DART_MYCTU
ID DART_MYCTU Reviewed; 230 AA.
AC O53604; I6Y2M9; Q7DAJ1;
DT 03-AUG-2022, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-1998, sequence version 1.
DT 03-AUG-2022, entry version 108.
DE RecName: Full=DNA ADP-ribosyl transferase {ECO:0000303|PubMed:27939941};
DE Short=DarT {ECO:0000303|PubMed:27939941};
DE EC=2.4.2.- {ECO:0000269|PubMed:27939941};
DE AltName: Full=Toxin DarT {ECO:0000303|PubMed:27939941};
GN Name=darT {ECO:0000303|PubMed:32634279};
GN OrderedLocusNames=Rv0059 {ECO:0000312|EMBL:CCP42781.1};
OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae;
OC Mycobacterium; Mycobacterium tuberculosis complex.
OX NCBI_TaxID=83332;
RN [1] {ECO:0000312|EMBL:CCP42781.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=9634230; DOI=10.1038/31159;
RA Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., Harris D.E.,
RA Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K.,
RA Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K.,
RA Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K.,
RA Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J.,
RA Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S.,
RA Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S.,
RA Barrell B.G.;
RT "Deciphering the biology of Mycobacterium tuberculosis from the complete
RT genome sequence.";
RL Nature 393:537-544(1998).
RN [2] {ECO:0007744|PubMed:21969609}
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=21969609; DOI=10.1074/mcp.m111.011627;
RA Kelkar D.S., Kumar D., Kumar P., Balakrishnan L., Muthusamy B., Yadav A.K.,
RA Shrivastava P., Marimuthu A., Anand S., Sundaram H., Kingsbury R.,
RA Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., Katoch K., Katoch V.M.,
RA Kumar P., Chaerkady R., Ramachandran S., Dash D., Pandey A.;
RT "Proteogenomic analysis of Mycobacterium tuberculosis by high resolution
RT mass spectrometry.";
RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011).
RN [3]
RP FUNCTION AS A TOXIN, AND FUNCTION AS AN ADP-RIBOSYL TRANSFERASE.
RC STRAIN=H37Rv;
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).
RN [4]
RP FUNCTION AS A TOXIN, SUBUNIT, AND DISRUPTION PHENOTYPE.
RC STRAIN=H37Rv;
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 [5]
RP FUNCTION AS A TOXIN, FUNCTION AS AN ADP-RIBOSYL TRANSFERASE, OPERON
RP STRUCTURE, DOMAIN, AND DISRUPTION PHENOTYPE.
RC STRAIN=H37Rv;
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: Toxic component of a hybrid type II/IV toxin-antitoxin (TA)
CC system. Its toxic effect is neutralized by cognate antitoxin DarG
CC (PubMed:27939941). ADP-ribosylates ssDNA, preferentially in the motif
CC TTTW. Uncontrolled expression of DarT leads to ADP-ribosylation of the
CC origin of chromosomal replication DNA in cells (in vitro the most
CC heavily modified motifs are TTTT/A in the OriC lower strand) and growth
CC arrest (PubMed:34408320). Is very toxic to E.coli, it cannot be
CC expressed in E.coli (PubMed:27939941). Experiments in situ in which
CC antitoxin DarG levels are depleted (similar to overexpression of DarT)
CC lead to cell death; expression of wild-type DarG protein from
CC M.tuberculosis or T.aquaticus restores growth. Cells with decreased
CC levels of DarG are more sensitive to bedaquilline (targets
CC respiration), DNA-damaging drugs (mitomycin C, netropsin) and
CC transcription-targeted drugs (rifabutin and rifampicin). When DarG is
CC depleted, a DNA-damage response is induced and mutability is increased,
CC suggesting ADP-ribosylation of DNA is the toxic effect
CC (PubMed:32634279). {ECO:0000269|PubMed:27939941,
CC ECO:0000269|PubMed:32634279, ECO:0000269|PubMed:34408320}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a thymidine in DNA + NAD(+) = an N-(ADP-alpha-D-ribosyl)-
CC thymidine in DNA + H(+) + nicotinamide; Xref=Rhea:RHEA:71651,
CC Rhea:RHEA-COMP:13556, Rhea:RHEA-COMP:18051, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:17154, ChEBI:CHEBI:57540, ChEBI:CHEBI:137386,
CC ChEBI:CHEBI:191199;
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:71652;
CC Evidence={ECO:0000269|PubMed:27939941, ECO:0000269|PubMed:34408320};
CC -!- SUBUNIT: Forms a complex with cognate antitoxin DarG; this complex
CC neutralizes the toxic effect of DarT. {ECO:0000269|PubMed:32634279}.
CC -!- INDUCTION: Part of the dnaB-darT-darG operon.
CC {ECO:0000269|PubMed:34408320}.
CC -!- DOMAIN: The NAD(+)-binding element stabilizes the ADP-ribosylating
CC turn-turn (ARTT) loop which confers substrate specificity; both domains
CC contribute to ssDNA-binding. {ECO:0000250|UniProtKB:A0A0B0SG80}.
CC -!- DISRUPTION PHENOTYPE: A double darT-darG deletion shows no change in
CC growth in culture, upon infection of mice or upon exposure to a variety
CC of stresses (PubMed:32634279). Another group finds the double knockout
CC gives a competitive advantage over wild-type cells in liquid culture
CC growth experiments (PubMed:34408320). {ECO:0000269|PubMed:32634279,
CC ECO:0000269|PubMed:34408320}.
CC -!- SIMILARITY: Belongs to the DarT ADP-ribosyltransferase family.
CC {ECO:0000305}.
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DR EMBL; AL123456; CCP42781.1; -; Genomic_DNA.
DR RefSeq; NP_214573.1; NC_000962.3.
DR RefSeq; WP_003400548.1; NZ_NVQJ01000005.1.
DR SMR; O53604; -.
DR STRING; 83332.Rv0059; -.
DR PaxDb; O53604; -.
DR PRIDE; O53604; -.
DR GeneID; 45424018; -.
DR GeneID; 887006; -.
DR KEGG; mtu:Rv0059; -.
DR PATRIC; fig|83332.111.peg.66; -.
DR TubercuList; Rv0059; -.
DR eggNOG; COG4948; Bacteria.
DR OMA; HDYVPFY; -.
DR Proteomes; UP000001584; Chromosome.
DR InterPro; IPR029494; DarT.
DR Pfam; PF14487; DarT; 1.
PE 1: Evidence at protein level;
KW DNA-binding; Glycosyltransferase; Nucleotidyltransferase;
KW Reference proteome; Toxin-antitoxin system; Transferase.
FT CHAIN 1..230
FT /note="DNA ADP-ribosyl transferase"
FT /id="PRO_0000456050"
FT DOMAIN 29..230
FT /note="DarT"
FT /evidence="ECO:0000255"
FT REGION 54..70
FT /note="NAD(+)-binding element"
FT /evidence="ECO:0000250|UniProtKB:A0A0B0SG80"
FT REGION 135..183
FT /note="ADP-ribosylating turn-turn loop"
FT /evidence="ECO:0000250|UniProtKB:A0A0B0SG80"
FT ACT_SITE 67
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:A0A0B0SG80"
FT ACT_SITE 183
FT /evidence="ECO:0000250|UniProtKB:A0A0B0SG80"
FT BINDING 30..32
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:A0A0B0SG80"
FT BINDING 67
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:A0A0B0SG80"
SQ SEQUENCE 230 AA; 25580 MW; 1A109D34B16590A8 CRC64;
MITRYKPESG FVARSGGPDR KRPHDWIVWH FTHADNLPGI ITAGRLLADS AVTPTTEVAY
NPVKELRRHK VVAPDSRYPA SMASDHVPFY IAARSPMLYV VCKGHSGYSG GAGPLVHLGV
ALGDIIDADL TWCASDGNAA ASYTKFSRQV DTLGTFVDFD LLCQRQWHNT DDDPNRQSRR
AAEILVYGHV PFELVSYVCC YNTETMTRVR TLLDPVGGVR KYVIKPGMYY
