Relaxation kinetics of E coli ribosomes: evidence for the reaction of 30S . IF3 complex with 50S ribosomal subunits.
Addition of initiation factor IF3 to solutions of E.coli ribosomes dramatically alters their behavior in pressure-jump relaxation kinetic experiments in which 90 degrees light-scattering is used to monitor the macromolecular reaction. The effect of IF3 on relaxation processes attributed to "tight" couples is strongly dependent on the Mg2+ concentration. At 2.5 mM Mg2+, addition of 1 molar equivalent of IF3 decreases the relaxation amplitude by a factor of 3 relative to ribosome solutions without IF3. However, at 5.0 mM Mg2+, addition of 1 molar equivalent of IF3 produces a marked increase in the relaxation amplitude, by a factor of 2-8 fold relative to ribosomes in the absence of IF3. IF3 has no effect on the relaxation process attributed to "loose" couples at 10 mM Mg2+. While we are unable to propose a precise mechanism for IF3 action with the data on hand, our results require that the 30S . IF3 complex either reacts with the 50S subunit, forming a 70S . IF3 intermediate, or acts as a pool of reactive 30S subunit. Further kinetic evidence is required to distinguish between these possible pathways.
Chaires
J B
JB
Biochemistry and Biophysics Section, Biological Sciences Group, University of Connecticut Storrs, CT 06268 USA.
Kegeles
G
G
Wahba
A J
AJ