CS3HD_METJA
ID CS3HD_METJA Reviewed; 244 AA.
AC Q57829;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 93.
DE RecName: Full=CRISPR-associated endonuclease Cas3-HD;
DE EC=3.1.-.-;
DE AltName: Full=CRISPR-associated ssDNA/ssRNA endonuclease Cas3-HD;
GN Name=cas3; Synonyms=cas3''; OrderedLocusNames=MJ0384;
OS Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM
OS 10045 / NBRC 100440) (Methanococcus jannaschii).
OC Archaea; Euryarchaeota; Methanomada group; Methanococci; Methanococcales;
OC Methanocaldococcaceae; Methanocaldococcus.
OX NCBI_TaxID=243232;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440;
RX PubMed=8688087; DOI=10.1126/science.273.5278.1058;
RA Bult C.J., White O., Olsen G.J., Zhou L., Fleischmann R.D., Sutton G.G.,
RA Blake J.A., FitzGerald L.M., Clayton R.A., Gocayne J.D., Kerlavage A.R.,
RA Dougherty B.A., Tomb J.-F., Adams M.D., Reich C.I., Overbeek R.,
RA Kirkness E.F., Weinstock K.G., Merrick J.M., Glodek A., Scott J.L.,
RA Geoghagen N.S.M., Weidman J.F., Fuhrmann J.L., Nguyen D., Utterback T.R.,
RA Kelley J.M., Peterson J.D., Sadow P.W., Hanna M.C., Cotton M.D.,
RA Roberts K.M., Hurst M.A., Kaine B.P., Borodovsky M., Klenk H.-P.,
RA Fraser C.M., Smith H.O., Woese C.R., Venter J.C.;
RT "Complete genome sequence of the methanogenic archaeon, Methanococcus
RT jannaschii.";
RL Science 273:1058-1073(1996).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS), FUNCTION AS A NUCLEASE, COFACTOR,
RP SUBSTRATES, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, SUBUNIT,
RP AND MUTAGENESIS OF HIS-20; HIS-66; ASP-67; HIS-91; HIS-123; HIS-124 AND
RP ASP-219.
RC STRAIN=ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440;
RX PubMed=22009198; DOI=10.1038/emboj.2011.377;
RA Beloglazova N., Petit P., Flick R., Brown G., Savchenko A., Yakunin A.F.;
RT "Structure and activity of the Cas3 HD nuclease MJ0384, an effector enzyme
RT of the CRISPR interference.";
RL EMBO J. 30:4616-4627(2011).
CC -!- FUNCTION: CRISPR (clustered regularly interspaced short palindromic
CC repeat), is an adaptive immune system that provides protection against
CC mobile genetic elements (viruses, transposable elements and conjugative
CC plasmids). CRISPR clusters contain sequences complementary to
CC antecedent mobile elements and target invading nucleic acids. CRISPR
CC clusters are transcribed and processed into CRISPR RNA (crRNA). Cas3
CC plus Cascade participate in CRISPR interference, the third stage of
CC CRISPR immunity. Acts as a ssDNA and ssRNA nuclease, with both
CC endo- and 3' to 5' exonuclease activities, acting on substrates with
CC free single-stranded 3' ends. Double-stranded nucleic acids are not
CC substrates. Activity is higher for DNA than RNA. Templates include R-
CC loops (a bubble-like structure formed when ssRNA replaces one strand in
CC a dsDNA, such as crRNA is thought to form with CRISPR target DNA),
CC circular ssDNA, 2',3'-cAMP and 2',3'-cGMP. Probably generates 3'-
CC phosphate and 5'-hydroxyl ends. In the presence of the Cas3 helicase
CC and ATP dsDNA templates are also degraded.
CC {ECO:0000269|PubMed:22009198}.
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:22009198};
CC Note=ssDNase activity requires Mg(2+), ssRNase activity does not
CC require cations. A second loosely associated metal ion is visible in
CC the crystal structure. {ECO:0000269|PubMed:22009198};
CC -!- ACTIVITY REGULATION: Both ssDNase and ssRNase are inhibited by EDTA.
CC {ECO:0000269|PubMed:22009198}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 7-9, varying somewhat depending on the substrate.
CC {ECO:0000269|PubMed:22009198};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:22009198}.
CC -!- DOMAIN: Proteins of this family have an N-terminal nuclease domain and
CC a C-terminal helicase/ATPase domain. In some CRISPR/Cas systems the
CC domains are swapped, in others they are encoded separately.
CC -!- SIMILARITY: Belongs to the CRISPR-associated nuclease Cas3-HD family.
CC {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; L77117; AAB98379.1; -; Genomic_DNA.
DR PIR; H64347; H64347.
DR PDB; 3S4L; X-ray; 2.30 A; A=1-244.
DR PDBsum; 3S4L; -.
DR AlphaFoldDB; Q57829; -.
DR SMR; Q57829; -.
DR STRING; 243232.MJ_0384; -.
DR DNASU; 1451241; -.
DR EnsemblBacteria; AAB98379; AAB98379; MJ_0384.
DR KEGG; mja:MJ_0384; -.
DR eggNOG; arCOG01442; Archaea.
DR HOGENOM; CLU_1052127_0_0_2; -.
DR OMA; HEPILMG; -.
DR PhylomeDB; Q57829; -.
DR EvolutionaryTrace; Q57829; -.
DR Proteomes; UP000000805; Chromosome.
DR GO; GO:0004519; F:endonuclease activity; IEA:UniProtKB-KW.
DR GO; GO:0004527; F:exonuclease activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-KW.
DR Gene3D; 1.10.3210.30; -; 1.
DR InterPro; IPR006483; CRISPR-assoc_Cas3_HD.
DR InterPro; IPR038257; CRISPR-assoc_Cas3_HD_sf.
DR Pfam; PF18019; HD_6; 1.
DR TIGRFAMs; TIGR01596; cas3_HD; 1.
DR PROSITE; PS51643; HD_CAS3; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antiviral defense; Endonuclease; Exonuclease; Hydrolase;
KW Magnesium; Metal-binding; Nuclease; Reference proteome.
FT CHAIN 1..244
FT /note="CRISPR-associated endonuclease Cas3-HD"
FT /id="PRO_0000106847"
FT DOMAIN 10..221
FT /note="HD Cas3-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00974"
FT BINDING 67
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT BINDING 91
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT BINDING 123
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT BINDING 124
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT MUTAGEN 20
FT /note="H->A: Loss of all nuclease activity."
FT /evidence="ECO:0000269|PubMed:22009198"
FT MUTAGEN 66
FT /note="H->A: Loss of all nuclease activity."
FT /evidence="ECO:0000269|PubMed:22009198"
FT MUTAGEN 67
FT /note="D->A: Loss of all nuclease activity."
FT /evidence="ECO:0000269|PubMed:22009198"
FT MUTAGEN 67
FT /note="D->E: Very little nuclease activity."
FT /evidence="ECO:0000269|PubMed:22009198"
FT MUTAGEN 91
FT /note="H->A: Very little ssDNA exonuclease activity."
FT /evidence="ECO:0000269|PubMed:22009198"
FT MUTAGEN 123
FT /note="H->A: Very little nuclease activity."
FT /evidence="ECO:0000269|PubMed:22009198"
FT MUTAGEN 124
FT /note="H->A: Very little nuclease activity."
FT /evidence="ECO:0000269|PubMed:22009198"
FT MUTAGEN 219
FT /note="D->A: Loss of all nuclease activity."
FT /evidence="ECO:0000269|PubMed:22009198"
FT STRAND 10..12
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 17..31
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 32..34
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 36..43
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 44..46
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 52..65
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 68..71
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 73..80
FT /evidence="ECO:0007829|PDB:3S4L"
FT STRAND 82..84
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 91..107
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 110..121
FT /evidence="ECO:0007829|PDB:3S4L"
FT TURN 122..124
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 142..149
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 159..167
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 181..197
FT /evidence="ECO:0007829|PDB:3S4L"
FT TURN 201..203
FT /evidence="ECO:0007829|PDB:3S4L"
FT HELIX 204..215
FT /evidence="ECO:0007829|PDB:3S4L"
SQ SEQUENCE 244 AA; 28406 MW; 09CE6E74BB8621C3 CRC64;
MILGEIMEVL AFKNQSLIDH VNDMVKYWER IKYRYLKTIK RALEALNIKL DIEKVDEFMK
ILIKLHDIGK ASKIYQRAII NDQEKLMGFR HELVSAYYTY HILLKKFGDK NLAFIGALTV
MLHHEPIIMG QIRNLKKKEL TAEVVLDKLK KFDGMIEDFE DLIKKLIGYS IGDIIKNDSN
KDDIIRFVIE MSVRARHTPN SEKLRFIVGT LLLPLVMCDY KGAESREGKA PKFAEVLEVE
SYVI
