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. 2020 Oct 23;11:5388. doi: 10.1038/s41467-020-19166-1

Fig. 1. Proteome-wide DRaCALA screen identifies both conserved categories of binding targets and novel targets.

Fig. 1

a The Bacillus anthracis ORF donor vector library was recombined by Gateway cloning into overexpression vectors to generate ORFs with an N-terminal His-tag or HisMBP tag. The plasmids were transformed into E. coli for overexpression of recombinant proteins. Lysates of each ORF overexpressed in E. coli were assayed for binding to pppGpp, ppGpp, and pGpp using DRaCALA. b List of identified (p)ppGpp-binding targets in E. coli, S. aureus, and B. anthracis and pGpp-binding targets in B. anthracis. pppGpp results were obtained using both His-tagged and His-MBP-tagged proteins. ppGpp and pGpp results were obtained using His-MBP-tagged proteins. NT not tested, NA not available due to the lack of homologous gene. c, d Schematics of pathways differentially regulated by pGpp and (p)ppGpp: c Enzymes in purine nucleotide synthesis, including HprT, Xpt, Gmk, and GuaC, bound both pGpp and (p)ppGpp; d GTPases, involved in ribosome biogenesis and translational control (Obg, HflX, Der, RbgA, and Era), bound (p)ppGpp, but not pGpp.