%0 Journal Article %A Morgane Rosendale %A Thi Nhu Ngoc Van %A Dolors Grillo-Bosch %A Silvia Sposini %A Léa Claverie %A Isabel Gauthereau %A Stéphane Claverol %A Daniel Choquet %A Matthieu Sainlos %A David Perrais %T Functional recruitment of dynamin requires multimeric interactions for efficient endocytosis %D 2019 %R 10.1101/309245 %J bioRxiv %P 309245 %X During clathrin mediated endocytosis (CME), membrane scission is achieved by the concerted action of dynamin and its interacting partners. Essential interactions occur between the proline/arginine-rich domain of dynamin (dynPRD) and the Src-homology domain 3 (SH3) of various proteins including amphiphysins. Here we show that multiple SH3 domains must bind simultaneously to dynPRD through three adjacent motifs for dynamin’s efficient recruitment and function. First, we show in dynamin triple knock-out cells that mutant dynamins modified in a single motif, including the central amphiphysin SH3 (amphSH3) binding motif, are partially capable of rescuing CME. However, mutating two motifs largely prevents that ability. To support this observation, we designed divalent dynPRD-derived peptides. These ligands bind multimers of amphSH3 with >100-fold higher affinity than monovalent ones in vitro. Accordingly, dialyzing living cells with these divalent peptides through a patch-clamp pipette blocks CME 2 to 3 times more effectively than with monovalent ones. Finally, the frequency of endocytic events decreases with competing peptides or hypomorphic rescue mutants but the kinetics of dynamin recruitment is unaffected. This suggests that PRD-SH3 interactions act upstream of dynamin accumulation at the neck of nascent vesicles. We conclude from these data that dynamin drives vesicle scission via multivalent interactions in vivo. %U https://www.biorxiv.org/content/biorxiv/early/2019/07/30/309245.full.pdf