RNA polymerase: linear competitive inhibition by bis-(3′→5′)-cyclic dinnucleotides,NpNp

Abstract

We have investigated the possible role of the bis-(3' to 5')-cyclic dinucleotides UpUp and ApUp as kinetic inhibitors of the DNA dependent RNA polymerase enzyme of E. coli, using T7 delta D111 deletion mutant DNA and several synthetic DNA polymers as templates. We have established that UpUp is a linear competitive inhibitor of the initiation phase of the polymerization (Ki = 28 microM using T7 delta D111 DNA as a template), but that it has no effect when added during the elongation phase. The compound ApUp is an inhibitor of the reaction only when poly(dA-T).poly(dA-T) is used as a template, and UpUp is an inhibitor of the reaction when poly(dA).poly(dT) was employed as the DNA template.

Effect of low nucleotide concentrations on abortive elongation catalysed by wheat-germ RNA polymerase II

“…The importance of the 2'-hydroxy group for the binding of nucleotides (substrates or inhibitors) is also strengthened by the observation that ara-ATP and 2'-amino-ATP are not inhibitors of transcription by E. coli RNA polymerase, in contrast with xylo-ATP and 3'-amino-ATP (Dennis et al, 1980;Armstrong & Eckstein, 1976). Bis-(3'-l5')-cyclic dinucleotides also behave as very potent inhibitors of the initiation phase of the polymerization with E. coli RNA polymerase (Hsu & Dennis, 1982). Therefore it seems possible that 3'-dATP might, to some extent, mimic the role of the activator or discriminator nucleotide described for the prokaryotic RNA polymerase reactions (Shimamoto & Wu, 1980a,b) The results obtained with poly[d(A-A-T)] * poly-[d(T-T-A)] as template (Fig.…”

Cyclic di-GMP: the First 25 Years of a Universal Bacterial Second Messenger

“…This suggests that the half-life of pGpG may be prolonged selectively in vivo to allow priming of transcription. In this context, it is worth noting that cyclic dinucleotides have been shown to inhibit the activity of RNA polymerase in vitro under certain conditions (391).…”

Quantitative determination of cyclic diguanosine monophosphate concentrations in nucleotide extracts of bacteria by matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry

“…C-di-GMP and c-di-AMP used in the construction of the standard curve were chemically synthesized as described [10][11]. The MS-spectrum of c-di-GMP has distinct peaks at a mass to charge ratio of 689, 711 and 727 Da, which corresponds…”