The AP‐2 complex is required for proper temporal and spatial dynamics of endocytic patches in fission yeast

Summary In metazoans the AP‐2 complex has a well‐defined role in clathrin‐mediated endocytosis. By contrast, its direct role in endocytosis in unicellular eukaryotes has been questioned. Here, we report co‐ immunoprecipitation between the fission yeast AP‐2 component Apl3p and clathrin, as well as t...

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Published in:Molecular microbiology 2016-05, Vol.100 (3), p.409-424
Main Authors: de León, Nagore, Hoya, Marta, Curto, M.‐Angeles, Moro, Sandra, Yanguas, Francisco, Doncel, Cristina, Valdivieso, M.‐Henar
Format: Article
Language:English
Subjects:
Adaptor Protein Complex 2 - genetics
Cell Wall - metabolism
Clathrin Light Chains - genetics
Clathrin Light Chains - metabolism
Endocytosis
Endocytosis - genetics
Endocytosis - physiology
Extracellular Matrix - metabolism
Metazoa
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Schizosaccharomyces - genetics
Schizosaccharomyces - metabolism
Schizosaccharomyces pombe
Schizosaccharomyces pombe Proteins - genetics
Schizosaccharomyces pombe Proteins - metabolism
Yeast
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Summary:Summary In metazoans the AP‐2 complex has a well‐defined role in clathrin‐mediated endocytosis. By contrast, its direct role in endocytosis in unicellular eukaryotes has been questioned. Here, we report co‐ immunoprecipitation between the fission yeast AP‐2 component Apl3p and clathrin, as well as the genetic interactions between apl3Δ and clc1 and sla2Δ/end4Δ mutants. Furthermore, a double clc1 apl3Δ mutant was found to be defective in FM4‐64 uptake. In an otherwise wild‐type strain, apl3Δ cells exhibit altered dynamics of the endocytic sites, with a heterogeneous and extended lifetime of early and late markers at the patches. Additionally, around 50% of the endocytic patches exhibit abnormal spatial dynamics, with immobile patches and patches that bounce backwards to the cell surface, showing a pervasive effect of the absence of AP‐2. These alterations in the endocytic machinery result in abnormal cell wall synthesis and morphogenesis. Our results complement those found in budding yeast and confirm that a direct role of AP‐2 in endocytosis has been conserved throughout evolution. In metazoans the AP‐2 complex has a well‐defined role in clathrin‐mediated endocytosis. By contrast, its direct role in endocytosis in unicellular eukaryotes has been questioned. In S. pombe, apl3Δ mutants exhibit defects in the lifetimes and spatial dynamics of the endocytic patches that lead to alterations in endocytosis, cell wall synthesis, and polarity establishment. These results confirm that a direct role for AP‐2 in endocytosis has been conserved throughout evolution.
ISSN:0950-382X
1365-2958
DOI:10.1111/mmi.13327