A mutant elongation factor Tu which does not immobilize the ribosome upon binding of kirromycin.

In the accompanying paper we have shown that polypeptide synthesis sustained by the mutant elongation factor EF-TuBO is inhibited by kirromycin. Here we have searched for the primary site of inhibition in the elongation cycle. It is demonstrated that in the presence of the antibiotic EF-TuBO can form a complex with aminoacyl-tRNA and GTP and that the complex is able to bind to ribosomes programmed with poly(U). Like its wild-type counterpart, EF-TuBO . GDP can form a quaternary complex with aminoacyl-tRNA and kirromycin but, unlike the wild-type quaternary complex, the mutant complex fails to associate with the ribosome. This explains the recessive nature of the tuf B mutation in cells producing kirromycin-resistant EF-TuA and EF-TuBO. It also suggests a mechanism for the inhibition by kirromycin of EF-TuBO-dependent polypeptide synthesis.