Control of protein synthesis by extracellular Na+ in cultured fibroblasts.
In chick embryo fibroblasts (CEFs), a partial substitution of extracellular Na+ with other cations or carbohydrates decreased the intracellular Na+ content without altering the K+ level. Concomitantly, a significant decrease in the serum-dependent rate of protein synthesis occurred. This phenomenon appeared to be quickly reversible upon reconstitution of the correct extracellular Na+ concentration in the culture medium. The presence of a transcriptional inhibitor such as actinomycin D during the treatment did not inhibit the reversibility of the phenomenon. The presence in the culture medium of K+ in such excess as to dissipate the membrane potential did not alter the observed relationship between the protein synthesis rate and the internal Na+ content. Analysis of the amino acid pool indicated that the observed inhibition of the rate of protein synthesis in CEFs incubated in low Na+ medium was not caused by an unbalanced availability of intracellular amino acids. In addition, intracellular pH, as estimated by the measurement of the equilibrium distribution of benzoic acid, did not show any significant alteration in cells incubated in the presence of bicarbonate buffer and in low extracellular Na+. Moreover, the relationship between the rate of protein synthesis and the internal Na+ content was still observed in CEFs cultured in bicarbonate-containing media, but at lower or higher than physiological pH. Analysis by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of the proteins synthesized by CEFs cultured at a reduced extracellular Na+ concentration showed that specific alterations of gene expression occurred.
Petronini
P G
PG
Istituto di Patologia Generale, Università di Parma, Italy.
Tramacere
M
M
Mazzini
A
A
Kay
J E
JE
Borghetti
A F
AF
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000494:pharmacology; D002412:Cations / Q000494:pharmacology; D002478:Cells, Cultured; D002642:Chick Embryo; D015180:Electrophoresis, Gel, Two-Dimensional; D005347:Fibroblasts / Q000032:analysis / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008564:Membrane Potentials / Q000187:drug effects; D009994:Osmolar Concentration; D010455:Peptides / Q000032:analysis; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis; D012964:Sodium / Q000032:analysis / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects
D000596:Amino Acids; D002241:Carbohydrates; D002412:Cations; D010455:Peptides; D011506:Proteins; D012964:Sodium