The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells

被引：81

作者：

Huranova, Martina ^{[1
]}

Ivani, Ivan ^{[1
]}

Benda, Ales ^{[2
]}

Poser, Ina ^{[3
]}

Brody, Yehuda ^{[4
,5
]}

Hof, Martin ^{[2
]}

Shav-Tal, Yaron ^{[4
,5
]}

Neugebauer, Karla M. ^{[3
]}

Stanek, David ^{[1
]}

机构：

[1] Acad Sci Czech Republic, Inst Mol Genet, CR-14220 Prague, Czech Republic

[2] Acad Sci Czech Republic, J Heyrovsky Inst Phys Chem, Prague 14220, Czech Republic

[3] Max Planck Inst Mol Cell Biol & Genet, D-01307 Dresden, Germany

[4] Bar Ilan Univ, Mina & Everard Goodman Fac Life Sci, IL-52900 Ramat Gan, Israel

[5] Bar Ilan Univ, Inst Nanotechnol & Adv Mat, IL-52900 Ramat Gan, Israel

来源：

JOURNAL OF CELL BIOLOGY | 2010年 / 191卷 / 01期

关键词：

FLUORESCENCE CORRELATION SPECTROSCOPY; IN-SITU TRANSCRIPTION; ASSEMBLY DYNAMICS; MAMMALIAN-CELLS; A-COMPLEXES; PENTA-SNRNP; U1; SNRNP; VIVO; SPLICEOSOME; BINDING;

D O I：

10.1083/jcb.201004030

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Precursor messenger RNA (pre-mRNA) splicing is catalyzed by the spliceosome, a large ribonucleoprotein (RNP) complex composed of five small nuclear RNP particles (snRNPs) and additional proteins. Using live cell imaging of GFP-tagged snRNP components expressed at endogenous levels, we examined how the spliceosome assembles in vivo. A comprehensive analysis of snRNP dynamics in the cell nucleus enabled us to determine snRNP diffusion throughout the nucleoplasm as well as the interaction rates of individual snRNPs with pre-mRNA. Core components of the spliceosome, U2 and U5 snRNPs, associated with pre-mRNA for 15-30 s, indicating that splicing is accomplished within this time period. Additionally, binding of U1 and U4/U6 snRNPs with pre-mRNA occurred within seconds, indicating that the interaction of individual snRNPs with pre-mRNA is distinct. These results are consistent with the predictions of the step-wise model of spliceosome assembly and provide an estimate on the rate of splicing in human cells.

引用

页码：75 / 86

页数：12

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