ArticleProceedings of the National Academy of Sciences of the United States of America2001
High intranuclear mobility and dynamic clustering of the splicing factor U1 snRNP observed by single particle tracking.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 75 citations in OpenAlex.
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- Application of single molecule fluorescence microscopy to characterize the penetration of a large amphiphilic molecule in the stratum corneum of human skin.International journal of molecular sciences · 2015Article
- Mapping protein-specific micro-environments in live cells by fluorescence lifetime imaging of a hybrid genetic-chemical molecular rotor tag.Chemical communications (Cambridge, England) · 2012Article
- Protein-flexibility mediated coupling between photoswitching kinetics and surrounding viscosity of a photochromic fluorescent protein.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
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- Cell-penetrating HIV1 TAT peptides can generate pores in model membranes.Biophysical journal · 2010Article
- Assembly and mobility of exon-exon junction complexes in living cells.RNA (New York, N.Y.) · 2009Article
- Autonomy and robustness of translocation through the nuclear pore complex: a single-molecule study.The Journal of cell biology · 2008Article
- 4Pi microscopy of the nuclear pore complex.Biophysical journal · 2008Article
- Probing intranuclear environments at the single-molecule level.Biophysical journal · 2008Article
- Intranuclear binding kinetics and mobility of single native U1 snRNP particles in living cells.Molecular biology of the cell · 2006Article
- Enhancement of U4/U6 small nuclear ribonucleoprotein particle association in Cajal bodies predicted by mathematical modeling.Molecular biology of the cell · 2006Article
- Gene expression within a dynamic nuclear landscape.The EMBO journal · 2006Review
- Single synaptic vesicle tracking in individual hippocampal boutons at rest and during synaptic activity.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2005Article
- High-resolution near-field optical imaging of single nuclear pore complexes under physiological conditions.Biophysical journal · 2005Article
- Nuclear transport of single molecules: dwell times at the nuclear pore complex.The Journal of cell biology · 2005Article
- Using single-particle tracking to study nuclear trafficking of viral genes.Biophysical journal · 2004Article
- Three-dimensional tracking of single secretory granules in live PC12 cells.Biophysical journal · 2004Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Uridine-rich small nuclear ribonucleoproteins (U snRNPs) are components of the splicing machinery that removes introns from precursor mRNA. Like other splicing factors, U snRNPs are diffusely distributed throughout the nucleus and, in addition, are concentrated in distinct nuclear substructures referred to as speckles. We have examined the intranuclear distribution and mobility of the splicing factor U1 snRNP on a single-molecule level. Isolated U1 snRNPs were fluorescently labeled and incubated with digitonin-permeabilized 3T3 cells in the presence of Xenopus egg extract. By confocal microscopy, U1 snRNPs were found to be imported into nuclei, yielding a speckled intranuclear distribution. Employing a laser video-microscope optimized for high sensitivity and high speed, single U1 snRNPs were visualized and tracked at a spatial precision of 35 nm and a time resolution of 30 ms. The single-particle data revealed that U1 snRNPs occurred in small clusters that colocalized with speckles. In the clusters, U1 snRNPs resided for a mean decay time of 84 ms before leaving the optical slice in the direction of the optical axis, which corresponded to a mean effective diffusion coefficient of 1 microm(2)/s. An analysis of the trajectories of single U1 snRNPs revealed that at least three kinetic classes of low, medium, and high mobility were present. Moreover, the mean square displacements of these fractions were virtually independent of time, suggesting arrays of binding sites. The results substantiate the view that nuclear speckles are not rigid structures but highly dynamic domains characterized by a rapid turnover of U1 snRNPs and other splicing factors.
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