MBCT_MYCTU
ID MBCT_MYCTU Reviewed; 186 AA.
AC P9WLP9; L0TB64; P64907; Q10869;
DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot.
DT 16-APR-2014, sequence version 1.
DT 03-AUG-2022, entry version 33.
DE RecName: Full=NAD(+) phosphorylase MbcT {ECO:0000303|PubMed:30792174};
DE EC=2.4.2.- {ECO:0000269|PubMed:30792174};
DE AltName: Full=Mycobacterial cidal toxin MbcT {ECO:0000303|PubMed:30792174};
DE AltName: Full=Toxin Res {ECO:0000303|PubMed:30315706};
DE AltName: Full=ucAT8 toxin {ECO:0000303|PubMed:28724903};
GN Name=mbcT {ECO:0000303|PubMed:30792174}; OrderedLocusNames=Rv1989c;
GN ORFNames=MTCY39.30;
OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae;
OC Mycobacterium; Mycobacterium tuberculosis complex.
OX NCBI_TaxID=83332;
RN [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]
RP INDUCTION BY HYPOXIA.
RC STRAIN=ATCC 27294 / TMC 102 / H37Rv;
RX PubMed=18231589; DOI=10.1371/journal.pone.0001502;
RA Rustad T.R., Harrell M.I., Liao R., Sherman D.R.;
RT "The enduring hypoxic response of Mycobacterium tuberculosis.";
RL PLoS ONE 3:E1502-E1502(2008).
RN [3]
RP INDUCTION IN MOUSE INFECTION MODEL.
RC STRAIN=H37Rv;
RX PubMed=20628579; DOI=10.1371/journal.ppat.1000988;
RA Homolka S., Niemann S., Russell D.G., Rohde K.H.;
RT "Functional genetic diversity among Mycobacterium tuberculosis complex
RT clinical isolates: delineation of conserved core and lineage-specific
RT transcriptomes during intracellular survival.";
RL PLoS Pathog. 6:E1000988-E1000988(2010).
RN [4]
RP INDUCTION BY HYPOXIA AND IN PERSISTER CELLS.
RC STRAIN=H37Rv;
RX PubMed=21673191; DOI=10.1128/mbio.00100-11;
RA Keren I., Minami S., Rubin E., Lewis K.;
RT "Characterization and transcriptome analysis of Mycobacterium tuberculosis
RT persisters.";
RL MBio 2:E00100-E00111(2011).
RN [5]
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 [6]
RP DISRUPTION PHENOTYPE.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=28096490; DOI=10.1128/mbio.02133-16;
RA DeJesus M.A., Gerrick E.R., Xu W., Park S.W., Long J.E., Boutte C.C.,
RA Rubin E.J., Schnappinger D., Ehrt S., Fortune S.M., Sassetti C.M.,
RA Ioerger T.R.;
RT "Comprehensive essentiality analysis of the Mycobacterium tuberculosis
RT genome via saturating transposon mutagenesis.";
RL MBio 8:0-0(2017).
RN [7]
RP INDUCTION BY STRESS.
RC STRAIN=H37Rv;
RX PubMed=28724903; DOI=10.1038/s41598-017-06003-7;
RA Gupta A., Venkataraman B., Vasudevan M., Gopinath Bankar K.;
RT "Co-expression network analysis of toxin-antitoxin loci in Mycobacterium
RT tuberculosis reveals key modulators of cellular stress.";
RL Sci. Rep. 7:5868-5868(2017).
RN [8]
RP FUNCTION AS A TOXIN, AND EXPRESSION IN E.COLI.
RC STRAIN=H37Rv;
RX PubMed=30315706; DOI=10.1111/mmi.14150;
RA Skjerning R.B., Senissar M., Winther K.S., Gerdes K., Brodersen D.E.;
RT "The RES domain toxins of RES-Xre toxin-antitoxin modules induce cell
RT stasis by degrading NAD+.";
RL Mol. Microbiol. 111:221-236(2019).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF 2-186 IN COMPLEX WITH ANTITOXIN,
RP FUNCTION AS A TOXIN, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBUNIT, INDUCTION BY STARVATION, DISRUPTION PHENOTYPE, BIOTECHNOLOGY, AND
RP MUTAGENESIS OF ARG-27; TYR-28; ARG-47; TYR-58; GLU-69 AND SER-126.
RC STRAIN=ATCC 27294 / TMC 102 / H37Rv;
RX PubMed=30792174; DOI=10.1016/j.molcel.2019.01.028;
RA Freire D.M., Gutierrez C., Garza-Garcia A., Grabowska A.D., Sala A.J.,
RA Ariyachaokun K., Panikova T., Beckham K.S.H., Colom A., Pogenberg V.,
RA Cianci M., Tuukkanen A., Boudehen Y.M., Peixoto A., Botella L.,
RA Svergun D.I., Schnappinger D., Schneider T.R., Genevaux P.,
RA de Carvalho L.P.S., Wilmanns M., Parret A.H.A., Neyrolles O.;
RT "An NAD+ phosphorylase toxin triggers Mycobacterium tuberculosis cell
RT death.";
RL Mol. Cell 73:1282-1291(2019).
CC -!- FUNCTION: Toxic component of a type II toxin-antitoxin (TA) system.
CC Neutralized by cognate antitoxin MbcA (PubMed:30315706,
CC PubMed:30792174). Degrades NAD(+) by phosphorolysis. Expression in the
CC absence of its cognate antitoxin MbcA causes dramatic reduction of
CC intracellular NAD(+) levels and is deleterious to cell growth, causing
CC cell death. In a SCID mouse infection model, mice infected with
CC bacteria overexpressing this protein survive longer. Overexpression of
CC this protein in a mouse infection model at 21 days leads to bacterial
CC death, and shows a synergistic 100-fold increase in mouse survival when
CC combined with isoniazid treatment (PubMed:30792174).
CC {ECO:0000269|PubMed:30315706, ECO:0000269|PubMed:30792174}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=NAD(+) + phosphate = ADP-beta-D-ribose 1''-phosphate + H(+) +
CC nicotinamide; Xref=Rhea:RHEA:20788, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:17154, ChEBI:CHEBI:43474, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:58753; Evidence={ECO:0000269|PubMed:30792174};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=110 uM for NAD(+) {ECO:0000269|PubMed:30792174};
CC Note=kcat for NAD(+) is 167 sec(-1). {ECO:0000269|PubMed:30792174};
CC -!- SUBUNIT: Heterotetramer with 2 subunits each of MbcT and MbcA; the
CC tetramers further assemble into trimers with 3-fold symmetry. The
CC antitoxin acts by blocking acces to the toxin active site.
CC {ECO:0000269|PubMed:30792174}.
CC -!- INDUCTION: Induced by hypoxia (PubMed:18231589). Expression induced in
CC both active and resting C57BL/6 mouse macrophages (PubMed:20628579).
CC Induced in persister cells (PubMed:21673191). Induced by streptomycin
CC treatment and by starvation (PubMed:28724903). Induced by starvation
CC stress; probably part of the mbcT-mbcA operon (PubMed:30792174).
CC {ECO:0000269|PubMed:18231589, ECO:0000269|PubMed:20628579,
CC ECO:0000269|PubMed:21673191, ECO:0000269|PubMed:28724903,
CC ECO:0000269|PubMed:30792174}.
CC -!- DISRUPTION PHENOTYPE: Not essential, it can be deleted
CC (PubMed:28096490). Deletion of the mbcT-mbcA operon has no visible
CC phenotype (PubMed:30792174). {ECO:0000269|PubMed:28096490,
CC ECO:0000269|PubMed:30792174}.
CC -!- BIOTECHNOLOGY: Molecules that disrupt the MbcT-MbcA complex could be
CC candidates for anti-tuberculosis therapy.
CC {ECO:0000269|PubMed:30792174}.
CC -!- SIMILARITY: Belongs to the MbcT/ParT/Res family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; AL123456; CCP44759.1; -; Genomic_DNA.
DR PIR; C70757; C70757.
DR RefSeq; NP_216505.1; NC_000962.3.
DR RefSeq; WP_003410001.1; NZ_NVQJ01000043.1.
DR PDB; 6FKG; X-ray; 1.80 A; A/B=2-186.
DR PDBsum; 6FKG; -.
DR AlphaFoldDB; P9WLP9; -.
DR SASBDB; P9WLP9; -.
DR SMR; P9WLP9; -.
DR STRING; 83332.Rv1989c; -.
DR PaxDb; P9WLP9; -.
DR DNASU; 885810; -.
DR GeneID; 885810; -.
DR KEGG; mtu:Rv1989c; -.
DR TubercuList; Rv1989c; -.
DR eggNOG; COG5654; Bacteria.
DR OMA; DSTRACM; -.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0016779; F:nucleotidyltransferase activity; IEA:UniProtKB-KW.
DR InterPro; IPR014914; RES_dom.
DR Pfam; PF08808; RES; 1.
DR SMART; SM00953; RES; 1.
PE 1: Evidence at protein level;
KW 3D-structure; NAD; Nucleotidyltransferase; Reference proteome;
KW Toxin-antitoxin system; Transferase.
FT CHAIN 1..186
FT /note="NAD(+) phosphorylase MbcT"
FT /id="PRO_0000103918"
FT MUTAGEN 27
FT /note="R->A: No longer toxic."
FT /evidence="ECO:0000269|PubMed:30792174"
FT MUTAGEN 27
FT /note="R->E: No longer toxic, does not degrade NAD(+)."
FT /evidence="ECO:0000269|PubMed:30792174"
FT MUTAGEN 28
FT /note="Y->A: Reduced toxicity."
FT /evidence="ECO:0000269|PubMed:30792174"
FT MUTAGEN 47
FT /note="R->A: No longer toxic."
FT /evidence="ECO:0000269|PubMed:30792174"
FT MUTAGEN 58
FT /note="Y->A: No longer toxic."
FT /evidence="ECO:0000269|PubMed:30792174"
FT MUTAGEN 69
FT /note="E->A: Reduced toxicity."
FT /evidence="ECO:0000269|PubMed:30792174"
FT MUTAGEN 126
FT /note="S->A: No change in toxicity."
FT /evidence="ECO:0000269|PubMed:30792174"
FT HELIX 7..16
FT /evidence="ECO:0007829|PDB:6FKG"
FT STRAND 19..30
FT /evidence="ECO:0007829|PDB:6FKG"
FT HELIX 39..44
FT /evidence="ECO:0007829|PDB:6FKG"
FT STRAND 56..62
FT /evidence="ECO:0007829|PDB:6FKG"
FT HELIX 63..76
FT /evidence="ECO:0007829|PDB:6FKG"
FT HELIX 81..84
FT /evidence="ECO:0007829|PDB:6FKG"
FT STRAND 89..103
FT /evidence="ECO:0007829|PDB:6FKG"
FT HELIX 107..112
FT /evidence="ECO:0007829|PDB:6FKG"
FT HELIX 117..121
FT /evidence="ECO:0007829|PDB:6FKG"
FT HELIX 126..137
FT /evidence="ECO:0007829|PDB:6FKG"
FT STRAND 142..147
FT /evidence="ECO:0007829|PDB:6FKG"
FT STRAND 150..157
FT /evidence="ECO:0007829|PDB:6FKG"
FT HELIX 158..161
FT /evidence="ECO:0007829|PDB:6FKG"
FT STRAND 166..174
FT /evidence="ECO:0007829|PDB:6FKG"
FT HELIX 177..184
FT /evidence="ECO:0007829|PDB:6FKG"
SQ SEQUENCE 186 AA; 20281 MW; 8414F65A43BD909D CRC64;
MSDALDEGLV QRIDARGTIE WSETCYRYTG AHRDALSGEG ARRFGGRWNP PLLFPAIYLA
DSAQACMVEV ERAAQAASTT AEKMLEAAYR LHTIDVTDLA VLDLTTPQAR EAVGLENDDI
YGDDWSGCQA VGHAAWFLHM QGVLVPAAGG VGLVVTAYEQ RTRPGQLQLR QSVDLTPALY
QELRAT
