The mechanism of action of ppGpp on rRNA synthesis in vitro

Abstract

We have studied the mechanism of the specific inhibition of ribosomal RNA synthesis by ppGpp in a purified system using as templates E. coli DNA and DNA from lambdad5ilv, which carries a rRNA cistron from E. coli. Ribosomal RNA synthesis, as well as its inhibition by ppGpp, are critically salt-dependent. Of a number of guanosine phosphates tested, only pppGpp (MS II) mimicked the action of ppGpp, establishing the specificity of ppGpp. The two templates gave similar results for rRNA synthesis in all experiments. By using the initiation inhibitor rifampicin, we could show that the specific inhibition of rRNA synthesis by ppGpp is due to its effect on rRNA initiation. The somewhat variable inhibition of RNA synthesis in general by ppGpp is mainly or wholly a consequence of premature chain termination. We propose that ppGpp specifically inhibits rRNA synthesis by acting on the formation of the so-called "closed-promoter complex".