mRNA单分子动态成像技术研究进展
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摘要
转录后成熟的mRNA与RNA结合蛋白结合,形成信使核糖核蛋白复合物,之后在胞质内沿细胞骨架转运并定位到亚细胞特定区域。在此过程中,mRNA的转运及调控对于mRNA的功能、蛋白质的定位和细胞极性生长的维持发挥着重要的作用。近年来,活细胞单分子成像技术的发展,为人们深入了解mRNA的动态调控提供了新的契机。通过对活细胞中mRNA和RNA结合蛋白的标记,我们不仅可以检测mRNA的动力学特征,还可以原位检测新合成的蛋白质。本文主要介绍mRNA的定位和翻译过程,重点叙述单分子动态成像技术在分析mRNA动态调控中的应用。活细胞单分子成像技术可以揭示mRNA翻译的动力学特性,并促进我们对mRNA命运的理解,为后续研究mRNA的动态调控提供参考。
Post-transcriptional mature mRNA binds to RNA-binding proteins to form messenger ribonucleoprotein complexes,which are then transported along the cytoskeleton and localized in specific subcellular regions. In this process,mRNA transport and regulation play an important role in the function of mRNA,protein localization and maintenance of cell polarity. In recent years,the development of single-molecule imaging of living cells has provided a new opportunity to understand the dynamic regulation of mRNA. By labeling mRNA and RNA binding proteins in living cells,we can detect not only the dynamic characteristics of mRNA,but also the newly synthesized proteins in situ. This article mainly introduces the localization and translation of mRNA,and focuses on the application of single-molecule dynamic imaging in the analysis of dynamic regulation of mRNA. Living cell single-molecule imaging can reveal the dynamics of mRNA translation,promote our understanding of the fate of mRNA,and provide a reference for the follow-up study of the dynamic regulation of mRNA.
Post-transcriptional mature mRNA binds to RNA-binding proteins to form messenger ribonucleoprotein complexes,which are then transported along the cytoskeleton and localized in specific subcellular regions. In this process,mRNA transport and regulation play an important role in the function of mRNA,protein localization and maintenance of cell polarity. In recent years,the development of single-molecule imaging of living cells has provided a new opportunity to understand the dynamic regulation of mRNA. By labeling mRNA and RNA binding proteins in living cells,we can detect not only the dynamic characteristics of mRNA,but also the newly synthesized proteins in situ. This article mainly introduces the localization and translation of mRNA,and focuses on the application of single-molecule dynamic imaging in the analysis of dynamic regulation of mRNA. Living cell single-molecule imaging can reveal the dynamics of mRNA translation,promote our understanding of the fate of mRNA,and provide a reference for the follow-up study of the dynamic regulation of mRNA.
引文
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[22]ARAVA Y,WANG Y,STOREY J D,et al.Genomewide analysis of mRNA translation profiles in Saccharomyces cerevisiae[J].Proc Natl Acad Sci U SA,2003,100(7):3889-3894.
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[25]INGOLIA N T,GHAEMMAGHAMI S,NEMMAN J R,et al.Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling[J].Science,2009,324(5924):218-223
[26]HERAUD-FARLOW JE,SHARANGDHAR T,LI X,et al.Staufen2 regulates neuronal target RNAs[J].Cell Rep,2013,5(6):1511-1518.
[27]KWON S.Single-molecule fluorescence in situ hybridization:quantitative imaging of single RNAmolecules[J].BMB Rep,2013,46(2):65-72.
[28]PATEL V L,MITRA S,HARRIS R et al.Spatial arrangement of an RNA zipcode identifies mRNAs under post-transcriptional control[J].Genes Dev,2012,26(1):43-53.
[29]NICO BATTICH,THOMAS STOEGER,LUCASPELKMANS.Image-based transcriptomics in thousands of single human cells at single-molecule resolution[J].Nat Methods,2013,10(11):1127-1133.
[30]HALSTEAD J M,LIONNET T,WILBERTZ J H,et al.An RNA biosensor for imaging the first round of translation from single cells to living animals[J].Science,2015,347(6228):1367-1671.
[31]QI L S,LARSON M H,GILBERT L A,et al.Repurposing CRISPR as an RNA-guided patform for sequence-specific cotrol of gene expressioon[J].Cell,2013,152(5):1173-1183.
[32]BERTRAND E,CHARTRAND P,SCHAEFER M,et al.Localization of ASH1 mRNA particles in living yeast[J].Mol Cell,1998,2(4):437-445.
[33]SCHNEIDER D,TUERK C,GOLD L.Selection of high affinity RNA ligands to the bacteriophage R17 coat protein[J].J Mol Biol,1992,228(3):862-869.
[34]WU B,MISKOLCI V,SATO H,et al.Synonymous modification results in high-fidelity gene expression of repetitive protein and nucleotide sequences[J].Genes Dev,2015,29(8):876-886.
[35]WU B,CHAO J A,SINGER R H.Fluorescence fluctuation spectroscopy enables quantitative imaging of single mRNAs in living cells[J].Biophys J,2012,102(12):2936-2944.
[36]WELLS A L,CONDEELIS J S,SINGER R H,et al.Imaging real-time gene expression in Mammalian cells with single-transcript resolution[J].CSH Protoc,2007,2007(11):pdb.prot4869.
[37]TANENBAUM M E,GILBERT L A,QI L S,et al.Aprotein-tagging system for signal amplification in gene expression and fluorescence imaging[J].Cell,2014,159(3):635-646.
[38]WU BIN,ELISCOVICH CAROLINA,YOUNG J,et al.Translation dynamics of single mRNAs in live cells and neurons[J].Science,2016,352(6292):1430-1435.
[39]YAN X,HOEK TIM A,VALE RONALD D,et al.Dynamics of translation of single mRNA molecules in vivo[J].Cell,2016,165(4):976-989.
[40]WANG CHONG,HAN BORAN,ZHOU RUO BO,et al.Real-time imaging of translation on single mRNAtranscripts in live cells[J].Cell,2016,165(4):990-1001.
[2]JUNG H,GKOGKAS C G,SONENBERG N,et al.Remote control of gene function by local translation[J].Cell,2014,157(1):26-40.
[3]XING L,BASSELL G J.mRNA localization:an orchestration of assembly,traffic and synthesis[J].Traffic,2013,14(1):2-14.
[4]石磊.mRNA定位的主动运输机制[J].国外医学分子生物学分册,2002,24(6):344-348.
[5]GLISOVIC T,BACHORIK J L,YONG J,et al.RNA-binding proteins and post-transcriptional gene regulation[J].FEBS Lett,2008,582(14):1977-1986.
[6]宋西娇,廖振凤,谢礼,等.免疫胶体金标记法研究筛管特异质体中微丝来源的细胞骨架成分[J].电子显微学报,2018,37(4):376-380.
[7]CHUDINOVA E M,NADEZHDINA E S.Interactions between the translation machinery and microtubules[J].Biochemistry,2018,83(1):S176-S189.
[8]HIROKAWA N,TANAKA Y.Kinesin superfamily proteins(KIFs):various functions and their relevance for important phenomena in life and diseases[J].Exp Cell Res,2015,334(1):16-25.
[9]TAKANO K,MIKI T,KATAHIRA J,et al.NXF2 is involved in cytoplasmic mRNA dynamics through interactions with motor proteins[J].Nucleic Acids Res,2007,35(8):2513-2521.
[10]LIU Y,SALTER H K,HOLDING A N,et al.Bicaudal-D uses a parallel:homodimeric coiled coil with heterotypic registry to coordinate recruitment of cargos to dynein[J].Genes Dev,2013,27(11):1233-1246.
[11]CALLIARI A,FARIAS J,PUPPO A,et al.Myosin Va associates with mRNA in ribonucleoprotein particles present in myelinated peripheral axons and in the central nervous system[J].Dev Neurobiol,2014,74(3):382-396.
[12]郑超星,马小凤,张永华,等.真核生物mRNA翻译起始机制的研究进展[J].遗传,2018,40(8):607-619.
[13]LACERDA R,MENEZES J,ROMAO L.More than just scanning:the importance of cap-independent mRNAtranslation initiation for cellular stress response and cancer[J].Cell Mol Life Sci,2017,74(9):1659-1680.
[14]余光创,秦宜德,伯晓晨,等.依赖于5'端非编码区高级结构的真核生物mRNA翻译调控[J].中国生物化学与分子生物学报,2007,23(11):881-887.
[15]潘昱婷,徐麒麟,康波.真核生物mRNA非翻译区结构特征与mRNA翻译[J].中国生物化学与分子生物学报,2016,32(9):990-993.
[16]GUO S,LU H.Conjunction of G-quadruplex and stemloop in the 5'untranslated region of mouse hepatocyte nuclear factor 4-alpha1 mediates strong inhibition of protein expression[J].Mol Cell Biochem,2018,446(1/2):73-81.
[17]郝亚静,骆健俊,张宝,等.RNA翻译的复杂性:不翻译、部分翻译、从头翻译及过度翻译[J].生物化学与生物物理进展,2017,44(7):547-556.
[18]谢兆辉,曾强成,沈亮,等.真核生物翻译过程中的mRNA质量控制[J].生物化学与生物物理进展,2013,40(1):22-29
[19]AAKALU G,SMITH W B,NGUYEN N,et al.Dynamic visualization of local protein synthesis in hippocampal neurons[J].Neuron,2001,30(2):489-502.
[20]LEUNG K M,van HORCK F P,LIN A C,et al.Asymmetrical beta-actin mRNA translation in growth cones mediates attractive turning to netrin-1[J].Nat Neurosci,2006,9(10):1247-1256.
[21]YU J,XIAO J,REN X,et al.Probing gene expression in live cells,one protein molecule at a time[J].Science,2006,311(5767):1600-1603.
[22]ARAVA Y,WANG Y,STOREY J D,et al.Genomewide analysis of mRNA translation profiles in Saccharomyces cerevisiae[J].Proc Natl Acad Sci U SA,2003,100(7):3889-3894.
[23]BRAR G A,WEISSMAN J S.Ribosome profiling reveals the what,when,where and how of protein synthesis[J].Nat Rev Mol Cell Bio,2015,16(11):651.
[24]INGOLIA N T.Ribosome profiling:new views of translation,from single codons to genome scale[J].Nat Rev Genet,2014,15(3):205-213.
[25]INGOLIA N T,GHAEMMAGHAMI S,NEMMAN J R,et al.Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling[J].Science,2009,324(5924):218-223
[26]HERAUD-FARLOW JE,SHARANGDHAR T,LI X,et al.Staufen2 regulates neuronal target RNAs[J].Cell Rep,2013,5(6):1511-1518.
[27]KWON S.Single-molecule fluorescence in situ hybridization:quantitative imaging of single RNAmolecules[J].BMB Rep,2013,46(2):65-72.
[28]PATEL V L,MITRA S,HARRIS R et al.Spatial arrangement of an RNA zipcode identifies mRNAs under post-transcriptional control[J].Genes Dev,2012,26(1):43-53.
[29]NICO BATTICH,THOMAS STOEGER,LUCASPELKMANS.Image-based transcriptomics in thousands of single human cells at single-molecule resolution[J].Nat Methods,2013,10(11):1127-1133.
[30]HALSTEAD J M,LIONNET T,WILBERTZ J H,et al.An RNA biosensor for imaging the first round of translation from single cells to living animals[J].Science,2015,347(6228):1367-1671.
[31]QI L S,LARSON M H,GILBERT L A,et al.Repurposing CRISPR as an RNA-guided patform for sequence-specific cotrol of gene expressioon[J].Cell,2013,152(5):1173-1183.
[32]BERTRAND E,CHARTRAND P,SCHAEFER M,et al.Localization of ASH1 mRNA particles in living yeast[J].Mol Cell,1998,2(4):437-445.
[33]SCHNEIDER D,TUERK C,GOLD L.Selection of high affinity RNA ligands to the bacteriophage R17 coat protein[J].J Mol Biol,1992,228(3):862-869.
[34]WU B,MISKOLCI V,SATO H,et al.Synonymous modification results in high-fidelity gene expression of repetitive protein and nucleotide sequences[J].Genes Dev,2015,29(8):876-886.
[35]WU B,CHAO J A,SINGER R H.Fluorescence fluctuation spectroscopy enables quantitative imaging of single mRNAs in living cells[J].Biophys J,2012,102(12):2936-2944.
[36]WELLS A L,CONDEELIS J S,SINGER R H,et al.Imaging real-time gene expression in Mammalian cells with single-transcript resolution[J].CSH Protoc,2007,2007(11):pdb.prot4869.
[37]TANENBAUM M E,GILBERT L A,QI L S,et al.Aprotein-tagging system for signal amplification in gene expression and fluorescence imaging[J].Cell,2014,159(3):635-646.
[38]WU BIN,ELISCOVICH CAROLINA,YOUNG J,et al.Translation dynamics of single mRNAs in live cells and neurons[J].Science,2016,352(6292):1430-1435.
[39]YAN X,HOEK TIM A,VALE RONALD D,et al.Dynamics of translation of single mRNA molecules in vivo[J].Cell,2016,165(4):976-989.
[40]WANG CHONG,HAN BORAN,ZHOU RUO BO,et al.Real-time imaging of translation on single mRNAtranscripts in live cells[J].Cell,2016,165(4):990-1001.