RT Journal Article
SR Electronic
T1 Diverse roles of guanine nucleotide exchange factors in regulating collective cell migration
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 076125
DO 10.1101/076125
A1 Assaf Zaritsky
A1 Yun-Yu Tseng
A1 M. Angeles Rabadan
A1 Shefali Krishna
A1 Michael Overholtzer
A1 Gaudenz Danuser
A1 Alan Hall
YR 2017
UL http://biorxiv.org/content/early/2017/03/14/076125.abstract
AB Efficient collective migration depends on a balance between contractility and cytoskeletal rearrangements, adhesion, and mechanical cell-cell communication, all controlled by GTPases of the RHO family. By comprehensive screening of guanine nucleotide exchange factors (GEFs) in human bronchial epithelial cell monolayers, we identified GEFs that are required for collective migration at large, such as SOS1 and β-PIX, and RHOA GEFs that are implicated in intercellular communication. Downregulation of the latter GEFs differentially enhanced front-to-back propagation of guidance cues through the monolayer, and was mirrored by downregulation of RHOA expression and myosin-II activity. Phenotype-based clustering of knock-down behaviors identified RHOA-ARHGEF18 and ARHGEF3-ARHGEF28-ARHGEF11 clusters, indicating that the latter may signal through other RHO-family GTPases. Indeed, knock-down of RHOC produced an intermediate between the two phenotypes. We conclude that for effective collective migration the RHOA-GEFs-→ARHOA/C→ actomyosin pathways must be optimally tuned to compromise between generation of motility forces and restriction of intercellular communication.GEFguanine nucleotide exchange factorsGDIguanine dissociation inhibitors16HBEhuman bronchial epithelial cells from the 16HBE14o linePCAprincipal component analysisPCprincipal componentsshRNAsmall hairpin RNAKDknockdownFBSfetal bovine serumPCRpolymerase chain reactionqRT-PCRquantitative real time PCRROIregion of interest