The biosynthesis of ribulose bisphosphate carboxylase. Uncoupling of the synthesis of the large and small subunits in isolated soybean leaf cells.
Isolated leaf cells from soybean (Glycine max) incorporate [35S]methionine into protein at a linear rate for at least 5h. Analysis of the products of incorporation by one-dimensional and two-dimensional polyacrylamide gel electrophoresis shows that major products are the large and small subunits of the chloroplast enzyme, ribulose bisphosphate carboxylase. The large subunit is synthesized by chloroplast ribosomes and the small subunit by cytoplasmic ribosomes. Addition of chloramphenicol to the cells reduces incorporation into the large subunit without affecting incorporation into the products of cytoplasmic ribosomes. Addition of cycloheximide or 2-(4-methyl-2,6-dinitroanilino)-N-methylpropionamide stops incorporation into the small subunit, but large subunit continues to be made for at least 4 h.  For accurate estimates of incorporation into the large subunit, it is essential to use two-dimensional gel electrophoresis, because the large subunit region on one-dimensional gels is contaminated with the products of cytoplasmic ribosomes. Newly synthesized large subunits continue to enter complete molecules of ribulose bisphosphate carboxylase in the absence of small subunit synthesis. These results suggest that, in contrast to the situation in algal cells, the synthesis of the two subunits of ribulose bisphosphate carboxylase in the different subcellular compartments of higher plant cells is not tightly coupled over short time periods, and that a pool of small subunits exists in these cells. The results are disucssed in relation to possible mechanisms for the integration of the synthesis of the large and small subunits of ribulose bisphosphate carboxylase.
D002262:Carboxy-Lyases / Q000096:biosynthesis*; D002701:Chloramphenicol / Q000494:pharmacology; D002736:Chloroplasts / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D003624:Darkness; D007700:Kinetics; D046911:Macromolecular Substances; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D010455:Peptides / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D012273:Ribulose-Bisphosphate Carboxylase / Q000096:biosynthesis*; D013025:Glycine max
D046911:Macromolecular Substances; D010455:Peptides; D002701:Chloramphenicol; D003513:Cycloheximide; D008715:Methionine; D002262:Carboxy-Lyases; D012273:Ribulose-Bisphosphate Carboxylase