Globin mRNA translation on Artemia salina ribosomes with components from Friend leukemia cells.
Globin mRNA can be translated with relatively high efficiency in a fractionated cell-free system containing ribosomes prepared from cytst of Artemia salina. These ribosomes have unusually low endogenous activity for peptide synthesis in the absence of added mRNA. The system requires components from the postribosomal supernatant and from the 0.5 M KCl ribosomal wash fraction. Both these fractions were derived from either rabbit reticulocytes or unstimulated Friend leukemia cells that produce little or no hemoglobin. The activity of mRNA and enzyme fractions from rabbit reticulocytes and Friend leukemia cells were tested in this system in vitro for their ability to direct the synthesis of the alpha and beta chains of globin. The alpha:beta chain ratio synthesized from mRNA in the rabbit reticulocyte salt wash fraction was 4:1. The corresponding value for the 9-S mRNA fraction from the salt-washed reticulocyte ribosomes was 1:4, thus these two fractions appear to provide sources enriched in either alpha or beta globin mRNA. Under all conditions tested, the ratio and amounts of peptides formed in vitro appear to reflect mRNA composition. Globin mRNA from dimethysulfoxide-stimulated Friend leukemia cells when translated in vitro produced alpha and beta chains in a ratio of 1:1. These peptides are formed in the same ratio in the intact cells.
D000818:Animals; D002474:Cell-Free System; D002852:Chromatography, Ion Exchange; D033364:Decapoda / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005622:Friend murine leukemia virus; D005914:Globins / Q000096:biosynthesis*; D007700:Kinetics; D007942:Leukemia, Experimental / Q000378:metabolism*; D008099:Liver; D051379:Mice; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000097:blood*; D012343:RNA, Transfer / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D012333:RNA, Messenger; D005914:Globins; D012343:RNA, Transfer