Multiplexed quantitative screens of single cell shape and YAP/TAZ localisation identify DOCK5 as a coincident detector of polarity and adhesion during migration

Abstract

YAP and TAZ are transcriptional co-activators that are often constitutively active in triple negative breast cancer (TNBC) cells. However the mechanisms responsible for YAP/TAZ dysregulation in TNBC remain unclear. We hypothesised that RhoGEF activity in TNBC cells underpins YAP/TAZ activation in TNBC cells. Through systematic RNAi screening we found that the Focal Adhesion (FA) associated RhoGEF DOCK5 promotes YAP/TAZ nuclear translocation in proliferating TNBC cells. DOCK5 is required for 2D migration and 3D invasion, acting as a coincident detector that maintains cell polarity by stabilising GSK3β activation downstream of CDC42 at polarised leading edges only where FAs are being assembled. DOCK5 and CDC42 are required for cell survival and drug resistance in TNBC cells. Based on these studies we propose that cancer cell phenotypes such as drug resistance can be driven by cells adopting polarised, migratory shapes.

A Common Genetic Factor Underlies Genetic Risk for Gynaecological and Reproductive Disorders and Is Correlated with Risk to Depression

“…Here, the importance of rs11031006 and its associated genes, FSHB and ARL14EP, was highlighted by its most highly significant association with the latent reproductive factor. FSHB, as well as GREB1, ESR1, SYNE1, WT1, WNT4, and TERT, whose effect is largely mediated through this latent reproductive factor, have a strong association with reproductive health [2,[65][66][67][68][69][70], while RUNX1, RNLS, DOCK5, and EEFSEC promote genome stability [71][72][73][74][75].…”

Nuclear rupture induced by capillary constriction forces promotes differential effects on metastatic and normal breast cells

“…The invasion assay was performed with ECM gel from Engelbreth-Holm-Swarm murine sarcoma (Sigma-Aldrich, E6909). Ultra low attachment plates ULA (Greiner Bio-one, 650979) were used for the production of spheroids following the protocol from Pascual-Vargas et al 70 . DMEM 5× was prepared using 12.5 g DMEM powder, 25 ml of 1 M HEPES pH 7.5, 5 g NaHCO 3 , H 2 O up to 250 ml and was filtered under sterile conditions before aliquoting and freezing it at − 20 °C.…”