DARG_ECO27
ID DARG_ECO27 Reviewed; 355 AA.
AC B7UP19;
DT 03-AUG-2022, integrated into UniProtKB/Swiss-Prot.
DT 03-AUG-2022, sequence version 2.
DT 03-AUG-2022, entry version 44.
DE RecName: Full=DNA ADP-ribosyl glycohydrolase {ECO:0000303|PubMed:32023456};
DE Short=DarG {ECO:0000303|PubMed:32023456};
DE EC=3.2.2.- {ECO:0000269|PubMed:32023456};
DE AltName: Full=Antitoxin DarG {ECO:0000303|PubMed:32023456};
GN Name=darG {ECO:0000303|PubMed:32023456};
GN OrderedLocusNames=E2348C_1092 {ECO:0000312|EMBL:CAS08640.1};
OS Escherichia coli O127:H6 (strain E2348/69 / EPEC).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=574521;
RN [1] {ECO:0000312|EMBL:CAS08640.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=E2348/69 / EPEC;
RX PubMed=18952797; DOI=10.1128/jb.01238-08;
RA Iguchi A., Thomson N.R., Ogura Y., Saunders D., Ooka T., Henderson I.R.,
RA Harris D., Asadulghani M., Kurokawa K., Dean P., Kenny B., Quail M.A.,
RA Thurston S., Dougan G., Hayashi T., Parkhill J., Frankel G.;
RT "Complete genome sequence and comparative genome analysis of
RT enteropathogenic Escherichia coli O127:H6 strain E2348/69.";
RL J. Bacteriol. 191:347-354(2009).
RN [2]
RP FUNCTION AS AN ANTITOXIN, FUNCTION AS AN ADP-RIBOSYL GLYCOHYDROLASE,
RP SUBUNIT, DOMAIN, AND MUTAGENESIS OF LYS-80.
RC STRAIN=E2348/69 / EPEC;
RX PubMed=32023456; DOI=10.1016/j.celrep.2020.01.014;
RA Lawaree E., Jankevicius G., Cooper C., Ahel I., Uphoff S., Tang C.M.;
RT "DNA ADP-Ribosylation Stalls Replication and Is Reversed by RecF-Mediated
RT Homologous Recombination and Nucleotide Excision Repair.";
RL Cell Rep. 30:1373-1384(2020).
RN [3]
RP FUNCTION AS AN ANTITOXIN, FUNCTION AS AN ADP-RIBOSYL GLYCOHYDROLASE, AND
RP DOMAIN.
RC STRAIN=E2348/69 / EPEC;
RX PubMed=34408320; DOI=10.1038/s41586-021-03825-4;
RA Schuller M., Butler R.E., Ariza A., Tromans-Coia C., Jankevicius G.,
RA Claridge T.D.W., Kendall S.L., Goh S., Stewart G.R., Ahel I.;
RT "Molecular basis for DarT ADP-ribosylation of a DNA base.";
RL Nature 596:597-602(2021).
CC -!- FUNCTION: Antitoxin component of a hybrid type II/IV toxin-antitoxin
CC (TA) system. De-ADP-ribosylates DNA modified by its cognate toxin DarT,
CC which neutralizes the activity of cognate toxin DarT. Neutralizes DarT
CC in vivo in 2 ways, by reversing the ADP-ribosylation via the DarG macro
CC domain and by interacting with (and presumably sequestering) DarT via
CC the DarG C-terminal domain (PubMed:32023456). In vivo the macrodomain
CC alone neutralizes DarT toxin from non-cognate T.aquaticus. The same
CC domain de-ADP-ribosylates gDNA (PubMed:34408320).
CC {ECO:0000269|PubMed:32023456, ECO:0000269|PubMed:34408320}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an N-(ADP-alpha-D-ribosyl)-thymidine in DNA + H2O = a
CC thymidine in DNA + ADP-D-ribose; Xref=Rhea:RHEA:71655, Rhea:RHEA-
CC COMP:13556, Rhea:RHEA-COMP:18051, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:57967, ChEBI:CHEBI:137386, ChEBI:CHEBI:191199;
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:71656;
CC Evidence={ECO:0000269|PubMed:32023456, ECO:0000269|PubMed:34408320};
CC -!- SUBUNIT: Interacts with DarT (shown using mutant G49E), primarily via
CC the C-terminus of DarG, which probably sequesters DarT and inhibits it.
CC {ECO:0000269|PubMed:32023456}.
CC -!- DOMAIN: The macro domain (residues 1-16 joined to 112-330) alone is
CC able to de-ADP-ribosylate substrate in vitro and is almost as effective
CC at neutralizing DarT in growth experiments as full-length protein. The
CC C-terminal domain has no de-ADP-ribosylation activity, but is able to
CC neutralize DarT mutant G49D and interacts with DarT mutant G49D.
CC {ECO:0000269|PubMed:32023456, ECO:0000269|PubMed:34408320}.
CC -!- SIMILARITY: Belongs to the DarG ADP-ribosyl glycohydrolase family.
CC {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CAS08640.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305};
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DR EMBL; FM180568; CAS08640.1; ALT_INIT; Genomic_DNA.
DR EnsemblBacteria; CAS08640; CAS08640; E2348C_1092.
DR KEGG; ecg:E2348C_1092; -.
DR HOGENOM; CLU_049707_0_0_6; -.
DR OMA; WIVNFPT; -.
DR Proteomes; UP000008205; Chromosome.
DR Gene3D; 3.40.220.10; -; 1.
DR InterPro; IPR002589; Macro_dom.
DR InterPro; IPR043472; Macro_dom-like.
DR Pfam; PF01661; Macro; 1.
DR SMART; SM00506; A1pp; 1.
DR SUPFAM; SSF52949; SSF52949; 1.
DR PROSITE; PS51154; MACRO; 1.
PE 1: Evidence at protein level;
KW Hydrolase; Toxin-antitoxin system.
FT CHAIN 1..355
FT /note="DNA ADP-ribosyl glycohydrolase"
FT /id="PRO_0000456044"
FT DOMAIN 14..155
FT /note="Macro"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00490"
FT REGION 125..343
FT /note="C-terminal domain; neutralizes DarT mutant G49D in
FT vivo, interacts with DarT mutant G49E"
FT /evidence="ECO:0000269|PubMed:32023456"
FT ACT_SITE 80
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:P0DV57"
FT BINDING 13..15
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:P0DV57"
FT BINDING 20..22
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:P0DV57"
FT BINDING 31..34
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:P0DV57"
FT BINDING 79
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:P0DV57"
FT BINDING 117..121
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000250|UniProtKB:P0DV57"
FT MUTAGEN 80
FT /note="K->A: Loss of de-ADP-ribosylation (in full-length
FT and macro domain fragment), mutation in macro fragment no
FT longer neutralizes DarT mutant G49D, mutation in full-
FT length protein does neutralize DarT G49D toxicity in vivo."
FT /evidence="ECO:0000269|PubMed:32023456"
SQ SEQUENCE 355 AA; 40673 MW; CE0963F39580308F CRC64;
MITYTQGNLL DAPVEALVNT VNTVGVMGKG IALMFKERFP ENMKVYALAC KQKQVITGKM
FITETGELMG PRWIVNFPTK QHWRADSRME WIEDGLQDLR RFLIEENVQS IAIPPLGAGN
GGLNWPDVRA QIESALGDLQ DVDILIYQPT EKYQNVAKST GVKKLTPARA AIAELVRRYW
VLGMECSLLE IQKLAWLLQR AIEQHQQDDI LKLRFEAHYY GPYAPNLNHL LNALDGTYLK
AEKRIPDSQP LDVIWFNDQK KEHVNAYLNN EAREWLPALE QVSQLIDGFE SPFGLELLAT
VDWLLSRGEC QPTLDSVKEG LHQWPAGERW ASRKLRLFDN NNLQFAINRV MEFHC
