Analysis of the Transforming Potential of the Human H-ras Gene by Random Mutagenesis

Some tumor cells contain mutant ras genes that are capable of transforming NIH 3T3 cells. Those genes that have been analyzed arise from the wild-type, non-transforming ras genes by mutations producing single amino acid substitutions at position 12 or 61 of the encoded protein. We have performed ran...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1984-07, Vol.81 (13), p.4006-4010
Main Authors: Fasano, Ottavio, Aldrich, Thomas, Tamanoi, Fuyuhiko, Taparowsky, Elizabeth, Furth, Mark, Wigler, Michael
Format: Article
Language:English
Subjects:
3T3 cells
Amino acid substitution
Amino acids
DNA
Gels
Genetic mutation
Mutagenesis
NIH 3T3 cells
Plasmids
ras genes
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Summary:Some tumor cells contain mutant ras genes that are capable of transforming NIH 3T3 cells. Those genes that have been analyzed arise from the wild-type, non-transforming ras genes by mutations producing single amino acid substitutions at position 12 or 61 of the encoded protein. We have performed random bisulfite-induced mutagenesis on the cloned wild-type human H-ras gene to find if mutations at other positions can activate the transforming potential of that gene. Most mutations are not activating, but mutations that specify single amino acid substitutions at position 12, 13, 59, or 63 of the encoded protein do activate the transforming potential of the H-ras gene. Some, but not all, mutant ras proteins show an altered electrophoretic mobility in NaDodSO4/polyacrylamide gels.
ISSN:0027-8424