Control of protein synthesis by hemin. Isolation and characterization of a supernatant factor from rabbit reticulocyte lysate

Abstract

The regulation of protein synthesis by hemin in rabbit reticulocyte lysates is mediated by a hemin-controlled translational repressor protein (HCR) that inhibits polypeptide chain initiation. The effect of this translational inhibitor can be reversed by a high molecular weight protein in the post-ribosomal supernatant fraction. This supernatant factor has been purified approx. 700-fold. It is as effective in reversing the inhibition of protein synthesis due to an early form of HCR (intermediate HCR) as it is in stimulating protein synthesis in the absence of hemin. It is progressively less effective at reversing the inhibition of protein synthesis due to a late form of HCR (irreversible HCR), double-stranded RNA, and oxidized glutathione. The supernatant factor is chromatographically different from the initiation factor IF-MP, isolated from reticulocyte ribosomes, that can also overcome the inhibitory effect of HCR. The supernatant factor does not require hemin for activity, and its action is somewhat suppressed by a level of hemin that is optimal for protein synthesis.