DARG_THEA5
ID DARG_THEA5 Reviewed; 360 AA.
AC P0DV57;
DT 03-AUG-2022, integrated into UniProtKB/Swiss-Prot.
DT 03-AUG-2022, sequence version 1.
DT 03-AUG-2022, entry version 1.
DE RecName: Full=DNA ADP-ribosyl glycohydrolase {ECO:0000303|PubMed:27939941};
DE Short=DarG {ECO:0000303|PubMed:27939941};
DE EC=3.2.2.- {ECO:0000269|PubMed:27939941};
DE AltName: Full=Antitoxin DarG {ECO:0000303|PubMed:27939941};
GN Name=darG {ECO:0000303|PubMed:27939941};
GN ORFNames=TO73_1569 {ECO:0000312|EMBL:ALJ91410.1};
OS Thermus aquaticus (strain ATCC BAA-2747 / Y51MC23).
OC Bacteria; Deinococcus-Thermus; Deinococci; Thermales; Thermaceae; Thermus.
OX NCBI_TaxID=498848;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=BAA-2747 / Y51MC23;
RX PubMed=26465632; DOI=10.1371/journal.pone.0138674;
RA Brumm P.J., Monsma S., Keough B., Jasinovica S., Ferguson E.,
RA Schoenfeld T., Lodes M., Mead D.A.;
RT "Complete Genome Sequence of Thermus aquaticus Y51MC23.";
RL PLoS ONE 10:e0138674-e0138674(2015).
RN [2]
RP FUNCTION AS AN ANTITOXIN, AND EXPRESSION IN M.TUBERCULOSIS.
RX PubMed=32634279; DOI=10.1111/mmi.14571;
RA Zaveri A., Wang R., Botella L., Sharma R., Zhu L., Wallach J.B., Song N.,
RA Jansen R.S., Rhee K.Y., Ehrt S., Schnappinger D.;
RT "Depletion of the DarG antitoxin in Mycobacterium tuberculosis triggers the
RT DNA-damage response and leads to cell death.";
RL Mol. Microbiol. 114:641-652(2020).
RN [3]
RP FUNCTION AS AN ANTITOXIN, AND MUTAGENESIS OF ASN-22 AND LYS-80.
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).
RN [4] {ECO:0007744|PDB:5M31, ECO:0007744|PDB:5M3E}
RP X-RAY CRYSTALLOGRAPHY (1.67 ANGSTROMS) OF 1-155 WITH AND WITHOUT ADPR,
RP FUNCTION AS AN ANTITOXIN, FUNCTION AS AN ADP-RIBOSYL GLYCOHYDROLASE,
RP CATALYTIC ACTIVITY, EXPRESSION IN E.COLI, SUBUNIT, DOMAIN, POSSIBLE ACTIVE
RP SITE, AND MUTAGENESIS OF ASN-22; LYS-29; LYS-80; HIS-82; TRP-83 AND
RP GLY-119.
RX PubMed=27939941; DOI=10.1016/j.molcel.2016.11.014;
RA Jankevicius G., Ariza A., Ahel M., Ahel I.;
RT "The Toxin-Antitoxin System DarTG Catalyzes Reversible ADP-Ribosylation of
RT DNA.";
RL Mol. Cell 64:1109-1116(2016).
CC -!- FUNCTION: Antitoxin component of a hybrid type II/IV toxin-antitoxin
CC (TA) system. De-ADP-ribosylates DNA modified on thymidine by its
CC cognate toxin DarT, which neutralizes the activity of cognate toxin
CC DarT. Upon expression in E.coli neutralizes the effect of cognate toxin
CC DarT (PubMed:27939941). Upon expression in M.tuberculosis neutralizes
CC the toxic effects of endogenous DarT (PubMed:32634279).
CC {ECO:0000269|PubMed:27939941, ECO:0000269|PubMed:32634279}.
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:27939941};
CC -!- SUBUNIT: Probably forms a complex with cognate toxin DarT; this complex
CC might decrease ADP-ribosylation activity of DarT.
CC {ECO:0000305|PubMed:27939941}.
CC -!- DOMAIN: The macro domain alone is able to de-ADP-ribosylate substrate
CC in vitro and is almost as effective at neutralizing DarT in growth
CC experiments as full-length protein. {ECO:0000269|PubMed:27939941}.
CC -!- SIMILARITY: Belongs to the DarG ADP-ribosyl glycohydrolase family.
CC {ECO:0000305}.
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DR EMBL; CP010822; ALJ91410.1; -; Genomic_DNA.
DR PDB; 5M31; X-ray; 1.67 A; A=1-155.
DR PDB; 5M3E; X-ray; 2.50 A; A=1-155.
DR PDBsum; 5M31; -.
DR PDBsum; 5M3E; -.
DR PROSITE; PS51154; MACRO; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Toxin-antitoxin system.
FT CHAIN 1..360
FT /note="DNA ADP-ribosyl glycohydrolase"
FT /id="PRO_0000456047"
FT DOMAIN 1..155
FT /note="Macro"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00490"
FT ACT_SITE 80
FT /note="Nucleophile"
FT /evidence="ECO:0000305|PubMed:27939941"
FT BINDING 8..9
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000269|PubMed:27939941,
FT ECO:0007744|PDB:5M3E"
FT BINDING 20..22
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000269|PubMed:27939941,
FT ECO:0007744|PDB:5M3E"
FT BINDING 31..34
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000269|PubMed:27939941,
FT ECO:0007744|PDB:5M3E"
FT BINDING 79
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000269|PubMed:27939941,
FT ECO:0007744|PDB:5M3E"
FT BINDING 117..121
FT /ligand="ADP-D-ribose"
FT /ligand_id="ChEBI:CHEBI:57967"
FT /evidence="ECO:0000269|PubMed:27939941,
FT ECO:0007744|PDB:5M3E"
FT MUTAGEN 22
FT /note="N->A: Loss of de-ADP-ribosylation (of macro domain
FT fragment). Macro domain does not neutralize DarT in E.coli,
FT nor have enzyme activity in vitro; when associated with A-
FT 80."
FT /evidence="ECO:0000269|PubMed:27939941,
FT ECO:0000269|PubMed:34408320"
FT MUTAGEN 29
FT /note="K->E: Reduced de-ADP-ribosylation (of macro domain
FT fragment)."
FT /evidence="ECO:0000269|PubMed:27939941"
FT MUTAGEN 80
FT /note="K->A: Loss of de-ADP-ribosylation (of macro domain
FT fragment), whole protein does not neutralize DarT in growth
FT experiments, does decrease activity of DarT slightly in
FT vitro. Does not neutralize M.tuberculosis DarT in vivo.
FT Macro domain does not neutralize DarT in E.coli, nor have
FT enzyme activity in vitro; when associated with A-22."
FT /evidence="ECO:0000269|PubMed:27939941,
FT ECO:0000269|PubMed:32634279, ECO:0000269|PubMed:34408320"
FT MUTAGEN 82
FT /note="H->A: Slower than wild-type de-ADP-ribosylation (of
FT macro domain fragment)."
FT /evidence="ECO:0000269|PubMed:27939941"
FT MUTAGEN 83
FT /note="W->A: Slower than wild-type de-ADP-ribosylation (of
FT macro domain fragment)."
FT /evidence="ECO:0000269|PubMed:27939941"
FT MUTAGEN 119
FT /note="G->E: Greatly reduced de-ADP-ribosylation (of macro
FT domain fragment)."
FT /evidence="ECO:0000269|PubMed:27939941"
SQ SEQUENCE 360 AA; 40531 MW; 8DCE2B38EEFF88AB CRC64;
MLRFVRGNLL EAPVEALVNT VNTVGVMGKG VALQFKRAFP DNYQAYVKAC ERGQVQIGRI
FVYDRGPLAQ PRYIFNFPTK KHWRHPSRME YVEEGLKDLV CRIQELRVRS IALPPLGAGN
GGLPWPEVKQ RIQEALEALE GVEVWVYEPV ENPKAHSIVP LKTKPRLTPA RAALLKLFGL
YGALGEPLGR LEAQKLAYFL QEAGLDLKLD FACKQFGPYA EPLNHVLARL EGHYIQGFGD
RTGISQIRLK PQALDEAVLF LADYPKADEA ATRAADWVKG FETPYGLELL ATVHWAVRHE
GARDWASLQK RLQAWNPRKA TFPKTHLQVA LDALLKRGAL RPEEWQDRPP KLPANVAQEA
