A genome-wide RNAi screen identifies multiple RSK-dependent regulators of cell migration.
To define the functional pathways regulating epithelial cell migration, we performed a genome-wide RNAi screen using 55,000 pooled lentiviral shRNAs targeting ∼11,000 genes, selecting for transduced cells with increased motility. A stringent validation protocol generated a set of 31 genes representing diverse pathways whose knockdown dramatically enhances cellular migration. Some of these pathways share features of epithelial-to-mesenchymal transition (EMT), and together they implicate key regulators of transcription, cellular signaling, and metabolism, as well as novel modulators of cellular trafficking, such as DLG5. In delineating downstream pathways mediating these migration phenotypes, we observed universal activation of ERKs and a profound dependence on their RSK effectors. Pharmacological inhibition of RSK dramatically suppresses epithelial cell migration induced by knockdown of all 31 genes, suggesting that convergence of diverse migratory pathways on this kinase may provide a therapeutic opportunity in disorders of cell migration, including cancer metastasis.
Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, 02129, USA.
Gromoslaw A
GA
Smolen
Jianmin
J
Zhang
Matthew J
MJ
Zubrowski
Elena J
EJ
Edelman
Biao
B
Luo
Min
M
Yu
Lydia W
LW
Ng
Cally M
CM
Scherber
Benjamin J
BJ
Schott
Sridhar
S
Ramaswamy
Daniel
D
Irimia
David E
DE
Root
Daniel A
DA
Haber
D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D004847:Epithelial Cells / Q000166:cytology; D055106:Genome-Wide Association Study; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology; D034622:RNA Interference; D015203:Reproducibility of Results; D019893:Ribosomal Protein S6 Kinases / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000378:metabolism
C114461:DLG5 protein, human; D008565:Membrane Proteins; D025521:Tumor Suppressor Proteins; D019893:Ribosomal Protein S6 Kinases