Dynamics of Intracellular Clathrin/AP1-and Clathrin/AP3-Containing Carriers

• Tipo: Artigo em Revista Científica Internacional
• Data: 2012

Autores

Kural, C (Autor) Outra; Tacheva Grigorova, SK (Autor) Outra; Boulant, S (Autor) Outra; Cocucci, E (Autor) Outra; Baust, T (Autor) Outra; Duarte, D (Autor) FMUP; Kirchhausen, T (Autor) Outra

Revista

Título: Cell Reports

Vol. 2

Páginas: 1111-1119

ISSN: 2211-1247

Editora: Elsevier

Indexação

ISI Web of Knowledge - 45 Citações

Scopus - 45 Citações

Outras Informações

ID Authenticus: P-002-3GH

DOI: 10.1016/j.celrep.2012.09.025

Abstract (EN): 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, sigma adaptor subunits, or dynamin2) with a spatial precision of up to similar to 30 nm and a temporal resolution of similar to 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.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 9