Identification of novel cell migration-promoting genes by a functional genetic screen.
Here we describe the identification of novel cell migration-promoting genes based on an unbiased functional genetic screen in cultured cells. After the introduction of the retroviral mouse brain cDNA library into NIH3T3 mouse fibroblast cells, migration-promoted cells were selected by a 3-dimensional migration assay using cell culture inserts. After 5 rounds of enrichment, cDNAs were retrieved from the cells with a selected phenotype. Cell migration-promoting activity was confirmed by independent migration assays for the retrieved cDNAs, among which further investigation was focused on coiled-coil-helix-coiled-coil-helix domain-containing protein 2 (chchd2). Whereas overexpression of chchd2 promoted cell migration, knockdown of endogenous chchd2 expression reduced cell migration. Chchd2-induced cell migration was associated with augmented formation of actin stress fibers and focal adhesion, which was mediated through Akt, RhoA/ROCK, and Jnk pathways. CHCHD2 protein directly interacted with hyaluronic acid-binding protein 1 (HABP1) that possessed migration-suppressing activity. Intracellular localization and further functional studies suggested that CHCHD2 and HABP1 may mutually regulate each other to balance cell migration. Thus, chchd2 is a novel cell migration determinant identified by an in vitro functional genetic selection strategy. The selection method can also be useful for the isolation of genes that give other phenotypes of interest.
Seo
Minchul
M
Department of Pharmacology, School of Medicine, Brain Science and Engineering Institute, Kyungpook National University, Daegu, Korea.
Lee
Won-Ha
WH
Suk
Kyoungho
K
D000199:Actins / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000502:physiology; D002465:Cell Movement / Q000235:genetics*; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins; D022001:Focal Adhesions / Q000502:physiology; D015723:Gene Library; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics* / Q000502:physiology; D048031:JNK Mitogen-Activated Protein Kinases / Q000502:physiology; D051379:Mice; D024101:Mitochondrial Proteins; D041681:NIH 3T3 Cells; D051057:Proto-Oncogene Proteins c-akt / Q000502:physiology; D015398:Signal Transduction; D014157:Transcription Factors; D014162:Transfection; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
D000199:Actins; C490193:C1qbp protein, mouse; D002352:Carrier Proteins; C547271:Chchd2 protein, mouse; D004268:DNA-Binding Proteins; D018960:Hyaluronan Receptors; D024101:Mitochondrial Proteins; D014157:Transcription Factors; D051057:Proto-Oncogene Proteins c-akt; D048031:JNK Mitogen-Activated Protein Kinases; D020742:rhoA GTP-Binding Protein