lithium-chloride and Chromosome-Deletion

Five-residue-long deletions centered on Ala63, Ala75, and Glu118 of ribosomal protein L7/L12 gave low mutant yields (5% or less) when the mutant genes were cloned in phage M13mp18 and controlled by the L10 promotor. Deletions of Glu118-Lys120 or Lys120 (the COOH-terminus of L7/L12) gave higher mutant yields, up to 50% with L7/L12 delta Lys120. L7/L12 delta Lys120 was not preferentially found in the S100 and not preferentially removed by LiCl washing, but was preferentially extracted from 70S ribosomes in the presence of 28-35% ethanol in 0.25-0.5 M NH4Cl. It follows that delta Lys120 destabilizes the ribosome-binding domain of ribosomal protein L7/L12 in an ethanol-containing solvent, which raises the question whether Lys120 is part of the ribosome-binding domain of L7/L12 during some step of protein synthesis or whether it is essential to preserve the conformation of the physiological ribosome-binding domain under structurally stressful conditions.

Topics: Amino Acid Sequence; Ammonium Chloride; Base Sequence; Binding Sites; Centrifugation, Density Gradient; Chlorides; Chromosome Deletion; Cloning, Molecular; Electrophoresis; Escherichia coli; Ethanol; Lithium; Lithium Chloride; Lysine; Molecular Sequence Data; Mutation; Ribosomal Proteins; Ribosomes