Ben-Ari, Y., Brody, Y., Kinor, N., Mor, A., Tsukamoto, T., Spector, D. L., Singer, R. H., Shav-Tal, Y. (May 2010) The life of an mRNA in space and time. Journal of Cell Science, 123 (10). pp. 1761-1774. ISSN 0021-9533
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Abstract
Nuclear transcribed genes produce mRNA transcripts destined to travel from the site of transcription to the cytoplasm for protein translation. Certain transcripts can be further localized to specific cytoplasmic regions. We examined the life cycle of a transcribed beta-actin mRNA throughout gene expression and localization, in a cell system that allows the in vivo detection of the gene locus, the transcribed mRNAs and the cytoplasmic beta-actin protein that integrates into the actin cytoskeleton. Quantification showed that RNA polymerase II elongation progressed at a rate of 3.3 kb/minute and that transactivator binding to the promoter was transient (40 seconds), and demonstrated the unique spatial structure of the coding and non-coding regions of the integrated gene within the transcription site. The rates of gene induction were measured during interphase and after mitosis, demonstrating that daughter cells were not synchronized in respect to transcription initiation of the studied gene. Comparison of the spatial and temporal kinetics of nucleoplasmic and cytoplasmic mRNA transport showed that the beta-actin-localization response initiates from the existing cytoplasmic mRNA pool and not from the newly synthesized transcripts arising after gene induction. It was also demonstrated that mechanisms of random movement were predominant in mediating the efficient translocation of mRNA in the eukaryotic cell.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | mRNA dynamics Gene expression Transcription Live cell imaging GENE-EXPRESSION TRANSCRIPTION FACTORIES POLYMERASE-II LIVING CELLS REAL-TIME IN-VIVO INTRACELLULAR-LOCALIZATION FLUORESCENCE RECOVERY BINDING-PROTEINS DYNAMICS |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > miRNA bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > miRNA |
| CSHL Authors: | |
| Communities: | CSHL labs > Spector lab CSHL Cancer Center Shared Resources > DNA Sequencing Service CSHL Cancer Center Shared Resources > Microscopy Service |
| Depositing User: | CSHL Librarian |
| Date: | May 2010 |
| Date Deposited: | 26 Sep 2011 13:54 |
| Last Modified: | 26 Jan 2015 15:49 |
| PMCID: | PMC2864715 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/15352 |
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