Expression of Rous sarcoma virus transforming protein pp60v-src in Saccharomyces cerevisiae cells.

J. S. Brugge, G. Jarosik, J. Andersen, A. Queral-Lustig, M. Fedor-Chaiken, James Broach

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Abstract

The Rous sarcoma virus (RSV) pp60v-src protein was expressed in Saccharomyces cerevisiae cells either from a plasmid vector carrying the v-src gene or in yeast cells containing a single-copy v-src gene chromosomally integrated. In both yeast strains, v-src gene transcription is regulated by the galactose-inducible GAL10 promoter. Growth in galactose-containing medium resulted in constitutive expression of pp60v-src in the integrated strain and transient expression of higher levels of pp60v-src in the plasmid-bearing strain. The concentration of pp60v-src in the plasmid-bearing strain at its peak of expression was approximately threefold lower than that found in RSV-transformed mammalian cells. pp60v-src synthesized in yeast cells was phosphorylated in vivo on sites within the amino and carboxyl halves of the molecule. In immune complex kinase assays, the yeast pp60v-src was autophosphorylated on tyrosine and was able to phosphorylate exogenous substrates such as casein and enolase. The specific activity of pp60v-src synthesized in yeast cells was approximately 5- to 10-fold higher than that made in mammalian cells. Induction of pp60v-src caused the death of the plasmid-bearing yeast strain and transient inhibition of growth of the single-copy strain. Concomitantly, this induction resulted in high levels of tyrosine phosphorylation of yeast cell proteins. This indicates that pp60v-src functions as a tyrosine-specific phosphotransferase in yeast cells and suggests that hyperphosphorylation of yeast proteins is inimical to cell growth.

Original languageEnglish (US)
Pages (from-to)2180-2187
Number of pages8
JournalMolecular and Cellular Biology
Volume7
Issue number6
DOIs
StatePublished - Jan 1 1987

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Rous sarcoma virus
Saccharomyces cerevisiae
Yeasts
src Genes
Proteins
Plasmids
Tyrosine
Fungal Proteins
Galactose
Phosphotransferases
Growth
Phosphopyruvate Hydratase
Caseins
Antigen-Antibody Complex
Phosphorylation

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Brugge, J. S. ; Jarosik, G. ; Andersen, J. ; Queral-Lustig, A. ; Fedor-Chaiken, M. ; Broach, James. / Expression of Rous sarcoma virus transforming protein pp60v-src in Saccharomyces cerevisiae cells. In: Molecular and Cellular Biology. 1987 ; Vol. 7, No. 6. pp. 2180-2187.
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Expression of Rous sarcoma virus transforming protein pp60v-src in Saccharomyces cerevisiae cells. / Brugge, J. S.; Jarosik, G.; Andersen, J.; Queral-Lustig, A.; Fedor-Chaiken, M.; Broach, James.

In: Molecular and Cellular Biology, Vol. 7, No. 6, 01.01.1987, p. 2180-2187.

Research output: Contribution to journalArticle

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T1 - Expression of Rous sarcoma virus transforming protein pp60v-src in Saccharomyces cerevisiae cells.

AU - Brugge, J. S.

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