Gene trapping identifies inhibitors of oncogenic transformation. The tissue inhibitor of metalloproteinases-3 (TIMP3) and collagen type I alpha2 (COL1A2) are epidermal growth factor-regulated growth repressors.
A gene trap strategy has been used to identify genes that are repressed in cells transformed by an activated epidermal growth factor (EGF)/EGF receptor signal transduction pathway. EGF receptor-expressing NIH3T3 cells (HER1 cells) were infected with a retrovirus containing coding sequences for the human CD2 antigen and for secreted alkaline phosphatase in the U3 region. By selecting for and against CD2 expression, we obtained clones in which the gene trap had integrated into genes selectively repressed by EGF. Two of these clones encoded for the secreted extracellular matrix proteins TIMP3 and COL1A2. We show here that both genes are downstream targets of RAS and are specifically repressed by EGF-induced transformation. Moreover, this strategy tags tumor suppressor genes in their normal chromosomal location, thereby improving target-specific screens for antineoplastic drugs.
Andreú
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T
Laboratory for Molecular Hematology, Department of Hematology, University of Frankfurt Medical School, Weismüllerstrasse 45, Frankfurt am Main, Germany.
Beckers
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Thoenes
E
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Hilgard
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P
von Melchner
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H
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D003094:Collagen / Q000235:genetics*; D004247:DNA; D004815:Epidermal Growth Factor / Q000235:genetics* / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics*
D012333:RNA, Messenger; D019717:Tissue Inhibitor of Metalloproteinase-3; D004815:Epidermal Growth Factor; D003094:Collagen; D004247:DNA; D066246:ErbB Receptors