It has been more than 50 years since the discovery of dinucleoside polyphosphates (Np_nNs) and yet their roles and mechanisms of action remain unclear. Here, we show that both methylated and non-methylated Np_nNs serve as RNA caps in Escherichia coli. Np_nNs are excellent substrates for T7 and E. coli RNA polymerases (RNAPs) and efficiently initiate transcription. We demonstrate, that the E. coli enzymes RNA 5′-pyrophosphohydrolase (RppH) and bis(5′-nucleosyl)-tetraphosphatase (ApaH) are able to remove the Np_nN-caps from RNA. ApaH is able to cleave all Np_nN-caps, while RppH is unable to cleave the methylated forms suggesting that the methylation adds an additional layer to RNA stability regulation. Our work introduces a different perspective on the chemical structure of RNA in prokaryotes and on the role of RNA caps. We bring evidence that small molecules, such as Np_nNs are incorporated into RNA and may thus influence the cellular metabolism and RNA turnover.

Despite substantial evidence of short Au⋅⋅⋅H−X contacts derived from a number of X‐ray structures of Au^I compounds, the nature of Au^I⋅⋅⋅H bonding in these systems has not been clearly understood. Herein, we present the first spectroscopic evidence for an intramolecular Au^I⋅⋅⋅H⁺−N hydrogen bond in a [Cl−Au−L]⁺ complex, where L is a protonated N‐heterocyclic carbene. The complex was isolated in the gas phase and characterized with helium‐tagging infrared photodissociation (IRPD) spectra, in which H⁺−N‐mode‐derived bands evidence the intramolecular Au^I⋅⋅⋅H⁺−N bond. Quantum chemical calculations reproduce the experimental IRPD spectra and allow to characterize the intramolecular Au⋅⋅⋅H⁺−N bonding with a short r_Au⋅⋅⋅H distance of 2.17 Å and an interaction energy of approximately −10 kcal mol⁻¹. Various theoretical descriptors of chemical bonding calculated for the Au⋅⋅⋅H⁺−N interaction provide strong evidence for a hydrogen bond of moderate strength.