High-level expression of human c-Src can cause a spherical morphology without loss of anchorage-dependent growth of NIH 3T3 cells.
To investigate whether overexpression of human c-Src leads to cell rounding in anchorage-dependent NIH 3T3 fibroblasts, we have established c-Src-inducible cell lines using a lac repressor-operator system. RN1 cells, which expressed c-Src at a high level after induction, exhibited a spherical morphology and ceased to grow in monolayer culture. RN1 cells, however, exhibited neither focus-forming ability nor anchorage-independent growth potential with or without induction. Induced RN1 c-Src was phosphorylated at Ser75, a previously reported spherical cell-associated site, and at Tyr419. These data demonstrated for the first time that highly elevated human c-Src tyrosine kinase activity can cause NIH 3T3 cells to have a spherical morphology without loss of anchorage-dependent growth. The inducible cell line should be useful to study the mechanism for cell rounding by c-Src.
Kato
G
G
Department of Biochemistry, Yamanashi Medical University, Nakakoma, Japan. gkato@res.yamanashi-med.ac.jp
Maeda
S
S
D016475:3T3 Cells; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002448:Cell Adhesion / Q000235:genetics; D002455:Cell Division; D048429:Cell Size; D004591:Electrophoresis, Polyacrylamide Gel; D029968:Escherichia coli Proteins; D005786:Gene Expression Regulation / Q000235:genetics*; D016391:Genes, src; D006801:Humans; D015151:Immunoblotting; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D056968:Lac Repressors; D051379:Mice; D010449:Peptide Mapping; D010748:Phosphopeptides / Q000032:analysis / Q000737:chemistry; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D011233:Precipitin Tests; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics; D014162:Transfection / Q000235:genetics
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D056968:Lac Repressors; D010748:Phosphopeptides; D012097:Repressor Proteins; D010768:Phosphoserine; D007544:Isopropyl Thiogalactoside; D016392:Proto-Oncogene Proteins pp60(c-src)