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Spatial mapping of splicing factor complexes involved in exon and intron definition
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| Zeitschriftentitel: | The Journal of Cell Biology |
|---|---|
| Personen und Körperschaften: | , , , , |
| In: | The Journal of Cell Biology, 181, 2008, 6, S. 921-934 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Rockefeller University Press
|
| Schlagwörter: |
| author_facet |
Ellis, Jonathan D. Llères, David Denegri, Marco Lamond, Angus I. Cáceres, Javier F. Ellis, Jonathan D. Llères, David Denegri, Marco Lamond, Angus I. Cáceres, Javier F. |
|---|---|
| author |
Ellis, Jonathan D. Llères, David Denegri, Marco Lamond, Angus I. Cáceres, Javier F. |
| spellingShingle |
Ellis, Jonathan D. Llères, David Denegri, Marco Lamond, Angus I. Cáceres, Javier F. The Journal of Cell Biology Spatial mapping of splicing factor complexes involved in exon and intron definition Cell Biology |
| author_sort |
ellis, jonathan d. |
| spelling |
Ellis, Jonathan D. Llères, David Denegri, Marco Lamond, Angus I. Cáceres, Javier F. 1540-8140 0021-9525 Rockefeller University Press Cell Biology http://dx.doi.org/10.1083/jcb.200710051 <jats:p>We have analyzed the interaction between serine/arginine-rich (SR) proteins and splicing components that recognize either the 5′ or 3′ splice site. Previously, these interactions have been extensively characterized biochemically and are critical for both intron and exon definition. We use fluorescence resonance energy transfer (FRET) microscopy to identify interactions of individual SR proteins with the U1 small nuclear ribonucleoprotein (snRNP)–associated 70-kD protein (U1 70K) and with the small subunit of the U2 snRNP auxiliary factor (U2AF35) in live-cell nuclei. We find that these interactions occur in the presence of RNA polymerase II inhibitors, demonstrating that they are not exclusively cotranscriptional. Using FRET imaging by means of fluorescence lifetime imaging microscopy (FLIM), we map these interactions to specific sites in the nucleus. The FLIM data also reveal a previously unknown interaction between HCC1, a factor related to U2AF65, with both subunits of U2AF. Spatial mapping using FLIM-FRET reveals differences in splicing factors interactions within complexes located in separate subnuclear domains.</jats:p> Spatial mapping of splicing factor complexes involved in exon and intron definition The Journal of Cell Biology |
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The Journal of Cell Biology |
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| title |
Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_unstemmed |
Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_full |
Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_fullStr |
Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_full_unstemmed |
Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_short |
Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_sort |
spatial mapping of splicing factor complexes involved in exon and intron definition |
| topic |
Cell Biology |
| url |
http://dx.doi.org/10.1083/jcb.200710051 |
| publishDate |
2008 |
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921-934 |
| description |
<jats:p>We have analyzed the interaction between serine/arginine-rich (SR) proteins and splicing components that recognize either the 5′ or 3′ splice site. Previously, these interactions have been extensively characterized biochemically and are critical for both intron and exon definition. We use fluorescence resonance energy transfer (FRET) microscopy to identify interactions of individual SR proteins with the U1 small nuclear ribonucleoprotein (snRNP)–associated 70-kD protein (U1 70K) and with the small subunit of the U2 snRNP auxiliary factor (U2AF35) in live-cell nuclei. We find that these interactions occur in the presence of RNA polymerase II inhibitors, demonstrating that they are not exclusively cotranscriptional. Using FRET imaging by means of fluorescence lifetime imaging microscopy (FLIM), we map these interactions to specific sites in the nucleus. The FLIM data also reveal a previously unknown interaction between HCC1, a factor related to U2AF65, with both subunits of U2AF. Spatial mapping using FLIM-FRET reveals differences in splicing factors interactions within complexes located in separate subnuclear domains.</jats:p> |
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| author | Ellis, Jonathan D., Llères, David, Denegri, Marco, Lamond, Angus I., Cáceres, Javier F. |
| author_facet | Ellis, Jonathan D., Llères, David, Denegri, Marco, Lamond, Angus I., Cáceres, Javier F., Ellis, Jonathan D., Llères, David, Denegri, Marco, Lamond, Angus I., Cáceres, Javier F. |
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| description | <jats:p>We have analyzed the interaction between serine/arginine-rich (SR) proteins and splicing components that recognize either the 5′ or 3′ splice site. Previously, these interactions have been extensively characterized biochemically and are critical for both intron and exon definition. We use fluorescence resonance energy transfer (FRET) microscopy to identify interactions of individual SR proteins with the U1 small nuclear ribonucleoprotein (snRNP)–associated 70-kD protein (U1 70K) and with the small subunit of the U2 snRNP auxiliary factor (U2AF35) in live-cell nuclei. We find that these interactions occur in the presence of RNA polymerase II inhibitors, demonstrating that they are not exclusively cotranscriptional. Using FRET imaging by means of fluorescence lifetime imaging microscopy (FLIM), we map these interactions to specific sites in the nucleus. The FLIM data also reveal a previously unknown interaction between HCC1, a factor related to U2AF65, with both subunits of U2AF. Spatial mapping using FLIM-FRET reveals differences in splicing factors interactions within complexes located in separate subnuclear domains.</jats:p> |
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| spelling | Ellis, Jonathan D. Llères, David Denegri, Marco Lamond, Angus I. Cáceres, Javier F. 1540-8140 0021-9525 Rockefeller University Press Cell Biology http://dx.doi.org/10.1083/jcb.200710051 <jats:p>We have analyzed the interaction between serine/arginine-rich (SR) proteins and splicing components that recognize either the 5′ or 3′ splice site. Previously, these interactions have been extensively characterized biochemically and are critical for both intron and exon definition. We use fluorescence resonance energy transfer (FRET) microscopy to identify interactions of individual SR proteins with the U1 small nuclear ribonucleoprotein (snRNP)–associated 70-kD protein (U1 70K) and with the small subunit of the U2 snRNP auxiliary factor (U2AF35) in live-cell nuclei. We find that these interactions occur in the presence of RNA polymerase II inhibitors, demonstrating that they are not exclusively cotranscriptional. Using FRET imaging by means of fluorescence lifetime imaging microscopy (FLIM), we map these interactions to specific sites in the nucleus. The FLIM data also reveal a previously unknown interaction between HCC1, a factor related to U2AF65, with both subunits of U2AF. Spatial mapping using FLIM-FRET reveals differences in splicing factors interactions within complexes located in separate subnuclear domains.</jats:p> Spatial mapping of splicing factor complexes involved in exon and intron definition The Journal of Cell Biology |
| spellingShingle | Ellis, Jonathan D., Llères, David, Denegri, Marco, Lamond, Angus I., Cáceres, Javier F., The Journal of Cell Biology, Spatial mapping of splicing factor complexes involved in exon and intron definition, Cell Biology |
| title | Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_full | Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_fullStr | Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_full_unstemmed | Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_short | Spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_sort | spatial mapping of splicing factor complexes involved in exon and intron definition |
| title_unstemmed | Spatial mapping of splicing factor complexes involved in exon and intron definition |
| topic | Cell Biology |
| url | http://dx.doi.org/10.1083/jcb.200710051 |