Studies on stringent control in a cell-free system. Regulation by guanosine-5'-diphosphate-3'-diphosphate of the synthesis of elongation factor Tu.:Regulation by Guanosine-5'-Diphosphate-3'-Diphosphate of the Synthesis of Elongation Factor Tu

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The biosynthesis of elongation factor Tu (EF-Tu) has been studied in a cell-free system with DNA of the transducing phage λrifd18 as a template. It was found that the synthesis of EF-Tu in this system was inhibited by about 60% in the presence of 0.3 to 0.6mM guanosine-5'-diphosphate-3'-diphosphate (ppGpp). The syntheses of several ribosomal proteins encoded in this template, i.e. Ll, L10, L11, and L7/L12, were also depressed, whereas those of phage lambda proteins were rather enhanced by the addition of ppGpp.<br> By separating the reaction into two steps, i.e., transcription and translation, the effect of ppGpp was shown to occur at the level of transcription. Several analogs, such as guanosine-5'-diphosphate-3'-monophosphate (ppGp) and guanosine-5'-diphosphate (ppG), were without effect. The formation of mRNA for EF-Tu was assessed directly by specific hybridization with pTUA1 DNA carrying tufA gene. The results clearly indicated that the synthesis of tufB mRNA was severely and selectively inhibited by ppGpp.

Journal

  • J Biochem (Tokyo)

    J Biochem (Tokyo) 86 (2), 403-411, 1979

    The Japanese Biochemical Society

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