DASC, a sensitive classifier for measuring discrete early stages in clathrin-mediated endocytosis [article]

Xinxin Wang, Zhiming Chen, Marcel Mettlen, Jungsik Noh, Sandra L. Schmid, Gaudenz Danuser
2020 bioRxiv   pre-print
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
more » ... nguishing abortive-clathrin 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 unambiguously separates ACs from CCPs without an additional marker. 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 orthogonal to the variation in biochemical measurements of CME. Thus, DASC is an essential tool for uncovering the function of individual EAPs.
doi:10.1101/2020.01.28.924019 fatcat:7yx46arsnrgzrpivjkbw5hohpq