A Stochastic View of Spliceosome Assembly and Recycling in the Nucleus 英文参考文献.docVIP

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A Stochastic View of Spliceosome Assembly and Recycling in the Nucleus 英文参考文献.doc

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A Stochastic View of Spliceosome Assembly and Recycling in the Nucleus 英文参考文献

AStochasticViewofSpliceosomeAssembly andRecyclingintheNucleus Jose′ Rino1[,Teresa Carvalho1[,Jose′ Braga1[,Joana M.P.Desterro1,Reinhard Lu¨hrmann2,Maria Carmo-Fonseca1* 1InstitutodeMedicinaMolecular,UniversidadedeLisboa,Lisbon,Portugal,2DepartmentofCellularBiochemistry,Max-Planck-InstituteofBiophysicalChemistry,Go¨ttingen, Germany Howsplicingfactorsarerecruitedtonascenttranscriptsinthenucleusinordertoassemblespliceosomesonnewly synthesised pre-mRNAs is unknown. To address this question, we compared the intranuclear trafficking kinetics of small nuclear ribonucleoprotein particles (snRNP) and non-snRNP proteins in the presence and absence of splicing activity. Photobleaching experiments clearly show that spliceosomal proteins move continuously throughout the entire nucleus independently of ongoing transcription or splicing. Using quantitative experimental data, a mathematical model was applied for spliceosome assembly and recycling in the nucleus. The model assumes that splicing proteins move by Brownian diffusion and interact stochastically with binding sites located at different subnuclearcompartments.Inhibitionofsplicing,whichreducesthenumberofpre-mRNAbindingsitesavailablefor spliceosomeassembly,wasmodeledasadecreaseintheon-ratebindingconstantinthenucleoplasm.Simulationof microscopy experiments before and after splicing inhibition yielded results consistent with the experimental observations. Taken together, our data argue against the view that spliceosomal components are stored in nuclear speckles until a signal triggers their recruitment to nascent transcripts. Rather, the results suggest that splicing proteins are constantly diffusing throughout the entire nucleus and collide randomly and transiently with pre- mRNAs. Citation:RinoJ,CarvalhoT,BragaJ,DesterroJMP,Lu¨hrmannR,etal.(2007)Astochasticviewofspliceosomeassemblyandrecyclinginthenucleus.PLoSComputBiol3(10): e201.doi:10.1371/journal.pcbi.0030201 mediated transport is linked to the nuclear import of