Spatio-temporal regulation of endocytic protein assembly by SH3 domains in yeast

• Published in: Molecular biology of the cell 2023-03, Vol.34 (3), p.ar19
• Main Authors: Hummel, Daniel R, Kaksonen, Marko
• Format: Article
• Language: English
• Subjects: Actins - metabolism; Endocytosis - physiology; Microfilament Proteins - metabolism; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - metabolism; src Homology Domains; Wiskott-Aldrich Syndrome Protein - metabolism
• ISSN: 1059-1524; 1939-4586
• DOI: 10.1091/mbc.E22-09-0406

Clathrin-mediated endocytosis is a conserved eukaryotic membrane trafficking pathway that is driven by a sequentially assembled molecular machinery that contains over 60 different proteins. SH3 domains are the most abundant protein-protein interaction domain in this process, but the function of most SH3 domains in protein dynamics remains elusive. Using mutagenesis and live-cell fluorescence microscopy in the budding yeast , we dissected SH3-mediated regulation of the endocytic pathway. Our data suggest that multiple SH3 domains regulate the actin nucleation-promoting Las17-Vrp1 complex, and that the network of SH3 interactions coordinates both Las17-Vrp1 assembly and dissociation. Furthermore, most endocytic SH3 domain proteins use the SH3 domain for their own recruitment, while a minority use the SH3 domain to recruit other proteins and not themselves. Our results provide a dynamic map of SH3 functions in yeast endocytosis and a framework for SH3 interaction network studies across biology.