An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified and inserted into pET24a (+) vector. Then it expressed in E. c...An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified and inserted into pET24a (+) vector. Then it expressed in E. coli BL21 (DE3), and His-tag fusion protein of high yield was obtained. It was found that the fusion protein existed in E. coli mainly as inclusion bodies. The bioinformatics analysis indicates that LCN61 is C2H2 type zinc-finger protein containing four C2H2 zinc-finger motifs. This work provides a theory for functional research of lcn61 gene.展开更多
The Populus euphratica stress responsive zinc-finger protein gene PSTZ, which encodes a protein including typical Cys2/His2 zinc finger structure, was isolated by reverse transcription-polymerase chain reaction from P...The Populus euphratica stress responsive zinc-finger protein gene PSTZ, which encodes a protein including typical Cys2/His2 zinc finger structure, was isolated by reverse transcription-polymerase chain reaction from P. euphratica. Northern hybridization revealed that its expression was induced under drought and salt stress conditions. To examine its function, cDNA of the PSTZ gene, driven by the cauliflower mosaic virus 35S promoter, was cloned into a plant expression vector pBin438 and introduced into tobacco plants. Transgenic tobacco showed an enhanced salt tolerance, suggesting that PSTZ may play a role in plant responsiveness to salt stress.展开更多
The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses by eliminating viral mRNAs in the cytoplasm.In previous studies,we demonstrated that ZAP directly bi...The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses by eliminating viral mRNAs in the cytoplasm.In previous studies,we demonstrated that ZAP directly binds to the viral mRNAs and recruits the RNA exosome to degrade the target RNA.In this article,we provide evidence that a DEXH box RNA helicase,DHX30,is required for optimal antiviral activity of ZAP.Pull-down and co-immunoprecipitation assays demonstrated that DHX30 and ZAP interacted with each other via their N terminal domains.Downregulation of DHX30 with shRNAs reduced ZAP’s antiviral activity.These data implicate that DHX30 is a cellular factor involved in the antiviral function of ZAP.展开更多
The aim of the study was to investigate the correlation of zinc-finger protein 217 （ZNF217） gone ex- pression with the biological behavior of human ovarian cancer HO-8910 cells. Methods： The expression of ZNF217 in...The aim of the study was to investigate the correlation of zinc-finger protein 217 （ZNF217） gone ex- pression with the biological behavior of human ovarian cancer HO-8910 cells. Methods： The expression of ZNF217 in ovarian carcinoma cell line：s was detected by RT-PCR and Western blot, respectively. The biological behaviors of the transfectants were investigated by MTT, in vitro Boyden chamber and in vivo invasion assay, respectively. Results： RT-PCR and Western blotting revealed that transfection of ZNF217 into the HO-8910 cells significantly increased their proliferation along with mark- edly enhanced in vitro and in vivo invasion and metastatic abilities. MTT assay showed that the proliferation ability of pEGFP- N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells and HO-8910 cells （P 〈 0.001）. The Boyden chamber assay showed that the numbers of migrating pEGFP-N1-ZNF217/HO-8910, pEGFP-N1/HO-8910 and HO-8910 cells were （141.25 ± 13.91） cells/200 x field, （82.50 ± 11.73） cells/200 × field and （81.75 ± 12.12） cells/200 x field, respectively, with a significant difference between them （F = 29.274, P 〈 0.001）. The nude mouse experiment showed that the in vivo tumor formation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells （P 〈 0.001）. Conclusion： Based on these clinical and laboratory observations, we conclude that ZNF217 may contribute to ovarian cancer invasion and metastasis, and associated with worse clinical outcomes. We evaluated ZNF217＇s role as a biomarker of ovarian carcinogenesis and tumor progression in patient samples and explored possible molecular mechanisms in promoting tumor growth and invasion.展开更多
Background:LncRNA DLX6-AS1 has been uncovered to exert effects on various cancers.Nevertheless,the impacts of DLX6-AS1 on endometrial cancer(EC)development remained obscure.The study explored the influence of DLX6-AS1...Background:LncRNA DLX6-AS1 has been uncovered to exert effects on various cancers.Nevertheless,the impacts of DLX6-AS1 on endometrial cancer(EC)development remained obscure.The study explored the influence of DLX6-AS1 on EC progression via the microRNA(miR)-374a-3p/zinc-finger protein(ZFX)axis.Methods:EC cell lines were collected and DLX6-AS1,miR-374a-3p,and ZFX levels in EC cell lines were detected.The EC cells were transfected with DLX6-AS1,miR-374a-3p,and ZFX constructs to examine the biological functions of EC cells.The xenograft model was established for detecting tumor growth.Rescue experiments were conducted to verify the interaction of DLX6-AS1,miR-374a-3p,and ZFX in EC cells.Results:DLX6-AS1 and ZFX levels were elevated,while miR-374a-3p exhibited a reduced level in EC cells.Silencing DLX6-AS1 and elevated miR-374a-3p expressions repressed the biological activities of EC cells.Reduced DLX6-AS1 repressed tumor development.MiR-374a-3p silencing reversed the impacts of DLX6-AS1 silencing,while ZFX overexpression abrogated the impacts of miR-374a-3p elevation on EC cell growth.Mechanically,DLX6-AS1 was found to bind to miR-374a-3p,and miR-374a-3p targeted ZFX.Conclusion:DLX6-AS1 depletion restricts the malignant phenotype of EC cells.The study might provide novel therapeutic biomarkers for EC treatment.展开更多
The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses,including murine leukemia virus,Sindbis virus and Ebola virus,by targeting the viral mRNAs for degra...The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses,including murine leukemia virus,Sindbis virus and Ebola virus,by targeting the viral mRNAs for degradation.ZAP directly binds to the target viral mRNA and recruits the cellular RNA degradation machinery to degrade the RNA.No significant sequence similarity or obvious common motifs have been found in the so far identified target viral mRNAs.The minimum length of the target sequence is about 500 nt long.Short workable ZAP-binding RNAs should facilitate further studies on the ZAP-RNA interaction and characterization of such RNAs may provide some insights into the underlying mechanism.In this study,we used the SELEX method to isolate ZAP-binding RNA aptamers.After 21 rounds of selection,ZAP-binding aptamers were isolated.Sequence analysis revealed that they are G-rich RNAs with predicted stem-loop structures containing conserved“GGGUGG”and“GAGGG”motifs in the loop region.Insertion of the aptamer sequence into a luciferase reporter failed to render the reporter sensitive to ZAP.However,overexpression of the aptamers modestly but significantly reduced ZAP’s antiviral activity.Substitution of the conserved motifs of the aptamers significantly impaired their ZAP-binding ability and ZAP-antagonizing activity,suggesting that the RNA sequence is important for specific interaction between ZAP and the target RNA.The aptamers identified in this report should provide useful tools to further investigate the details of the interaction between ZAP and the target RNAs.展开更多
基金Supported by High Technology Research and Development Program of China (863 Program, No. 2006AA100309)
文摘An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified and inserted into pET24a (+) vector. Then it expressed in E. coli BL21 (DE3), and His-tag fusion protein of high yield was obtained. It was found that the fusion protein existed in E. coli mainly as inclusion bodies. The bioinformatics analysis indicates that LCN61 is C2H2 type zinc-finger protein containing four C2H2 zinc-finger motifs. This work provides a theory for functional research of lcn61 gene.
基金Supported by the National Key Technology Research and Development Program (2006BAD03A01)the Hi-Tech Research and Development Program of China (2007AA10Z106)the Key Program Project of Ministry of Education (104242).
文摘The Populus euphratica stress responsive zinc-finger protein gene PSTZ, which encodes a protein including typical Cys2/His2 zinc finger structure, was isolated by reverse transcription-polymerase chain reaction from P. euphratica. Northern hybridization revealed that its expression was induced under drought and salt stress conditions. To examine its function, cDNA of the PSTZ gene, driven by the cauliflower mosaic virus 35S promoter, was cloned into a plant expression vector pBin438 and introduced into tobacco plants. Transgenic tobacco showed an enhanced salt tolerance, suggesting that PSTZ may play a role in plant responsiveness to salt stress.
基金supported by the grant to Guangxia Gao from National Science Foundation of China(Grant No.81030030)by the grant toGuifang Chen from National Science Foundation(Grant No.30800053).
文摘The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses by eliminating viral mRNAs in the cytoplasm.In previous studies,we demonstrated that ZAP directly binds to the viral mRNAs and recruits the RNA exosome to degrade the target RNA.In this article,we provide evidence that a DEXH box RNA helicase,DHX30,is required for optimal antiviral activity of ZAP.Pull-down and co-immunoprecipitation assays demonstrated that DHX30 and ZAP interacted with each other via their N terminal domains.Downregulation of DHX30 with shRNAs reduced ZAP’s antiviral activity.These data implicate that DHX30 is a cellular factor involved in the antiviral function of ZAP.
基金Supported by grants from Medical Science and Technology Research Fund of Guangdong Province(No.WSTJJ20111110440104197405153780)the Dean Fund of Nanfang Hospital,Southern Medical University(No.2012B015)
文摘The aim of the study was to investigate the correlation of zinc-finger protein 217 （ZNF217） gone ex- pression with the biological behavior of human ovarian cancer HO-8910 cells. Methods： The expression of ZNF217 in ovarian carcinoma cell line：s was detected by RT-PCR and Western blot, respectively. The biological behaviors of the transfectants were investigated by MTT, in vitro Boyden chamber and in vivo invasion assay, respectively. Results： RT-PCR and Western blotting revealed that transfection of ZNF217 into the HO-8910 cells significantly increased their proliferation along with mark- edly enhanced in vitro and in vivo invasion and metastatic abilities. MTT assay showed that the proliferation ability of pEGFP- N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells and HO-8910 cells （P 〈 0.001）. The Boyden chamber assay showed that the numbers of migrating pEGFP-N1-ZNF217/HO-8910, pEGFP-N1/HO-8910 and HO-8910 cells were （141.25 ± 13.91） cells/200 x field, （82.50 ± 11.73） cells/200 × field and （81.75 ± 12.12） cells/200 x field, respectively, with a significant difference between them （F = 29.274, P 〈 0.001）. The nude mouse experiment showed that the in vivo tumor formation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells （P 〈 0.001）. Conclusion： Based on these clinical and laboratory observations, we conclude that ZNF217 may contribute to ovarian cancer invasion and metastasis, and associated with worse clinical outcomes. We evaluated ZNF217＇s role as a biomarker of ovarian carcinogenesis and tumor progression in patient samples and explored possible molecular mechanisms in promoting tumor growth and invasion.
基金supported by Shanghai Municipal Health Commission(Grant/Award No.20194Y0050).
文摘Background:LncRNA DLX6-AS1 has been uncovered to exert effects on various cancers.Nevertheless,the impacts of DLX6-AS1 on endometrial cancer(EC)development remained obscure.The study explored the influence of DLX6-AS1 on EC progression via the microRNA(miR)-374a-3p/zinc-finger protein(ZFX)axis.Methods:EC cell lines were collected and DLX6-AS1,miR-374a-3p,and ZFX levels in EC cell lines were detected.The EC cells were transfected with DLX6-AS1,miR-374a-3p,and ZFX constructs to examine the biological functions of EC cells.The xenograft model was established for detecting tumor growth.Rescue experiments were conducted to verify the interaction of DLX6-AS1,miR-374a-3p,and ZFX in EC cells.Results:DLX6-AS1 and ZFX levels were elevated,while miR-374a-3p exhibited a reduced level in EC cells.Silencing DLX6-AS1 and elevated miR-374a-3p expressions repressed the biological activities of EC cells.Reduced DLX6-AS1 repressed tumor development.MiR-374a-3p silencing reversed the impacts of DLX6-AS1 silencing,while ZFX overexpression abrogated the impacts of miR-374a-3p elevation on EC cell growth.Mechanically,DLX6-AS1 was found to bind to miR-374a-3p,and miR-374a-3p targeted ZFX.Conclusion:DLX6-AS1 depletion restricts the malignant phenotype of EC cells.The study might provide novel therapeutic biomarkers for EC treatment.
基金supported in part by Grants(to G.G.)from National Natural Science Foundation of China(Grant Nos.30470092 and 30530020)National Basic Research Program of China(973 Program)(Grant No.2006CB504302)of China.
文摘The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses,including murine leukemia virus,Sindbis virus and Ebola virus,by targeting the viral mRNAs for degradation.ZAP directly binds to the target viral mRNA and recruits the cellular RNA degradation machinery to degrade the RNA.No significant sequence similarity or obvious common motifs have been found in the so far identified target viral mRNAs.The minimum length of the target sequence is about 500 nt long.Short workable ZAP-binding RNAs should facilitate further studies on the ZAP-RNA interaction and characterization of such RNAs may provide some insights into the underlying mechanism.In this study,we used the SELEX method to isolate ZAP-binding RNA aptamers.After 21 rounds of selection,ZAP-binding aptamers were isolated.Sequence analysis revealed that they are G-rich RNAs with predicted stem-loop structures containing conserved“GGGUGG”and“GAGGG”motifs in the loop region.Insertion of the aptamer sequence into a luciferase reporter failed to render the reporter sensitive to ZAP.However,overexpression of the aptamers modestly but significantly reduced ZAP’s antiviral activity.Substitution of the conserved motifs of the aptamers significantly impaired their ZAP-binding ability and ZAP-antagonizing activity,suggesting that the RNA sequence is important for specific interaction between ZAP and the target RNA.The aptamers identified in this report should provide useful tools to further investigate the details of the interaction between ZAP and the target RNAs.
