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Nature of eukaryotic proteins required for joining of 40S and 60S ribosomal subunits
Authors:Cesar Nombela  Nohelly A. Nombela  Severo Ochoa  William C. Merrick  W. French Anderson
Affiliation:Roche Institute of Molecular Biology, Department of Biochemistry Nutley, New Jersey 07110 USA;Molecular Hematology Branch, National Heart and Lung Institute National Institutes of Health, Bethesda, Maryland 20014 USA
Abstract:A mixture of 40S and 60S subunits from salt-washed rabbit reticulocyte ribosomes fails to promote methionyl-puromycin synthesis under conditions in which an AUG-40S-Met-tRNAi initiation complex, but not an 80S complex, is readily formed. This suggests that the inability of the system to form methionyl-puromycin is due to failure of the subunits to join. When Artemia salina 60S subunits are substituted for their reticulocyte counterparts, the resulting hybrid system readily forms an 80S initiation complex and synthesizes methionyl-puromycin. Activity of the reticulocyte 60S subunits can be restored by factors IF-M2A and IF-M2B. This suggests that one or both of these factors may be 60S proteins, essential for subunit joining, that may be removed from ribosomes by salt washing procedures.
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