Effect of amino acid deprivation on initiation of protein synthesis in rat hepatocytes

W. V. Everson, K. E. Flaim, D. M. Susco, S. R. Kimball, L. S. Jefferson

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Abstract

Conditions were defined for maintaining optimal protein synthetic activity in suspensions of freshly isolated rat hepatocytes. Under these conditions, isolated hepatocytes exhibited rates of protein synthesis and levels of polysomal aggregation equivalent to those observed in vivo and in perfused liver. Deprivation of total amino acids or single, essential amino acids resulted in a rapid decrease in the rate of protein synthesis, which was readily reversed by readdition of the deficient amino acid(s). The decrease was accompanied by a disaggregation of polysomes and an inhibition of 43S initiation complex formation, which was indicative of a limitation in the rate of initiation of protein synthesis. Extracts prepared from perfused liver deprived of amino acids were inhibitory to initiation of protein synthesis in reticulocyte lysate. The inhibition in reticulocyte lysate was accompanied by an increase in phosphorylation of the α-subunit of eukaryotic initiation factor 2 (eIF-2), suggesting activation of an eIF-2α kinase or inhibition of a phosphatase in amino acid-deprived hepatocytes. This suggestion was confirmed by prelabeling hepatocytes with 32P(i) before amino acid deprivation. Incorporation of 32P(i) into eIF-2α was two- to threefold higher in lysine-deprived cells than in hepatocytes incubated in fully supplemented medium. Overall, the results indicated that an increase in eIF-2α phosphorylation was responsible for the defect in initiation of protein synthesis caused by amino acid deprivation.

Original languageEnglish (US)
Pages (from-to)25/1
JournalAmerican Journal of Physiology - Cell Physiology
Volume256
Issue number1
StatePublished - 1989

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cell Biology

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