Transformation of Chicken Embryo Fibroblast Cells by Avian Retroviruses Containing the Human Fyn Gene and Its Mutated Genes

Kentaro Senba, Sadaaki Kawai, Yumiko Matsuzawa, Yuji Yamanashi, Makoto Nishizawa, Kumao Toyoshima

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The transforming activity of the human fyn protein, p59fyn, which is a kinase of the src family, was investigated by testing the effect of recombinant avian retrovirus (Fyn virus) expressing p59fyn on chickens or cultured chicken embryo fibroblast (CEF) cells. The Fyn virus did not induce transformed foci. After several passages of the virus stock on CEF cells, however, a few foci were detected in the presence of dimethyl sulfoxide. Chickens inoculated with Fyn virus at the stage of 12-day-old embryos developed fibrosarcomas 3 to 6 weeks after hatching. The viruses obtained from these foci and from one of the tumor tissues showed high transforming activity in the presence of dimethyl sulfoxide, suggesting that these viruses carry spontaneous mutations of the fyn gene. Four fyn genes from CEF DNAs infected with transforming viruses were molecularly cloned, and their products were confirmed to possess transforming activity. DNA sequence analysis of the fyn genes showed that two of the four mutants have Thr instead of Ile at position 338 in the kinase domain. The other two mutants carry deletions of 78 and 108 base pairs, respectively, which result in complete loss of region C of SH2. The overall level of proteins containing phosphotyrosine was significantly higher in transformed cells than in normal CEF cells. Our data indicate that when expressed at high levels in a retrovirus, normal p59fyn cannot cause cellular transformation, but that mutant p59fyn with either a single amino acid substitution in the kinase domain or a deletion including region C produces a transforming protein, perhaps due to enhanced tyrosine kinase activity. This is the first observation that deletion of region C can unmask the potential transforming activity of a src family kinase.

Original languageEnglish
Pages (from-to)3095-3104
Number of pages10
JournalMolecular and Cellular Biology
Volume10
Issue number6
Publication statusPublished - 1990 Jun
Externally publishedYes

Fingerprint

Retroviridae
Chickens
Embryonic Structures
Fibroblasts
Viruses
Genes
src-Family Kinases
Dimethyl Sulfoxide
Phosphotransferases
Phosphotyrosine
Proteins
Fibrosarcoma
Amino Acid Substitution
DNA Sequence Analysis
Human Activities
Base Pairing
Protein-Tyrosine Kinases
Observation
Mutation
DNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Transformation of Chicken Embryo Fibroblast Cells by Avian Retroviruses Containing the Human Fyn Gene and Its Mutated Genes. / Senba, Kentaro; Kawai, Sadaaki; Matsuzawa, Yumiko; Yamanashi, Yuji; Nishizawa, Makoto; Toyoshima, Kumao.

In: Molecular and Cellular Biology, Vol. 10, No. 6, 06.1990, p. 3095-3104.

Research output: Contribution to journalArticle

Senba, K, Kawai, S, Matsuzawa, Y, Yamanashi, Y, Nishizawa, M & Toyoshima, K 1990, 'Transformation of Chicken Embryo Fibroblast Cells by Avian Retroviruses Containing the Human Fyn Gene and Its Mutated Genes', Molecular and Cellular Biology, vol. 10, no. 6, pp. 3095-3104.
Senba, Kentaro ; Kawai, Sadaaki ; Matsuzawa, Yumiko ; Yamanashi, Yuji ; Nishizawa, Makoto ; Toyoshima, Kumao. / Transformation of Chicken Embryo Fibroblast Cells by Avian Retroviruses Containing the Human Fyn Gene and Its Mutated Genes. In: Molecular and Cellular Biology. 1990 ; Vol. 10, No. 6. pp. 3095-3104.
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