Advances in analysis of low signal-to-noise images link dynamin and AP2 to the functions of an endocytic checkpoint

François Aguet, CostinN Antonescu, Marcel Mettlen, SandraL Schmid, Gaudenz Danuser

Research output: Contribution to journalArticlepeer-review

183 Scopus citations

Abstract

Numerous endocytic accessory proteins (EAPs) mediate assembly and maturation of clathrin-coated pits (CCPs) into cargo-containing vesicles. Analysis of EAP function through bulk measurement of cargo uptake has been hampered due to potential redundancy among EAPs and, as we show here, the plasticity and resilience of clathrin-mediated endocytosis (CME). Instead, EAP function is best studied by uncovering the correlation between variations in EAP association to individual CCPs and the resulting variations in maturation. However, most EAPs bind to CCPs in low numbers, making the measurement ofEAP association via fused fluorescent reporters highly susceptible to detection errors. Here, we present a framework for unbiased measurement of EAP recruitment to CCPs and their direct effects on CCP dynamics. We identify dynamin and the EAP-binding α-adaptin appendage domain of the AP2 adaptor as switches in a regulated, multistep maturation process and provide direct evidence for a molecular checkpoint in CME.

Original languageEnglish (US)
Pages (from-to)279-291
Number of pages13
JournalDevelopmental cell
Volume26
Issue number3
DOIs
StatePublished - Aug 12 2013

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Developmental Biology
  • Cell Biology

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