title: Expression of Gab1 lacking the pleckstrin homology domain is associated with neoplastic progression.
abstract: An in vitro transformation system of carcinogen-treated Syrian hamster embryo (SHE) cell cultures represents multistep genetic and nongenetic changes that develop during the neoplastic progression of normal cells to tumor cells in vivo. During this neoplastic progression, SHE cells demonstrate an altered response to epidermal growth factor (EGF). In the present report, we examined the role of the adapter protein Gab1 (Grb2-associated binder-1) in the neoplastic progression of SHE cells. We used two asbestos-transformed SHE cell clones in different neoplastic stages: a 10W+8 clone, which is immortal and retains the ability to suppress the tumorigenicity of tumor cells in cell-cell hybrid experiments, and a 10W-1 clone, which has lost this tumor suppressor ability. 10W+8 cells expressed full-length 100-kDa Gab1 and associated 5.2-kb mRNA. Upon repeated cell passaging, 10W-1 cells showed increasing expression of a novel 87-kDa form of Gab1 as well as 4.6-kb mRNA with diminishing expression of the original 100-kDa Gab1. cDNA encoding the 87-kDa Gab1 predicts a form of Gab1 lacking the amino-terminal 103 amino acids (Gab1(Delta1-103)), which corresponds to loss of most of the pleckstrin homology (PH) domain. Gab1(Delta1-103) retains the ability to be phosphorylated in an EGF-dependent manner and to associate with the EGF receptor and SHP-2 upon EGF stimulation. The endogenous expression of Gab1(Delta1-103) in 10W-1 cells appeared closely related to EGF-dependent colony formation in soft agar. Moreover, transfection and expression of Gab1(Delta1-103), but not Gab1, in 10W+8 cells enhanced their EGF-dependent colony formation in soft agar. These results demonstrate that Gab1 is a target of carcinogen-induced transformation of SHE cells and that the expression of a Gab1 variant lacking most of the PH domain plays a specific role in the neoplastic progression of SHE cells.
authors.affiliation: Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709, USA.
authors.forename: H
authors.initials: H
authors.lastname: Kameda
authors.forename: J I
authors.initials: JI
authors.lastname: Risinger
authors.forename: B B
authors.initials: BB
authors.lastname: Han
authors.forename: S J
authors.initials: SJ
authors.lastname: Baek
authors.forename: J C
authors.initials: JC
authors.lastname: Barrett
authors.forename: T
authors.initials: T
authors.lastname: Abe
authors.forename: T
authors.initials: T
authors.lastname: Takeuchi
authors.forename: W C
authors.initials: WC
authors.lastname: Glasgow
authors.forename: T E
authors.initials: TE
authors.lastname: Eling
mesh_terms: D000362:Agar / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001194:Asbestos; D001483:Base Sequence; D001798:Blood Proteins / Q000737:chemistry*; D015152:Blotting, Northern; D015153:Blotting, Western; D002273:Carcinogens; D002455:Cell Division; D002460:Cell Line; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000378:metabolism; D018450:Disease Progression; D004815:Epidermal Growth Factor / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D020935:MAP Kinase Signaling System; D008647:Mesocricetus; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000139:chemically induced / Q000378:metabolism*; D010641:Phenotype; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000737:chemistry*; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D014170:Transformation, Genetic
chemical_list: D001798:Blood Proteins; D002273:Carcinogens; D018076:DNA, Complementary; D010750:Phosphoproteins; D012333:RNA, Messenger; C039077:platelet protein P47; D001194:Asbestos; D004815:Epidermal Growth Factor; D000362:Agar