RNAi-mediated Hip1R Silencing Results in Stable Association between the Endocytic Machinery and the Actin Assembly Machinery

Åsa E.Y. Engqvist-Goldstein, Claire X. Zhang, Sebastien Carreno, Consuelo Barroso, John E. Heuser, David G. Drubin*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

136 Citations (Scopus)


Actin filaments transiently associate with the endocytic machinery during clathrin-coated vesicle formation. Although several proteins that might mediate or regulate this association have been identified, in vivo demonstration of such an activity has not been achieved. Huntingtin interacting protein 1R (Hip1R) is a candidate cytoskeletal-endocytic linker or regulator because it binds to clathrin and actin. Here, Hip1R levels were lowered by RNA interference (RNAi). Surprisingly, rather than disrupting the transient association between endocytic and cytoskeletal proteins, clathrin-coated structures (CCSs) and their endocytic cargo became stably associated with dynamin, actin, the Arp2/3 complex, and its activator, cortactin. RNAi double-depletion experiments demonstrated that accumulation of the cortical actin-endocytic complexes depended on cortactin. Fluorescence recovery after photobleaching showed that dynamic actin filament assembly can occur at CCSs. Our results provide evidence that Hip1R helps to make the interaction between actin and the endocytic machinery functional and transient.

Original languageEnglish
Pages (from-to)1666-1679
Number of pages14
JournalMolecular Biology of the Cell
Issue number4
Publication statusPublished - Apr 2004
Externally publishedYes

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