TY - JOUR TI - DASC, a sensitive classifier for measuring discrete early stages in clathrin-mediated endocytosis AU - Wang, Xinxin AU - Chen, Zhiming AU - Mettlen, Marcel AU - Noh, Jungsik AU - Schmid, Sandra L AU - Danuser, Gaudenz A2 - Pfeffer, Suzanne R A2 - Geli, María Isabel VL - 9 PY - 2020 DA - 2020/04/30 SP - e53686 C1 - eLife 2020;9:e53686 DO - 10.7554/eLife.53686 UR - https://doi.org/10.7554/eLife.53686 AB - Clathrin-mediated endocytosis (CME) in mammalian cells is driven by resilient machinery that includes >70 endocytic accessory proteins (EAP). Accordingly, perturbation of individual EAPs often results in minor effects on biochemical measurements of CME, thus providing inconclusive/misleading information regarding EAP function. Live-cell imaging can detect earlier roles of EAPs preceding cargo internalization; however, this approach has been limited because unambiguously distinguishing abortive coats (ACs) from bona fide clathrin-coated pits (CCPs) is required but unaccomplished. Here, we develop a thermodynamics-inspired method, “disassembly asymmetry score classification (DASC)”, that resolves ACs from CCPs based on single channel fluorescent movies. After extensive verification, we use DASC-resolved ACs and CCPs to quantify CME progression in 11 EAP knockdown conditions. We show that DASC is a sensitive detector of phenotypic variation in CCP dynamics that is uncorrelated to the variation in biochemical measurements of CME. Thus, DASC is an essential tool for uncovering EAP function. KW - endocytosis KW - unbiased classification KW - endocytic accessory proteins JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -