Dynamics of intracellular clathrin/AP1- and clathrin/AP3-containing carriers
Summary Clathrin/AP1- and clathrin/AP3-coated vesicular carriers originate from endosomes and the trans-Golgi network. Here, we report the real-time visualization of these structures in living cells reliably tracked by rapid, three-dimensional imaging with the use of a spinning-disk confocal microsc...
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| Hauptverfasser: | , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
29 November 2012
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| In: |
Cell reports
Year: 2012, Jahrgang: 2, Heft: 5, Pages: 1111-1119 |
| ISSN: | 2211-1247 |
| DOI: | 10.1016/j.celrep.2012.09.025 |
| Online-Zugang: | Verlag, Volltext: http://dx.doi.org/10.1016/j.celrep.2012.09.025 Verlag, Volltext: http://www.sciencedirect.com/science/article/pii/S2211124712003294 
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| Verfasserangaben: | Comert Kural, Silvia K. Tacheva-Grigorova, Steeve Boulant, Emanuele Cocucci, Thorsten Baust, Delfim Duarte, and Tom Kirchhausen |
| Zusammenfassung: | Summary Clathrin/AP1- and clathrin/AP3-coated vesicular carriers originate from endosomes and the trans-Golgi network. Here, we report the real-time visualization of these structures in living cells reliably tracked by rapid, three-dimensional imaging with the use of a spinning-disk confocal microscope. We imaged relatively sparse, diffraction-limited, fluorescent objects containing chimeric fluorescent protein (clathrin light chain, σ adaptor subunits, or dynamin2) with a spatial precision of up to ∼30 nm and a temporal resolution of ∼1 s. The dynamic characteristics of the intracellular clathrin/AP1 and clathrin/AP3 carriers are similar to those of endocytic clathrin/AP2 pits and vesicles; the clathrin/AP1 coats are, on average, slightly shorter-lived than their AP2 and AP3 counterparts. We confirmed that although dynamin2 is recruited as a burst to clathrin/AP2 pits immediately before their budding from the plasma membrane, we found no evidence supporting a similar association of dynamin2 with clathrin/AP1 or clathrin/AP3 carriers at any stage during their lifetime. We found no effects of chemical inhibitors of dynamin function or the K44A dominant-negative mutant of dynamin on AP1 and AP3 dynamics. This observation suggests that an alternative budding mechanism, yet to be discovered, is responsible for the scission step of clathrin/AP1 and clathrin/AP3 carriers. |
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| Beschreibung: | Gesehen am 06.11.2018 |
| Beschreibung: | Online Resource |
| ISSN: | 2211-1247 |
| DOI: | 10.1016/j.celrep.2012.09.025 |