transmembrane domain,epitope,topological structures of the protein sequences were predicted.And Vector NTI software was used for multiple sequence alignment and phylogenetic tree construction.Results:’Hie target sequence was 1 132 hp length with a 681 hp biggest ORF encoding 226 amino acids protein with typical TSP conserved domain.It was confirmed as full length cDNA of TSP16 from Spirometra erinaceieuropaei and named as SeTSP16 (GenBank accession number:JF728872).The predicted molecular weight and isoelectric point of the deduced protein were 24 750.5 Da and 7.88 Da,respectively.Compared with TSP16s from Schistosoma japonicum and Schistosoma mansoni.it showed similarity of 59%and 59%, respectively.SeTSP16 contained four transmembrane domains(TM 1-4),intracellular N and C-termini,one short small extracellular loop and one large extracellular loop.Four major epitopes that were significant different from the corresponding epitope regions of TSP16 from Schistosoma mansoni and Schistosoma japonicum were predicted.Conclusions:The full length cDNA sequences of SeTSP16 arc identified.It encodes a transmembrane protein which might be an ideal diagnosis antigen and target molecule for antiparasitic drugs.
文摘This study characterized the activation of platelet integrin α bβ3 induced by two anti human platelet tetraspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods.Using 125 I labeled human fibrinogen(Fg),specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin αbβ3 by the two mAbs. Results.HI117 and SJ9A4(10μg/ml and 20μg/ml) induced evident specific Fg binding to human platelets,suggesting that the two mAbs evoked activation of platelet integrin αbβ3.Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αbβ3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. Conclusion.The anti platelet tetraspanin(CD9)mAbs,HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors.Three signaling pathways,i.e.thromboxane,secreted ADP, and cAMP pathways may be involved in the process,with protein kinase C activation presumably being the common step of the three pathways.
基金funded by the Natural Science Foundation of Jiangsu Province of China(grant no.BK20230983).
文摘The tetraspanin gene family encodes cell-surface proteins that span the membrane 4 times and play critical roles in a wide range of biological processes across numerous organisms.Recent findings highlight the involvement of a tetraspanin of the lepidopteran pest Helicoverpa armigera in resistance to Bacillus thuringiensis Cry insecticidal proteins,which are extensively used in transgenic crops.Thus,a better understanding of lepidopteran tetraspanins is urgently needed.In the current study,genome scanning in 10 lepidopteran species identified a total of 283 sequences encoding potential tetraspanins.Based on conserved cysteine patterns in the large extracellular loop and their phylogenetic relationships,these tetraspanins were classified into 8 subfamilies(TspA to TspH).Six ancestral introns were identified within lepidopteran tetraspanin genes.Tetraspanins in TspA,TspB,TspC,and TspD subfamilies exhibit highly similar gene organization,while tetraspanins in the remaining 4 subfamilies exhibited variation in intron loss and/or gain during evolution.Analysis of chromosomal distribution revealed a lepidopteran-specific cluster of 10 to 11 tetraspanins,likely formed by tandem duplication events.Selective pressure analysis indicated negative selection across all orthologous groups,withωvalues ranging between 0.004 and 0.362.However,positive selection was identified at 18 sites within TspB5,TspC5,TspE3,and TspF10.Furthermore,spatiotemporal expression analysis of H.armigera tetraspanins demonstrated variable expression levels across different developmental stages and tissues,suggesting diverse functions of tetraspanin members in this globally important insect pest.Our findings establish a solid foundation for subsequent functional investigations of tetraspanins in lepidopteran species.
基金chromosome assembly was submitted to NCBI Assembly under accession number JASJUQ000000000 in NCBIgenerated in this study have been deposited into the NCBI database under the BioProject accession PRJNA713496the genome annotation files were deposited in the Figshare(https://figshare.com/s/d9956fa56189ac8938c0).
文摘Cimex species are ectoparasites that exclusively feed on warm-blooded animals such as birds and mammals.Three cimicid species are known to be persistent pests for humans,including the tropical bed bug Cimex hemipterus,common bed bug Cimex lectularius,and Eastern bat bug Leptocimex boueti.To date,genomic information is restricted to the common bed bug C.lectularius,which limits understanding their biology and to provide controls of bed bug infestations.Here,a chromosomal-level genome assembly of C.hemipterus(495 Mb[megabase pairs])contained on 16 pseudochromosomes(scaffold N50=34 Mb),together with 9 messenger RNA and small RNA transcriptomes were obtained.In comparison between hemipteran genomes,we found that the tetraspanin superfamily was expanded in the Cimex ancestor.This study provides the first genome assembly for the tropical bed bug C.hemipterus,and offers an unprecedented opportunity to address questions relating to bed bug infestations,as well as genomic evolution to hemipterans more widely.
基金grantfromtheNationalNaturalScienceFoundationofChina (No 39470 172 )
文摘characterize the activation of platelet integrin αⅡbβ3 induced by two anti human platelet tetraspanin monoclonal antibodies (mAbs), HI117 and SJ9A4, and investigate their potential mechanism of action Methods Using 125 I labeled human fibrinogen (Fg), specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured Results HI117 and SJ9A4 (10?μg/ml and 20?μg/ml) induced specific Fg binding to human platelets, suggesting that the two mAbs evoked activation of platelet integrin αⅡbβ3 Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc receptors, and that HI117 and SJ9A4 induced integrin αⅡbβ3 activation was inhibited by pretreatment of platelets with sphingosine, aspirin, apyrase, and/or PGI 2 Conclusions Anti platelet tetraspanin (CD9) mAbs, HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc receptors Three signaling pathways, namely thromboxane, secreted ADP, and cAMP pathways, may be involved in the process, with protein kinase C activation presumably being the common step of the three pathways
