title: Dynamin triple knockout cells reveal off target effects of commonly used dynamin inhibitors.
abstract: Dynamin, which is encoded by three genes in mammals, is a GTPase implicated in endocytic membrane fission. Dynamin 1 and 3 are predominantly expressed in brain, whereas dynamin 2 is ubiquitously expressed. With the goal of assessing the impact of the lack of dynamin on cell physiology, we previously generated and characterized dynamin 1 and 2 double knockout (DKO) fibroblasts. These DKO cells were unexpectedly viable in spite of a severe impairment of clathrin-mediated endocytosis. As low-level expression of the dynamin 3 gene in these cells could not be excluded, we have now engineered dynamin 1, 2 and 3 triple KO (TKO) fibroblasts. These cells did not reveal any additional defects beyond what was previously observed in DKO fibroblasts. Surprisingly, although fluid-phase endocytosis and peripheral membrane ruffling were not impaired by the lack of all three dynamins, two structurally similar, widely used dynamin inhibitors, dynasore and Dyngo-4a, robustly inhibited these two processes both in wild-type and TKO cells. Dynamin TKO cells will be useful tools for the further exploration of dynamin-dependent processes and the development of more specific dynamin inhibitors.
authors.affiliation: Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA.
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: Park
authors.forename: Hongying
authors.initials: H
authors.lastname: Shen
authors.forename: Lijuan
authors.initials: L
authors.lastname: Liu
authors.forename: Xinran
authors.initials: X
authors.lastname: Liu
authors.forename: Shawn M
authors.initials: SM
authors.lastname: Ferguson
authors.forename: Pietro
authors.initials: P
authors.lastname: De Camilli
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D034284:Dynamin I / Q000235:genetics / Q000378:metabolism*; D034285:Dynamin II / Q000235:genetics / Q000378:metabolism*; D034286:Dynamin III / Q000235:genetics / Q000378:metabolism*; D004705:Endocytosis / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D055786:Gene Knockout Techniques; D006835:Hydrazones / Q000494:pharmacology; D051379:Mice; D009284:Naphthols / Q000494:pharmacology; D013569:Synapses / Q000378:metabolism*
chemical_list: D006835:Hydrazones; C511794:N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide; D009284:Naphthols; C568360:dyngo-4a; D034284:Dynamin I; D034285:Dynamin II; D034286:Dynamin III
keywords: Actin; Dynamin; Dynasore; Dyngo; Synapse; Synaptic vesicle
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