As an aquatic fish,the spotted halibut Verasper variegatus is highly susceptible to bacterial and virus infections.Tumor necrosis factor-alpha(TNF-α)as a cytokine could control the inflammatory responses.The function...As an aquatic fish,the spotted halibut Verasper variegatus is highly susceptible to bacterial and virus infections.Tumor necrosis factor-alpha(TNF-α)as a cytokine could control the inflammatory responses.The functions of TNF-αin many species have been widely studied,particularly in mammals.However,little is known about the TNF-αfunctions in V.variegatus.We first cloned and sequenced the TNF-αgene in V.variegatus(VvTNF-α).The two conserved cysteine residues,transmembrane sequence,Thr-Leu motif,and TNF family signature,as well as the TA-rich motifs of its proteins related to inflammatory responses had high similarity to those of the other teleost and mammalian TNF-α.The phylogenetic analysis showed that VvTNF-αwas consistent with TNF-αgenes of other vertebrates.The VvTNF-αtranscripts were extensively distributed in the peripheral blood leukocytes(PBLs),spleen,and gill,indicating that the VvTNF-αhad a role in immune function.Furthermore,treatment with pathogen-associated molecular patterns(PAMPs)could induce a rapid and significant increase of VvTNF-αin the PBLs,which reveals that VvTNF-αdoes participate in the host immune responses against bacterial and viral pathogens.We found that VvTNF-αhad an interesting expression pattern during metamorphosis,showing that the flatfish TNF-αmay have some novel functions during specific developmental stages.In addition,the 3 D structure prediction of VvTNF-αprovided an indication of how it is likely to interact with other proteins.Therefore,VvTNF-αhas multiple functions,and provides valuable information to explore novel functions of TNF-α.展开更多
Cold-inducible RNA-binding protein(CIRP) is a kind of RNA binding proteins that plays important roles in many physiological processes. The CIRP has been widely studied in mammals and amphibians since it was first clon...Cold-inducible RNA-binding protein(CIRP) is a kind of RNA binding proteins that plays important roles in many physiological processes. The CIRP has been widely studied in mammals and amphibians since it was first cloned from mammals. On the contrary, there are little reports in teleosts. In this study, the Po CIRP gene of the Japanese flounder was cloned and sequenced. The genomic sequence consists of seven exons and six introns. The putative Po CIRP protein of flounder was 198 amino acid residues long containing the RNA recognition motif(RRM). Phylogenetic analysis showed that the flounder Po CIRP is highly conserved with other teleost CIRPs. The 5' flanking sequence was cloned by genome walking and many transcription factor binding sites were identified. There is a Cp Gs region located in promoter and exon I region and the methylation state is low. Quantitative real-time PCR analysis uncovered that Po CIRP gene was widely expressed in adult tissues with the highest expression level in the ovary. The m RNA of the Po CIRP was maternally deposited and the expression level of the gene was regulated up during the gastrula and neurula stages. In order to gain the information how the protein interacts with m RNA, we performed the modeling of the 3D structure of the flounder Po CIRP. The results showed a cleft existing the surface of the molecular. Taken together, the results indicate that the CIRP is a multifunctional molecular in teleosts and the findings about the structure provide valuable information for understanding the basis of this protein's function.展开更多
Interleukin-1 beta(IL-1β) is a cytokine involved in the control of inflammatory responses.The functions of IL-1β have been extensively studied in many vertebrates.However,few studies focused on the functions of IL-1...Interleukin-1 beta(IL-1β) is a cytokine involved in the control of inflammatory responses.The functions of IL-1β have been extensively studied in many vertebrates.However,few studies focused on the functions of IL-1β in the spotted halibut(Verasper variegatus),an endangered aquatic fish that easily suffers from bacterial and viral infections.We first cloned and sequenced the IL-1β gene from V.variegatus(Vv IL-1β).Its conservative interleukin-1 propeptide and interleukin-1/18 domains corresponding regions,characteristics of the IL-1 protein family and TA-rich motifs relating to inflammatory responses were similar to those of the IL-1β genes from other teleost and mammals.The phylogenetic analysis showed that Vv IL-1β was homologous to the IL-1β gene of other vertebrates.Treatment with pathogen-associated molecular patterns(PAMPs) induced a significant and rapid up-regulation of Vv IL-1β expression in the peripheral blood leukocytes(PBLs),revealing that Vv IL-1β is actively involved in the host immune responses against bacterial and viral pathogens.The expression level of Vv IL-1β was relatively high in the PBLs,gill,and intestine,indicating the immunological function of Vv IL-1β.Vv IL-1β also had interesting expression patterns at specific developmental stages,implying that the IL-1β of flatfishes may have some novel functions during embryonic development and metamorphosis.The 3 D structure prediction also provided information about how Vv IL-1β interacts with its receptor protein.These results suggested that Vv IL-1β may have multiple functions in V.variegatus and provided us valuable information for understanding the novel functions of IL-1β.展开更多
GATA4,as a member of the GATA transcription factor family,plays a significant role in lineage specification and trans-differentiation of cells by regulating target gene expression.In this study,the potential transcrip...GATA4,as a member of the GATA transcription factor family,plays a significant role in lineage specification and trans-differentiation of cells by regulating target gene expression.In this study,the potential transcription factor binding sites in the pro-moter of Paralichthys olivaceus sox2 gene were analyzed,and a GATA4-binding motif was identified.A cleaved amplified poly-morphic sequence-based binding assay experiment was performed,and the results showed a significant binding process between the Homo sapiens GATA4 recombinant protein and the DNA fragment of P.olivaceus sox2 promoter.Luciferase reporter assay revealed that GATA4 could improve the activity of the P.olivaceus sox2 promoter.Overexpression of P.olivaceus GATA4 could also increase the sox2 mRNA expression level in flounder brain cells.Overall,our results indicate that sox2 is a target gene of GATA4 in P.oliva-ceus.Sox2 is implicated in the maintenance of stem cell pluripotency and can reprogram differentiated cells to iPS cells.In addition,sox2 is involved in the development of the central nervous system(CNS).Therefore,this study may provide a reference for exploring the function of P.olivaceus GATA4 in the regulation of pluripotent stem cells and the development of the CNS.展开更多
Marine organisms cannot grow and reproduce without proper metabolic regulation.Within a metabolic network,problems with a given link will affect the normal life activities of the organism.Many metabolic mechanisms ass...Marine organisms cannot grow and reproduce without proper metabolic regulation.Within a metabolic network,problems with a given link will affect the normal life activities of the organism.Many metabolic mechanisms associated with behaviors of Am-phioctopus fangsiao are still unclear.Moreover,as a factor affecting the normal growth of A.fangsiao,egg protection has rarely been considered in previous behavioral studies.In this research,we analyzed the transcriptome profile of gene expression in A.fangsiao egg-unprotected larvae and egg-protected larvae,and identified 818 differentially expressed genes(DEGs).We used GO and KEGG enrichment analyses to search for metabolism-related DEGs.Protein-protein interaction networks were constructed to examine the interactions between metabolism-related genes.Twenty hub genes with multiple protein-protein interaction relationships or that were involved in multiple KEGG signaling pathways were obtained and verified by quantitative RT-PCR.We first studied the effects of egg protection on the metabolism of A.fangsiao larvae by means of protein-protein interaction networks,and the results provide va-luable gene resources for understanding the metabolism of invertebrate larvae.The data serve as a foundation for further research on the egg-protecting behavior of invertebrates.展开更多
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.31101891)the Hitech Research and Development Program of China(No.2012AA10A408)
文摘As an aquatic fish,the spotted halibut Verasper variegatus is highly susceptible to bacterial and virus infections.Tumor necrosis factor-alpha(TNF-α)as a cytokine could control the inflammatory responses.The functions of TNF-αin many species have been widely studied,particularly in mammals.However,little is known about the TNF-αfunctions in V.variegatus.We first cloned and sequenced the TNF-αgene in V.variegatus(VvTNF-α).The two conserved cysteine residues,transmembrane sequence,Thr-Leu motif,and TNF family signature,as well as the TA-rich motifs of its proteins related to inflammatory responses had high similarity to those of the other teleost and mammalian TNF-α.The phylogenetic analysis showed that VvTNF-αwas consistent with TNF-αgenes of other vertebrates.The VvTNF-αtranscripts were extensively distributed in the peripheral blood leukocytes(PBLs),spleen,and gill,indicating that the VvTNF-αhad a role in immune function.Furthermore,treatment with pathogen-associated molecular patterns(PAMPs)could induce a rapid and significant increase of VvTNF-αin the PBLs,which reveals that VvTNF-αdoes participate in the host immune responses against bacterial and viral pathogens.We found that VvTNF-αhad an interesting expression pattern during metamorphosis,showing that the flatfish TNF-αmay have some novel functions during specific developmental stages.In addition,the 3 D structure prediction of VvTNF-αprovided an indication of how it is likely to interact with other proteins.Therefore,VvTNF-αhas multiple functions,and provides valuable information to explore novel functions of TNF-α.
基金supported by the National High Technology R&D Program of China (2012AA10A402)the National Natural Science Foundation of China (31172385)
文摘Cold-inducible RNA-binding protein(CIRP) is a kind of RNA binding proteins that plays important roles in many physiological processes. The CIRP has been widely studied in mammals and amphibians since it was first cloned from mammals. On the contrary, there are little reports in teleosts. In this study, the Po CIRP gene of the Japanese flounder was cloned and sequenced. The genomic sequence consists of seven exons and six introns. The putative Po CIRP protein of flounder was 198 amino acid residues long containing the RNA recognition motif(RRM). Phylogenetic analysis showed that the flounder Po CIRP is highly conserved with other teleost CIRPs. The 5' flanking sequence was cloned by genome walking and many transcription factor binding sites were identified. There is a Cp Gs region located in promoter and exon I region and the methylation state is low. Quantitative real-time PCR analysis uncovered that Po CIRP gene was widely expressed in adult tissues with the highest expression level in the ovary. The m RNA of the Po CIRP was maternally deposited and the expression level of the gene was regulated up during the gastrula and neurula stages. In order to gain the information how the protein interacts with m RNA, we performed the modeling of the 3D structure of the flounder Po CIRP. The results showed a cleft existing the surface of the molecular. Taken together, the results indicate that the CIRP is a multifunctional molecular in teleosts and the findings about the structure provide valuable information for understanding the basis of this protein's function.
基金supported by the National Natural Sciences Foundation of China(No.31101891)the Hitech Research and Development Program of China(No.2012AA10A408)
文摘Interleukin-1 beta(IL-1β) is a cytokine involved in the control of inflammatory responses.The functions of IL-1β have been extensively studied in many vertebrates.However,few studies focused on the functions of IL-1β in the spotted halibut(Verasper variegatus),an endangered aquatic fish that easily suffers from bacterial and viral infections.We first cloned and sequenced the IL-1β gene from V.variegatus(Vv IL-1β).Its conservative interleukin-1 propeptide and interleukin-1/18 domains corresponding regions,characteristics of the IL-1 protein family and TA-rich motifs relating to inflammatory responses were similar to those of the IL-1β genes from other teleost and mammals.The phylogenetic analysis showed that Vv IL-1β was homologous to the IL-1β gene of other vertebrates.Treatment with pathogen-associated molecular patterns(PAMPs) induced a significant and rapid up-regulation of Vv IL-1β expression in the peripheral blood leukocytes(PBLs),revealing that Vv IL-1β is actively involved in the host immune responses against bacterial and viral pathogens.The expression level of Vv IL-1β was relatively high in the PBLs,gill,and intestine,indicating the immunological function of Vv IL-1β.Vv IL-1β also had interesting expression patterns at specific developmental stages,implying that the IL-1β of flatfishes may have some novel functions during embryonic development and metamorphosis.The 3 D structure prediction also provided information about how Vv IL-1β interacts with its receptor protein.These results suggested that Vv IL-1β may have multiple functions in V.variegatus and provided us valuable information for understanding the novel functions of IL-1β.
基金supported by the National Natural Science Foundation of China(No.31672646)the National Key R&D Program of China(No.2018YFD0900101)+1 种基金the Natural Science Foundation of Shandong Province(No.ZR2018BC052)the Science and Technology Project of Yantai University(No.2219008).
文摘GATA4,as a member of the GATA transcription factor family,plays a significant role in lineage specification and trans-differentiation of cells by regulating target gene expression.In this study,the potential transcription factor binding sites in the pro-moter of Paralichthys olivaceus sox2 gene were analyzed,and a GATA4-binding motif was identified.A cleaved amplified poly-morphic sequence-based binding assay experiment was performed,and the results showed a significant binding process between the Homo sapiens GATA4 recombinant protein and the DNA fragment of P.olivaceus sox2 promoter.Luciferase reporter assay revealed that GATA4 could improve the activity of the P.olivaceus sox2 promoter.Overexpression of P.olivaceus GATA4 could also increase the sox2 mRNA expression level in flounder brain cells.Overall,our results indicate that sox2 is a target gene of GATA4 in P.oliva-ceus.Sox2 is implicated in the maintenance of stem cell pluripotency and can reprogram differentiated cells to iPS cells.In addition,sox2 is involved in the development of the central nervous system(CNS).Therefore,this study may provide a reference for exploring the function of P.olivaceus GATA4 in the regulation of pluripotent stem cells and the development of the CNS.
基金supported by the earmarked fund for the Modern Agro-industry Technology Research System(No.CARS-49)the Natural Science Foundation of Shan-dong Province(No.ZR2019BC052)the National Natural Science Foundation of China(No.42006077).
文摘Marine organisms cannot grow and reproduce without proper metabolic regulation.Within a metabolic network,problems with a given link will affect the normal life activities of the organism.Many metabolic mechanisms associated with behaviors of Am-phioctopus fangsiao are still unclear.Moreover,as a factor affecting the normal growth of A.fangsiao,egg protection has rarely been considered in previous behavioral studies.In this research,we analyzed the transcriptome profile of gene expression in A.fangsiao egg-unprotected larvae and egg-protected larvae,and identified 818 differentially expressed genes(DEGs).We used GO and KEGG enrichment analyses to search for metabolism-related DEGs.Protein-protein interaction networks were constructed to examine the interactions between metabolism-related genes.Twenty hub genes with multiple protein-protein interaction relationships or that were involved in multiple KEGG signaling pathways were obtained and verified by quantitative RT-PCR.We first studied the effects of egg protection on the metabolism of A.fangsiao larvae by means of protein-protein interaction networks,and the results provide va-luable gene resources for understanding the metabolism of invertebrate larvae.The data serve as a foundation for further research on the egg-protecting behavior of invertebrates.