The effect of amino acid analogues and heat shock on gene expression in chicken embryo fibroblasts

Abstract

The addition of certain amino acid analogues (canavanine, hydroxynorvaline, o-methylthreonine) or a mild heat shock at 45°C caused chicken embryo fibroblasts to increase rapidly the synthesis of three proteins (molecular weights 22,000, 76,000 and 95,000 daltons) to levels which dominate the cells biosynthetic capacity and exceed the level of synthesis of the major cell structural proteins. Actinomycin D blocked the increased synthesis of p22, p76 and p95 in both analogue and heat shock-treated cells, while cycloheximide addition during the “induction” period blocked formation of these proteins only in analogue-treated cells. The elevated levels of synthesis for this set of proteins began to decrease shortly after restoration of the normal amino acid or normal temperature, and the normal pattern of cell protein synthesis was found 8 hr later. Induction of a similar set of proteins was detected in mouse L cells and baby hamster kidney cells after treatment with amino acid analogues or heat shock. Several laboratories have reported synthesis of proteins with similar molecular weights in cells subjected to conditions that alter glucose metabolism, and we speculate that these proteins may be associated with a hexose transport system.