Catabolic control of the enhanced alanine-preferring system for amino acid transport in glucose-starved hamster cells requires protein synthesis

摘要

In cultured hamster cells starved for glucose for 24 hr there is an enhancement of the rate of α-aminoisobutyric acid transport (“shiftup”). When the starved cells are re-fed with glucose, the rate of transport shifts back down to the low, “regulated” rate typical of cells continuously fed with medium containing glucose (“shiftdown”). The high, deregulated rate of transport is maintained, however, when cycloheximide is present for 24 hr during the re-feeding with glucose. Maintenance of the high transport rate is evident only when the cells are incubated in amino acid-free medium just prior to the transport assay or when the assays are conducted with isolated membrane vesicles. A premature, pseudoshiftdown was observed in intact cells within as little as 2 hr after re-feeding when care was not taken to deplete the amino acid pool prior to the transport assay. In addition, a cycloheximide-insensitive increase in transport was observed when cultures were re-fed for 2 hr with amino acid-free medium containing fresh serum. These results emphasize the often overlooked precautions that should be taken to guard against artifacts that could mislead interpretations of amino acid transport data. More important, however, is the finding that Na⁺-dependent amino acid transport in cultured animal cells is regulated in part by a factor (or factors) that becomes inactivated when the cells are maintained under nonglycolytic culture conditions. In order to reactivate the control mechanism, starved cells that have been re-fed with glucose must resynthesize the regulatory factor(s). Thus, in at least cultured hamster cells, Na⁺-dependent amino acid transport regulation is much like the hexose transport regulation in that catabolic control (shiftdown) requires protein synthesis.