水稻黑条矮缩病毒(RBSDV)S3和S10基因组非编码区对翻译的调控作用
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摘要
水稻黑条矮缩病毒(Rice black-streaked dwarf virus,RBSDV)基因组含有5'帽子,无3'poly(A)尾巴,且不同基因组片段的5'和3'末端序列均十分保守。本研究以RBSDV S3和S10为对象,分析其非编码区对翻译的调控作用。研究发现S3和S10的5'UTR在有无帽子时均可以正向调控报告基因Fluc的翻译,具备核糖体内部进入位点(IRES)活性,且5'末端的基因组保守序列及邻近序列与IRES活性密切相关;而对应的3'UTR则抑制5'UTR对翻译的正调控效应,其分子基础是5'UTR和3'UTR之间的RNA-RNA互作,且该互作也抑制帽子依赖型翻译的效率。这是首次关于有帽子植物RNA病毒中IRES元件的报道,研究结果对了解植物双链RNA病毒基因组非编码区对翻译的调控作用具有重要科学意义。
Rice black-streaked dwarf virus contains ten genome fragments with 5' cap and without 3'poly( A),and have conserved 5' and 3' terminal sequences among ten fragments. In this study,effect of the untranslated regions of S3 and S10 on translation was analyzed. 5'UTR of S3 or S10 can enhance the translation of FLuc reporter gene and possess IRES activity in the absence or presence of 5'cap structure. Core sequences of IRES from S3 or S10 were located at the 5'terminal conserved sequences and its adjacent sequences. However,corresponding 3'UTR inhibited the IRES activity of 5'UTR from S3 or S10,which is mediated by RNA-RNA interaction between 5'UTR and 3'UTR of S3 or S10. Moreover,this inhibition induced by potential RNA-RNA interaction between 5'UTR and 3'UTR also occurred in the presence of 5'cap. This is the first report of IRES in plant RNA viruses with 5'cap structure. In addition,this is the first report about effect of untranslated regions of dsRNA viruses on translation.
Rice black-streaked dwarf virus contains ten genome fragments with 5' cap and without 3'poly( A),and have conserved 5' and 3' terminal sequences among ten fragments. In this study,effect of the untranslated regions of S3 and S10 on translation was analyzed. 5'UTR of S3 or S10 can enhance the translation of FLuc reporter gene and possess IRES activity in the absence or presence of 5'cap structure. Core sequences of IRES from S3 or S10 were located at the 5'terminal conserved sequences and its adjacent sequences. However,corresponding 3'UTR inhibited the IRES activity of 5'UTR from S3 or S10,which is mediated by RNA-RNA interaction between 5'UTR and 3'UTR of S3 or S10. Moreover,this inhibition induced by potential RNA-RNA interaction between 5'UTR and 3'UTR also occurred in the presence of 5'cap. This is the first report of IRES in plant RNA viruses with 5'cap structure. In addition,this is the first report about effect of untranslated regions of dsRNA viruses on translation.
引文
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[12]Shikata E,Kitagawa Y.Rice black-streaked dwarf virus:its properties,morphology and intracellular localization[J].Virology,1997,77(2):826-842.
[13]Deng J,Dai L.Research progress on Rice black streaked dwarf virus(RBSDV)(in Chinese)[J].Chinese Agricultural Science Bulletin(中国农学通报),2012,28(18):209-212.
[14]Wang Z,Fang S,Xu J,et al.Sequence analysis of the complete genome of Rice black-streaked dwarf virus isolated from maize with rough dw arf disease[J].Virus Genes,2003,27(2):163-168.
[15]Wang D,Yu C,Liu S,et al.Structural alteration of a BYDV-like translation element(BTE)that attenuates p35 expression in three mild Tobacco bushy top virus isolates[J].Scientific Reports,2017,7(1):4213.
[16]Sweeney T R,Dhote V,Yu Y,et al.A distinct class of internal ribosomal entry site in members of the Kobuvirus and proposed Salivirus and Paraturdivirus genera of the Picornaviridae[J].Journal of Virology,2012,86(3):1468-1486.
[17]Lozano G,Martínez-Salas E.Structural insights into viral IRES-dependent translation mechanisms[J].Current Opinion in Virology,2015,12:113-120.
[2]Gallie D R.A tale of two termini:a functional interaction between the termini of an mRNA is a prerequisite for efficient translation initiation[J].Gene,1998,216(1):1-11.
[3]Sonenberg N,Hinnebusch A G.Regulation of translation initiation in eukaryotes:mechanisms and biological targets[J].Cell,2009,136(4):731-745.
[4]Kieft J S.Viral IRES RNA structures and ribosome interactions[J].Trends in Biochemical Sciences,2008,33(6):274-283.
[5]Jiang J,LalibertéJ F.The genome-linked protein VPg of plant viruses-a protein with many partners[J].Current Opinion in Virology,2011,1(5):347-354.
[6]Nicholson B L,White K A.3’Cap-independent translation enhancers of positive-strand RNA plant viruses[J].Current Opinion in Virology,2011,1(5):373-380.
[7]Simon A E,Miller W A.3'cap-independent translation enhancers of plant viruses[J].Annual Review of Microbiology,2013,67:21-42.
[8]Zhang J,Roberts R,Rakotondrafara A M.The role of the 5'untranslated regions of Potyviridae in translation[J].Virus Research,2015,206:74-81.
[9]Wang D,Yuan X.Research advances in cap-independent translation in plant RNAviruses(in Chinese)[J].Chinese Journal of Virology(病毒学报),2017,33(4):652-660.
[10]Eskelin K,HafrénA,Rantalainen K I,et al.Potyviral VPg enhances viral RNA translation and inhibits reporter mRNA translation in planta[J].Journal of Virology,2011,85(17):9210-9221.
[11]Weingarten-Gabbay S,Elias-Kirma S,Nir R,et al.Systematic discovery of cap-independent translation sequences in human and viral genomes[J].Science,2016,351(6270),aad4939.
[12]Shikata E,Kitagawa Y.Rice black-streaked dwarf virus:its properties,morphology and intracellular localization[J].Virology,1997,77(2):826-842.
[13]Deng J,Dai L.Research progress on Rice black streaked dwarf virus(RBSDV)(in Chinese)[J].Chinese Agricultural Science Bulletin(中国农学通报),2012,28(18):209-212.
[14]Wang Z,Fang S,Xu J,et al.Sequence analysis of the complete genome of Rice black-streaked dwarf virus isolated from maize with rough dw arf disease[J].Virus Genes,2003,27(2):163-168.
[15]Wang D,Yu C,Liu S,et al.Structural alteration of a BYDV-like translation element(BTE)that attenuates p35 expression in three mild Tobacco bushy top virus isolates[J].Scientific Reports,2017,7(1):4213.
[16]Sweeney T R,Dhote V,Yu Y,et al.A distinct class of internal ribosomal entry site in members of the Kobuvirus and proposed Salivirus and Paraturdivirus genera of the Picornaviridae[J].Journal of Virology,2012,86(3):1468-1486.
[17]Lozano G,Martínez-Salas E.Structural insights into viral IRES-dependent translation mechanisms[J].Current Opinion in Virology,2015,12:113-120.