Protein synthesis in the developing rat liver: participation of initiation factors eIF-2 and eIF-2B.
We studied the decline in protein synthesis in the developing liver in suckling rats (4 to 10 days) and adult rats (2 mo). The rate of protein synthesis was measured with a cell-free system and compared with the activity of two initiation factors, eukaryotic initiation factor-2 and eukaryotic initiation factor-2B, and with casein kinase II, which phosphorylates both factors in vitro. The specific activity of the three parameters decreased in adult rats compared with suckling rats and in parallel to the rate of protein synthesis. Quantification of eukaryotic initiation factor-2 in the ribosomal salt wash and in the postmicrosomal supernatant showed that both the specific activity and the levels of eukaryotic initiation factor-2, are much higher in the ribosomal salt wash fractions than in postmicrosomal supernatants, but no differences were found between the two age groups. The eukaryotic initiation factor-2/ribosome ratio was higher in adult rats than in suckling rats, and this parameter seems to be inversely proportional to the rate of protein synthesis. The phosphorylation state of eukaryotic initiation factor-2 alpha, as determined by isoelectric focusing followed by protein immunoblotting, revealed very low and equal levels of phosphorylation in the two animal groups. The lack of changes in eukaryotic initiation factor-2 levels and phosphorylation status suggests that a decrease in the activity or levels of eukaryotic initiation factor-2B could be responsible for the decrease in eukaryotic initiation factor-2 activity and account for, at least in part, the differences observed in the rates of protein synthesis.
García
A M
AM
Department of Investigation, Hospital Ramón y Cajal, Madrid, Spain.
Martín
M E
ME
Alcázar
A
A
Fando
J L
JL
Salinas
M
M
D000375:Aging / Q000378:metabolism; D000818:Animals; D000833:Animals, Suckling; D047390:Casein Kinase II; D002474:Cell-Free System / Q000378:metabolism; D015852:Eukaryotic Initiation Factor-2 / Q000502:physiology*; D005260:Female; D020662:Guanine Nucleotide Exchange Factors; D008099:Liver / Q000254:growth & development* / Q000378:metabolism*; D010766:Phosphorylation; D014176:Protein Biosynthesis; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011506:Proteins / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions / Q000378:metabolism; D014018:Tissue Distribution
D015852:Eukaryotic Initiation Factor-2; D020662:Guanine Nucleotide Exchange Factors; D011506:Proteins; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases