Subunit association defects in Escherichia coli ribosome mutants lacking proteins S20 and L11.
The subunit association capacity of 30S and 50S ribosomal subunits from Escherichia coli mutants lacking protein S20 or L11 as well as of 50S subunits depleted of L7/L12 was tested by sucrose gradient centrifugation and by a nitrocellulose filtration method based on the protection from hydrolysis with peptidyl-tRNA hydrolase of ribosome-bound AcPhe-tRNA. It was found that the subunits lacking either S20 or L11 display an altered association capacity, while the 50S subunits lacking L7/L12 have normal association behavior. The association of S20-lacking 30S subunits is quantitatively reduced, especially at low Mg2+ concentrations (5-12 mM), and produces loosely interacting particles which dissociate during sucrose gradient centrifugation. The association of L11-lacking 50S subunits is quantitatively near-normal at all Mg2+ concentrations and produces loosely associating particles only at low Mg2+ concentrations (5-8 mM); the mechanism of their association with 30S subunits, however, or the structure of the resulting 30S-50S couples is altered in such a way as to cause the ejection of an AcPhe-tRNA molecule pre-bound to the 30S subunits in response to poly(U).
Götz
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F
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin-Dahlem.
Fleischer
C
C
Pon
C L
CL
Gualerzi
C O
CO
D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D001665:Binding Sites / Q000187:drug effects; D002499:Centrifugation, Density Gradient; D004926:Escherichia coli / Q000032:analysis / Q000235:genetics*; D006868:Hydrolysis; D008274:Magnesium / Q000494:pharmacology; D009154:Mutation; D011485:Protein Binding; D012343:RNA, Transfer / Q000032:analysis; D012269:Ribosomal Proteins / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D012270:Ribosomes / Q000502:physiology
D001426:Bacterial Proteins; D012269:Ribosomal Proteins; C027624:ribosomal protein L11; C029611:ribosomal protein S20; C040469:ribosomal protein S30; C016700:ribosomal protein L7-L12; D012343:RNA, Transfer; D008274:Magnesium