title: RNAi-based functional selection identifies novel cell migration determinants dependent on PI3K and AKT pathways.
abstract: Lentiviral short hairpin RNA (shRNA)-mediated genetic screening is a powerful tool for identifying loss-of-function phenotype in mammalian cells. Here, we report the identification of 91 cell migration-regulating genes using unbiased genome-wide functional genetic selection. Individual knockdown or cDNA overexpression of a set of 10 candidates reveals that most of these cell migration determinants are strongly dependent on the PI3K/PTEN/AKT pathway and on their downstream signals, such as FOXO1 and p70S6K1. ALK, one of the cell migration promoting genes, uniquely uses p55γ regulatory subunit of PI3K, rather than more common p85 subunit, to trigger the activation of the PI3K-AKT pathway. Our method enables the rapid and cost-effective genome-wide selection of cell migration regulators. Our results emphasize the importance of the PI3K/PTEN/AKT pathway as a point of convergence for multiple regulators of cell migration.
authors.affiliation: 1] Department of Pharmacology, Brain Science &Engineering Institute, BK21 Plus KNU Biomedical Convergence Program, Kyungpook National University School of Medicine, Daegu, Republic of Korea [2] College of Medicine, Dongguk University, Gyeongju, Republic of Korea.
authors.forename: Minchul
authors.initials: M
authors.lastname: Seo
authors.affiliation: 1] Department of Pharmacology, Brain Science &Engineering Institute, BK21 Plus KNU Biomedical Convergence Program, Kyungpook National University School of Medicine, Daegu, Republic of Korea [2] Korea Brain Research Institute (KBRI), Daegu, Republic of Korea.
authors.forename: Shinrye
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Pharmacology, Brain Science &Engineering Institute, BK21 Plus KNU Biomedical Convergence Program, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
authors.forename: Jong-Heon
authors.initials: JH
authors.lastname: Kim
authors.affiliation: KNU Creative BioResearch Group, School of Life Sciences and Biotechnology, Kyungpook National University, Daegu, Republic of Korea.
authors.forename: Won-Ha
authors.initials: WH
authors.lastname: Lee
authors.affiliation: Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health and Sciences, Research Triangle Park, North Carolina 27709, USA.
authors.forename: Guang
authors.initials: G
authors.lastname: Hu
authors.affiliation: Department of Genetics, Howard Hughes Medical Institute, Division of Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Elledge
authors.affiliation: Department of Pharmacology, Brain Science &Engineering Institute, BK21 Plus KNU Biomedical Convergence Program, Kyungpook National University School of Medicine, Daegu, Republic of Korea.
authors.forename: Kyoungho
authors.initials: K
authors.lastname: Suk
mesh_terms: D000077548:Anaplastic Lymphoma Kinase; D000818:Animals; D002465:Cell Movement; D004789:Enzyme Activation; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D016678:Genome; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D041681:NIH 3T3 Cells; D051059:PTEN Phosphohydrolase / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D021122:Protein Subunits / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism; D015203:Reproducibility of Results; D015398:Signal Transduction; D014162:Transfection
chemical_list: D021122:Protein Subunits; D034741:RNA, Small Interfering; D019869:Phosphatidylinositol 3-Kinases; C000626173:ALK protein, human; C000626174:Alk protein, mouse; D000077548:Anaplastic Lymphoma Kinase; D020794:Receptor Protein-Tyrosine Kinases; D051057:Proto-Oncogene Proteins c-akt; D051059:PTEN Phosphohydrolase