1. The trimeric coiled‐coil HSBP1 protein promotes WASH complex assembly at centrosomes
The stable MDA‐MB‐231 cell line expressing FLAG‐HaloTag‐WASH (FHT‐WASH) was analyzed by Western blot. Overexpression of tagged WASH induces upregulation of CCDC53, whereas FAM21 and Strumpellin levels are not affected. Tagged WASH replaces the endogenous WASH in the WASH complex. Stable MDA‐MB‐231 cells expressing FHT‐WASH were treated for 24 h with HaloPROTAC3, a small molecule that degrades HaloTag tagged proteins (Buckley et al, ). FHT‐WASH degradation induced CCDC53 downregulation and reappearance of endogenous WASH. FAM21 and Strumpellin levels were not affected. HaloPROTAC3 wash‐out for the indicated time allows to monitor subunit buildup around FHT‐WASH by FLAG immunoprecipitations. Upon HSBP1 depletion, the association of WASH with CCDC53 is delayed. Source data are available online for this figure.
IF THIS IMAGE HAS BEEN PROVIDED BY OR IS OWNED BY A THIRD PARTY, AS INDICATED IN THE CAPTION LINE, THEN FURTHER PERMISSION MAY BE NEEDED BEFORE ANY FURTHER USE. PLEASE CONTACT WILEY'S PERMISSIONS DEPARTMENT ON OR USE THE RIGHTSLINK SERVICE BY CLICKING ON THE 'REQUEST PERMISSIONS' LINK ACCOMPANYING THIS ARTICLE. WILEY OR AUTHOR OWNED IMAGES MAY BE USED FOR NON-COMMERCIAL PURPOSES, SUBJECT TO PROPER CITATION OF THE ARTICLE, AUTHOR, AND PUBLISHER.
EMBO J, Volume: 37, Issue: 13, First published: 29 May 2018, DOI: ( /embj )