THSA_BACCS
ID THSA_BACCS Reviewed; 476 AA.
AC J8G6Z1;
DT 03-AUG-2022, integrated into UniProtKB/Swiss-Prot.
DT 03-AUG-2022, sequence version 2.
DT 03-AUG-2022, entry version 31.
DE RecName: Full=NAD(+) hydrolase ThsA {ECO:0000303|PubMed:32499527};
DE Short=NADase ThsA {ECO:0000303|PubMed:34853457};
DE EC=3.2.2.5 {ECO:0000269|PubMed:32499527};
DE AltName: Full=Thoeris protein ThsA {ECO:0000303|PubMed:29371424};
GN Name=thsA {ECO:0000303|PubMed:29371424};
GN ORFNames=II9_05448 {ECO:0000312|EMBL:EJR09240.1};
OS Bacillus cereus (strain MSX-D12).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus;
OC Bacillus cereus group.
OX NCBI_TaxID=1053222;
RN [1] {ECO:0000312|EMBL:EJR09240.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=MSX-D12;
RG The Broad Institute Genome Sequencing Platform;
RG The Broad Institute Genome Sequencing Center for Infectious Disease;
RA Feldgarden M., Van der Auwera G.A., Mahillon J., Duprez V., Timmery S.,
RA Mattelet C., Dierick K., Sun M., Yu Z., Zhu L., Hu X., Shank E.B.,
RA Swiecicka I., Hansen B.M., Andrup L., Young S.K., Zeng Q., Gargeya S.,
RA Fitzgerald M., Haas B., Abouelleil A., Alvarado L., Arachchi H.M.,
RA Berlin A., Chapman S.B., Goldberg J., Griggs A., Gujja S., Hansen M.,
RA Howarth C., Imamovic A., Larimer J., McCowen C., Montmayeur A., Murphy C.,
RA Neiman D., Pearson M., Priest M., Roberts A., Saif S., Shea T., Sisk P.,
RA Sykes S., Wortman J., Nusbaum C., Birren B.;
RT "The Genome Sequence of Bacillus cereus MSX-D12.";
RL Submitted (APR-2012) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION IN ANTIVIRAL DEFENSE, SUBCELLULAR LOCATION, DISRUPTION PHENOTYPE,
RP EXPRESSION IN B.SUBTILIS, AND MUTAGENESIS OF ASN-112 AND ARG-371.
RC STRAIN=MSX-D12;
RX PubMed=29371424; DOI=10.1126/science.aar4120;
RA Doron S., Melamed S., Ofir G., Leavitt A., Lopatina A., Keren M.,
RA Amitai G., Sorek R.;
RT "Systematic discovery of antiphage defense systems in the microbial
RT pangenome.";
RL Science 359:0-0(2018).
RN [3]
RP FUNCTION IN ANTIVIRAL DEFENSE, FUNCTION AS AN NAD HYDROLASE, ACTIVITY
RP REGULATION, SUBUNIT, DOMAIN, AND MUTAGENESIS OF ASN-112.
RC STRAIN=MSX-D12;
RX PubMed=34853457; DOI=10.1038/s41586-021-04098-7;
RA Ofir G., Herbst E., Baroz M., Cohen D., Millman A., Doron S., Tal N.,
RA Malheiro D.B.A., Malitsky S., Amitai G., Sorek R.;
RT "Antiviral activity of bacterial TIR domains via immune signalling
RT molecules.";
RL Nature 600:116-120(2021).
RN [4] {ECO:0007744|PDB:6LHX}
RP X-RAY CRYSTALLOGRAPHY (2.50 ANGSTROMS), FUNCTION AS AN NAD HYDROLASE,
RP CATALYTIC ACTIVITY, PROBABLE ACTIVE SITE, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBUNIT, DOMAIN, AND MUTAGENESIS OF ASN-112 AND HIS-152.
RC STRAIN=MSX-D12;
RX PubMed=32499527; DOI=10.1038/s41467-020-16703-w;
RA Ka D., Oh H., Park E., Kim J.H., Bae E.;
RT "Structural and functional evidence of bacterial antiphage protection by
RT Thoeris defense system via NAD+ degradation.";
RL Nat. Commun. 11:2816-2816(2020).
CC -!- FUNCTION: NAD(+) hydrolyzing component of antiviral defense system
CC Thoeris, composed of ThsA and ThsB. Activated by a signal molecule
CC generated by ThsB, by TIR1 and TIR2 from B.dafuensis or by BdTIR from
CC B.distachyon. Upon activation binds and hydrolyzes NAD(+), leading to
CC cell death and inhibition of phage replication. Not seen to bind DNA
CC (PubMed:32499527, PubMed:34853457). Expression of ThsA and ThsB in
CC B.subtilis (strain BEST7003) confers resistance to phages phi29,
CC SBSphiC, SBSphiJ and SPO1 (PubMed:29371424, PubMed:34853457). At
CC multiplicity of infection (MOI) of 0.05 Thoeris-encoding cultures grow
CC normally when infected with SPO1, at MOI 5 cultures collapse
CC prematurely by 90 minutes post-infection, thus the phage are not able
CC to complete a replication cycle. NAD(+) levels fall and ADP-D-ribose
CC levels rise 60 minutes post-infection. Thoeris cultures eventually
CC recover, but retain the same susceptibility to SPO1 (PubMed:34853457).
CC {ECO:0000269|PubMed:29371424, ECO:0000269|PubMed:32499527,
CC ECO:0000269|PubMed:34853457}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + NAD(+) = ADP-D-ribose + H(+) + nicotinamide;
CC Xref=Rhea:RHEA:16301, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:17154, ChEBI:CHEBI:57540, ChEBI:CHEBI:57967; EC=3.2.2.5;
CC Evidence={ECO:0000269|PubMed:32499527};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:16302;
CC Evidence={ECO:0000269|PubMed:32499527};
CC -!- ACTIVITY REGULATION: Activated by a signal molecule generated by
CC endogenous ThsB, by TIR1 (AC A0A5B8Z670) and TIR2 (AC A0A5B8Z260) of
CC B.dafuensis, and by BdTIR (AC I1GTC2), a plant protein involved in
CC defense (PubMed:34853457). Activation may alter the oligomerization
CC state of the protein (PubMed:34853457) (Probable).
CC {ECO:0000269|PubMed:34853457, ECO:0000305|PubMed:34853457}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=269.8 uM for NAD(+) {ECO:0000269|PubMed:32499527};
CC Note=kcat is 33.9 min(-1) (PubMed:32499527).
CC {ECO:0000269|PubMed:32499527};
CC -!- SUBUNIT: Homotetramer formed by dimer of dimers; homooctamers are
CC occasionally seen. Not seen to interact with ThsB (PubMed:32499527). In
CC the absence of the signal generated by ThsB, 63% monomer and 20%
CC homotetramer; in the presence of the ThsB signal product 40% of the
CC protein is dimeric (PubMed:34853457). {ECO:0000269|PubMed:32499527,
CC ECO:0000269|PubMed:34853457}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305|PubMed:29371424}.
CC -!- DOMAIN: Has an N-terminal sirtuin-like domain (SIR2, residues 1-283)
CC and a C-terminal SLOG (STALD)-like domain (residues 284-476)
CC (PubMed:32499527). The SIR2 domain has NAD(+) hydrolase activity
CC (PubMed:32499527, PubMed:34853457). The C-terminus probably drives
CC oligomerization which activates the NADase activity of the N-terminus
CC (PubMed:34853457) (Probable). {ECO:0000269|PubMed:32499527,
CC ECO:0000269|PubMed:34853457, ECO:0000305|PubMed:34853457}.
CC -!- DISRUPTION PHENOTYPE: When this gene is missing the Thoeris system does
CC not confer phage resistance in B.subtilis.
CC {ECO:0000269|PubMed:29371424}.
CC -!- SIMILARITY: Belongs to the Thoeris B TIR-like family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=EJR09240.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305|PubMed:29371424};
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; AHEQ01000050; EJR09240.1; ALT_INIT; Genomic_DNA.
DR PDB; 6LHX; X-ray; 2.50 A; A/B/C/D=3-476.
DR PDBsum; 6LHX; -.
DR SMR; J8G6Z1; -.
DR EnsemblBacteria; EJR09240; EJR09240; II9_05448.
DR PATRIC; fig|1053222.3.peg.5515; -.
DR HOGENOM; CLU_028011_0_0_9; -.
DR Proteomes; UP000006971; Unassembled WGS sequence.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR InterPro; IPR029035; DHS-like_NAD/FAD-binding_dom.
DR InterPro; IPR039444; SIR2-like.
DR InterPro; IPR041486; STALD.
DR Pfam; PF13289; SIR2_2; 1.
DR Pfam; PF18185; STALD; 1.
DR SUPFAM; SSF52467; SSF52467; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antiviral defense; Cytoplasm; Hydrolase.
FT CHAIN 1..476
FT /note="NAD(+) hydrolase ThsA"
FT /id="PRO_0000456257"
FT ACT_SITE 152
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:32499527"
FT SITE 112
FT /note="NAD(+) recognition"
FT /evidence="ECO:0000305|PubMed:32499527"
FT MUTAGEN 112
FT /note="N->A: No longer confers resistance to phage. Loss of
FT NAD(+) hydrolase activity, no longer binds NAD(+),
FT oligomerizes correctly. No change in NAD(+) levels during
FT SPO1 infection, still dimerizes in response to signal."
FT /evidence="ECO:0000269|PubMed:29371424,
FT ECO:0000269|PubMed:32499527, ECO:0000269|PubMed:34853457"
FT MUTAGEN 152
FT /note="H->A: Loss of NAD(+) hydrolase activity, does not
FT oligomerize correctly."
FT /evidence="ECO:0000269|PubMed:32499527"
FT MUTAGEN 371
FT /note="R->A: No resistance to phage SPO1, no
FT oligomerization in absence of signal, a little bit of dimer
FT seen in response to signal."
FT /evidence="ECO:0000269|PubMed:29371424"
SQ SEQUENCE 476 AA; 55190 MW; 9EC43136AE8FDACB CRC64;
MKMNPIVELF IKDFTKEVME ENAAIFAGAG LSMSVGYVSW AKLLEPIAQE IGLDVNKEND
LVSLAQYYCN ENQGNRGRIN QIILDEFSRK VDLTENHKIL ARLPIHTYWT TNYDRLIEKA
LEEENKIADV KYTVKQLATT KVKRDAVVYK MHGDVEHPSE AVLIKDDYEK YSIKMDPYIK
ALSGDLVSKT FLFVGFSFTD PNLDYILSRV RSAYERDQRR HYCLIKKEER RPDELEADFE
YRVRKQELFI SDLSRFNIKT IVLNNYNEIT EILQRIENNI KTKTVFLSGS AVEYNHWETE
HAEQFIHQLS KELIRKDFNI VSGFGLGVGS FVINGVLEEL YMNQGTIDDD RLILRPFPQG
KKGEEQWDKY RRDMITRTGV SIFLYGNKID KGQVVKAKGV QSEFNISFEQ NNYVVPVGAT
GYIAKDLWNK VNEEFETYYP GADARMKKLF GELNNEALSI EELINTIIEF VEILSN
