Commentary on 'Intermediate Reactions in Protein Biosynthesis'.
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to M B Hoagland.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
The aminoacylation of tRNA has been investigated in relation to protein aynthesis in living HeLa cells. In cells growing normally, the rates of tRNA charing are compatible with the observed entry of amino acids into protein. In contrast, when protein synthesis is inhibited 95--98% by either reduced temperature or cycloheximide, aminoacylation of tRNA is relatively unaffected. We conclude that, under these conditions, the aminoacylation of tRNA is uncoupled from subsequent steps in protein synthesis. These results provide for the first time a possible biological role for the observed aminoacyl-tRNA hydrolase activities of the tRNA synthetases.
A simple reaction system was developed to examine the binding of polysomes to membranes of the endoplasmic reticulum and to investigate the fate of ribosomes and nascent chains during protein synthesis in vitro. The system conssited of Sephadex G-25 treated post-mitochondrial fraction prepared from rat liver (Sephadex-PM) as a source of membranes, and radioactive free polysomes prepared from another rat liver. The following results were obtained. 1. Nascent chains on free polysomes labeled in vivo were transferred to membranes in vitro. The process required protein synthesis. 2. This reaction occurred in two steps: a) Binding of the free polysomes to membranes in the absence of protein synthesis. b) Release of ribosomes, leaving nascent chains on the membranes, requiring protein syntehsis. 3. A portion of the ribosomes found on membranes in vivi (membrane-bound ribosomes) was also released from the membranes during incubation in vitro, leaving their nascent chains on the membranes. The significance of the transfer of nascent chains from free polysomes to membranes in vitro is discussed in the light of known polysome-membrane interaction in vivo.