The β-D-manno-heptoses are immune agonists across kingdoms

Bacterial small molecule metabolites such as adenosine-diphosphate-D-glycero-β-D-manno-heptose (ADP-heptose) and their derivatives act as effective innate immune agonists in mammals. We showed that functional nucleotide-diphosphate-heptose biosynthetic enzymes (HBEs) are distributed widely in bacteria, archaea, eukaryotes, and viruses. We identified a conserved STTR5 motif as a hallmark of heptose nucleotidyltransferases that can synthesize not only ADP-heptose but also cytidine-diphosphate (CDP)- and uridine-diphosphate (UDP)-heptose. Both CDP- and UDP-heptoses are agonists that trigger stronger alpha-protein kinase 1 (ALPK1)-dependent immune responses than ADP-heptose in human and mouse cells and mice. We also produced ADP-heptose in archaea and verified its innate immune agonist functions. Hence, the β-D-manno-heptoses are cross-kingdom small molecule pathogen-associated molecular patterns that activate the ALPK1-dependent innate immune signaling cascade., , , Paper: **The β-D-*manno*-heptoses are immune agonists across kingdoms**

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**Descriptions**

This folder contains the enzymatic assay data of NDP-heptose biosynthetic enzymes (HBEs) data, phylogenesis of HBEs performing nucleotidyltransferase activities (HENases) beyond bacteria, source data of cytokines in mice, qPCR source data, and dual luciferase assay data.

**Enzymatic assay data of HBEs**

NDP-heptoses generated by HENases combined with appropriate HBEs in four-step reactions (Fig. 2G).

Synthesis of NDP-heptoses by different HBE combinations at a simulated physiological condition (pH 7.2, S7P 0.1 mM, ATP 3.5 mM, CTP 0.3 mM, and UTP 0.7 mM) (Fig. 3A).

**Phylogenesis**

Three .fasta files and three .nwk files were contained.

The “Viral HENase aligments.fas”, “Archaeal HENase aligments.fas”, “Eukaryotic HENase aligments.fas” contain the aligned sequences of represen...