Role of idle ribosomes in the response of Chinese hamster ovary cells to depletion of histidyl-tRNA.
In Chinese hamster ovary cells, histidine starvation and inactivation of histidyl-tRNA synthetase by mutations or histidinol result in stimulation of protein breakdown. We have previously shown that the regulatory mechanism recognizes the level of aminoacylation of tRNA(His). We now report that it is also sensitive to the functional state of the ribosomes. Cycloheximide, an inhibitor of peptidyl-tRNA translocation, decreases the sensitivity of the regulation. In the presence of 1.5 micrograms cycloheximide/ml, protein synthesis is inhibited to 6% of control; a full response can still be elicited by appropriate concentrations of histidinol, but it requires a more extensive depletion of histidyl-tRNA than in the absence of cycloheximide. The response is attained only when the depletion is sufficient to inhibit protein synthesis further and to increase the number of ribosomes idling in the histidine codon with an empty aminoacyl site, measured by their reactivity in vivo to low concentrations of puromycin. The results indicate that a simple depletion of his-tRNA is not sufficient to elicit the response and suggest that idle ribosomes are required for regulation.
Scornik
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Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03756.
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003513:Cycloheximide / Q000494:pharmacology; D006641:Histidinol / Q000494:pharmacology; D009154:Mutation; D011691:Puromycin / Q000494:pharmacology; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D012346:RNA, Transfer, Amino Acyl; D011691:Puromycin; D006641:Histidinol; D003513:Cycloheximide