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. Author manuscript; available in PMC: 2017 Jan 21.
Published in final edited form as: Nature. 2016 Jul 21;535(7612):444–447. doi: 10.1038/nature18622

Figure 1. De novo transcription initiation by ATP and NCINs.

Figure 1

a. Structures of ATP, NAD+, NADH, and dpCoA. Red, identical atoms. b. Initial RNA products of in vitro transcription reactions with ATP, NAD+, NADH, or dpCoA as initiating nucleotide and [α32P]-CTP as extending nucleotide (E. coli RNAP; PgadY). Products were treated with RppH (processes 5′-triphosphate RNA to 5′-monophosphate RNA and 5′-NTP to 5′-NDP/5′-NMP9,14) or NudC (processes 5′-NAD+/NADH-capped RNA to 5′-monophosphate RNA6) as indicated. c, d. Full-length RNA products of in vitro transcription reactions with ATP, NAD+, NADH, or dpCoA as initiating nucleotide and [α32P]-CTP, GTP, and UTP as extending nucleotides (c), or with [γ32P]-ATP or [α32P]-NAD+ as initiating nucleotide and CTP, GTP, and UTP as extending nucleotides (d) (E. coli RNAP; PrnaI fused to an A-less cassette ). M, 100-nt marker. For gel source data, see Supplementary Figure 1.