Regulation of hypoxanthine transport by cyclic nucleotides in cultured Chinese hamster lung fibroblasts

Abstract

The regulation of hypoxanthine transport activity by Chinese hamster lung fibroblasts grown in culture was examined in wild-type clones and 8-azaguanine-resistant mutant clones which lack hypoxanthine-guanine phosphoribosyltransferase. Hypoxanthine transport activity increases with increased rates of cellular growth expressed as viable cell number, total cell protein, and DNA synthesis. The transport activity for hypoxanthine declines when the fibroblasts approach confluence or after exposure to cycloheximide or actinomycin D. In vivo incubation of either fibroblast subline with 100 μ m dibutyryl cyclic AMP decreases transport activity over 50%, whereas exposure to 10 μ m dibutyryl cyclic GMP increases hypoxanthine uptake by 40%. A synergistic effect is observed when fibroblasts are incubated with a phosphodiesterase inhibitor (3-isobutyl-1-methylxanthine or theophylline) plus glucagon, an adenylate cyclase stimulator. Such additions result in a 70% decrease in the cellular transport capacity. Stimulation of hypoxanthine transport by 40% is observed following incubation with insulin. Addition of all agents produces maximum changes in the rate of hypoxanthine transport only after a 6-h in vivo incubation with the fibroblasts. These findings suggest that hypoxanthine transport is regulated by the intracellular concentration of cyclic nucleotides. This control may occur at the level of gene expression for a hypoxanthine transport protein.