Structure of the bifunctional Rel enzyme from B. subtilis

Structural Basis for Regulation of the Opposing (p)ppGpp Synthetase and Hydrolase within the Stringent Response Orchestrator Rel.

(p)ppGpp synthase,ATP:GTP 3'-pyrophosphotransferase,ppGpp synthase I

MANEQVLTAEQVIDKARSYLSDEHIAFVEKAYLYAEDAHREQYRKSGEPYIIHPIQVAGILVDLEMDPSTIAGGFLHDVV
EDTDVTLDDLKEAFSEEVAMLVDGVTKLGKIKYKSQEEQQAENHRKMFVAMAQDIRVILIKLADRLHNMRTLKHLPQEKQ
RRISNETLEIFAPLAHRLGISKIKWELEDTALRYLNPQQYYRIVNLMKKKRAERELYVDEVVNEVKKRVEEVNIKADFSG
RPKHIYSIYRKMVLQNKQFNEIYDLLAVRILVNSIKDCYAVLGIIHTCWKPMPGRFKDYIAMPKPNMYQSLHTTVIGPKG
DPLEVQIRTFEMHEIAEYGVAAHWAYKEGKAANEGATFEKKLSWFREILEFQNESTDAEEFMESLKIDLFSDMVYVFTPK
GDVIELPSGSVPIDFSYRIHSEIGNKTIGAKVNGKMVTLDHKLRTGDIVEILTSKHSYGPSQDWVKLAQTSQAKHKIRQF
FKKQRREENVEKGRELVEKEIKNLDFELKDVLTPENIQKVADKFNFSNEEDMYAAVGYNGITALQVANRLTEKERKHHHH
HH

Bacillus subtilis subsp. subtilis str. 168

1 M Lithium chloride, 0.1 M Bicine pH 9.0, 10% PEG6000 (w/v), final pH 9.0

GST pulldown of individual domains and mutagenesis to disrupt dimerization interface

bifunctional (p)ppGpp synthease/hydrolase, stringent response, ribosome interacting, SIGNALING PROTEIN

MANEQVLTAEQVIDKARSYLSDEHIAFVEKAYLYAEDAHREQYRKSGEPYIIHPIQVAGILVDLEMDPSTIAGGFLHDVV
EDTDVTLDDLKEAFSEEVAMLVDGVTKLGKIKYKSQEEQQAENHRKMFVAMAQDIRVILIKLADRLHNMRTLKHLPQEKQ
RRISNETLEIFAPLAHRLGISKIKWELEDTALRYLNPQQYYRIVNLMKKKRAERELYVDEVVNEVKKRVEEVNIKADFSG
RPKHIYSIYRKMVLQNKQFNEIYDLLAVRILVNSIKDCYAVLGIIHTCWKPMPGRFKDYIAMPKPNMYQSLHTTVIGPKG
DPLEVQIRTFEMHEIAEYGVAAHWAYKEGKAANEGATFEKKLSWFREILEFQNESTDAEEFMESLKIDLFSDMVYVFTPK
GDVIELPSGSVPIDFSYRIHSEIGNKTIGAKVNGKMVTLDHKLRTGDIVEILTSKHSYGPSQDWVKLAQTSQAKHKIRQF
FKKQRREENVEKGRELVEKEIKNLDFELKDVLTPENIQKVADKFNFSNEEDMYAAVGYNGITALQVANRLTEKERK