Localization of two domains of a mutant form of Escherichia coli protein L7/L12 that binds the large ribosomal subunit as a single dimer.
Escherichia coli ribosomal protein L7/L12 occurs on the large subunit as two dimers: one dimer is extended and comprises the stalk, while the second dimer is folded and occupies a site on the subunit body. A variant protein, in which all 18 amino acids of the flexible hinge region that links separate N-terminal and C-terminal domains of L7/L12 has been deleted, binds the subunit as a single dimer and does not generate stalks that are visible in electron micrographs. Monoclonal antibodies directed against each domain of the protein have been used to localize the variant in electron micrographs of 50S subunits. Both C-terminal domains are seen at a shoulder of the subunit, near its edge as viewed in the most common quasisymmetric projection. N-terminal domains are placed on the subunit body, about 50 A from the C-terminal domains. The antibody to the N-terminal domain also causes dissociation of the variant dimer from the particle and the formation of oligomeric antibody-protein dimer complexes. Similar complexes were seen previously (Olson HM et al (1986) J Biol Chem 261, 6924-6936) when this antibody induced dissociation of one dimer of the native protein. We conclude that the shortened variant most probably occupies the lower-affinity site on the subunit that is normally filled by the stalk dimer.
Wiggers
R J
RJ
Department of Biological Chemistry, UCLA School of Medicine, University of California, Los Angeles 90095-1737, USA.
Hadian
H
H
Traut
R R
RR
Oleinikov
A V
AV
Glitz
D G
DG
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D019281:Dimerization; D004926:Escherichia coli / Q000235:genetics* / Q000648:ultrastructure; D009154:Mutation; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary; D012269:Ribosomal Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D012269:Ribosomal Proteins; C016700:ribosomal protein L7-L12