CAS9_CAMJE
ID CAS9_CAMJE Reviewed; 984 AA.
AC Q0P897;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 19-SEP-2006, sequence version 1.
DT 03-AUG-2022, entry version 70.
DE RecName: Full=CRISPR-associated endonuclease Cas9 {ECO:0000255|HAMAP-Rule:MF_01480};
GN Name=cas9 {ECO:0000255|HAMAP-Rule:MF_01480}; OrderedLocusNames=Cj1523c;
OS Campylobacter jejuni subsp. jejuni serotype O:2 (strain ATCC 700819 / NCTC
OS 11168).
OC Bacteria; Proteobacteria; Epsilonproteobacteria; Campylobacterales;
OC Campylobacteraceae; Campylobacter.
OX NCBI_TaxID=192222;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 700819 / NCTC 11168;
RX PubMed=10688204; DOI=10.1038/35001088;
RA Parkhill J., Wren B.W., Mungall K.L., Ketley J.M., Churcher C.M.,
RA Basham D., Chillingworth T., Davies R.M., Feltwell T., Holroyd S.,
RA Jagels K., Karlyshev A.V., Moule S., Pallen M.J., Penn C.W., Quail M.A.,
RA Rajandream M.A., Rutherford K.M., van Vliet A.H.M., Whitehead S.,
RA Barrell B.G.;
RT "The genome sequence of the food-borne pathogen Campylobacter jejuni
RT reveals hypervariable sequences.";
RL Nature 403:665-668(2000).
RN [2]
RP FUNCTION AS AN ENDONUCLEASE, AND POSSIBLE BIOTECHNOLOGY.
RC STRAIN=ATCC 700819 / NCTC 11168;
RX PubMed=24270795; DOI=10.1093/nar/gkt1074;
RA Fonfara I., Le Rhun A., Chylinski K., Makarova K.S., Lecrivain A.L.,
RA Bzdrenga J., Koonin E.V., Charpentier E.;
RT "Phylogeny of Cas9 determines functional exchangeability of dual-RNA and
RT Cas9 among orthologous type II CRISPR-Cas systems.";
RL Nucleic Acids Res. 42:2577-2590(2014).
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 spacers, sequences complementary to
CC antecedent mobile elements, and target invading nucleic acids. CRISPR
CC clusters are transcribed and processed into CRISPR RNA (crRNA). In type
CC II CRISPR systems correct processing of pre-crRNA requires a trans-
CC encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this
CC protein. The tracrRNA serves as a guide for ribonuclease 3-aided
CC processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA
CC endonucleolytically cleaves linear or circular dsDNA target
CC complementary to the spacer; Cas9 is inactive in the absence of the 2
CC guide RNAs (gRNA). Cas9 recognizes the protospacer adjacent motif (PAM)
CC in the CRISPR repeat sequences to help distinguish self versus nonself,
CC as targets within the bacterial CRISPR locus do not have PAMs. PAM
CC recognition is also required for catalytic activity (By similarity).
CC Cuts target DNA in Cas9:gRNAs mixing experiments with N.meningitidis
CC strain Z2491 and P.multocoda strain Pm70. {ECO:0000255|HAMAP-
CC Rule:MF_01480, ECO:0000269|PubMed:24270795}.
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_01480};
CC -!- SUBUNIT: Monomer. Binds crRNA and tracrRNA. {ECO:0000255|HAMAP-
CC Rule:MF_01480}.
CC -!- DOMAIN: Has 2 endonuclease domains. The discontinuous RuvC-like domain
CC cleaves the target DNA noncomplementary to crRNA while the HNH nuclease
CC domain cleaves the target DNA complementary to crRNA.
CC {ECO:0000255|HAMAP-Rule:MF_01480}.
CC -!- BIOTECHNOLOGY: The simplicity of the Cas9-gRNAs RNA-directed DNA
CC endonuclease activity may be used to target and modify a DNA sequence
CC of interest.
CC -!- SIMILARITY: Belongs to the CRISPR-associated protein Cas9 family.
CC Subtype II-C subfamily. {ECO:0000255|HAMAP-Rule:MF_01480}.
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DR EMBL; AL111168; CAL35627.1; -; Genomic_DNA.
DR PIR; F81299; F81299.
DR RefSeq; WP_002864485.1; NC_002163.1.
DR RefSeq; YP_002344900.1; NC_002163.1.
DR PDB; 5X2G; X-ray; 2.40 A; A=1-480, A=642-984.
DR PDB; 5X2H; X-ray; 2.30 A; A=1-480, A=642-984.
DR PDBsum; 5X2G; -.
DR PDBsum; 5X2H; -.
DR AlphaFoldDB; Q0P897; -.
DR SMR; Q0P897; -.
DR STRING; 192222.Cj1523c; -.
DR PaxDb; Q0P897; -.
DR PRIDE; Q0P897; -.
DR EnsemblBacteria; CAL35627; CAL35627; Cj1523c.
DR GeneID; 905809; -.
DR KEGG; cje:Cj1523c; -.
DR PATRIC; fig|192222.6.peg.1501; -.
DR eggNOG; COG3513; Bacteria.
DR HOGENOM; CLU_007514_0_0_7; -.
DR OMA; GQGHMET; -.
DR Proteomes; UP000000799; Chromosome.
DR GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR GO; GO:0004519; F:endonuclease activity; IEA:UniProtKB-UniRule.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0003723; F:RNA binding; IEA:UniProtKB-KW.
DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-UniRule.
DR GO; GO:0043571; P:maintenance of CRISPR repeat elements; IEA:UniProtKB-UniRule.
DR Gene3D; 3.30.420.10; -; 3.
DR HAMAP; MF_01480; Cas9; 1.
DR InterPro; IPR028629; Cas9.
DR InterPro; IPR033114; HNH_CAS9.
DR InterPro; IPR003615; HNH_nuc.
DR InterPro; IPR036397; RNaseH_sf.
DR InterPro; IPR041383; RuvC_III.
DR Pfam; PF13395; HNH_4; 1.
DR Pfam; PF18541; RuvC_III; 2.
DR TIGRFAMs; TIGR01865; cas_Csn1; 1.
DR PROSITE; PS51749; HNH_CAS9; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antiviral defense; DNA-binding; Endonuclease; Hydrolase;
KW Magnesium; Manganese; Metal-binding; Nuclease; Reference proteome;
KW RNA-binding.
FT CHAIN 1..984
FT /note="CRISPR-associated endonuclease Cas9"
FT /id="PRO_0000429983"
FT DOMAIN 487..637
FT /note="HNH Cas9-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01085"
FT ACT_SITE 8
FT /note="For RuvC-like nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT ACT_SITE 559
FT /note="Proton acceptor for HNH nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 8
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 8
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 479
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 483
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 483
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 707
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT STRAND 4..9
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 11..20
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 23..31
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 45..76
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 80..83
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 92..94
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 101..107
FT /evidence="ECO:0007829|PDB:5X2H"
FT TURN 108..110
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 115..127
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 144..156
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 159..161
FT /evidence="ECO:0007829|PDB:5X2G"
FT HELIX 162..169
FT /evidence="ECO:0007829|PDB:5X2H"
FT TURN 170..172
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 179..181
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 197..213
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 220..230
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 240..242
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 260..282
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 288..301
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 302..305
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 306..312
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 323..328
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 330..338
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 349..361
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 365..373
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 379..385
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 398..409
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 414..421
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 432..434
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 438..440
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 444..446
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 450..470
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 474..479
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 642..654
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 679..681
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 684..693
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 696..698
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 706..715
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 764..771
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 795..798
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 799..802
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 804..814
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 816..820
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 823..827
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 832..838
FT /evidence="ECO:0007829|PDB:5X2H"
FT TURN 839..841
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 844..849
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 851..856
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 882..890
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 893..897
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 905..913
FT /evidence="ECO:0007829|PDB:5X2H"
FT TURN 914..917
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 918..922
FT /evidence="ECO:0007829|PDB:5X2H"
FT HELIX 933..938
FT /evidence="ECO:0007829|PDB:5X2H"
FT TURN 939..941
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 948..953
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 958..965
FT /evidence="ECO:0007829|PDB:5X2H"
FT STRAND 971..973
FT /evidence="ECO:0007829|PDB:5X2H"
SQ SEQUENCE 984 AA; 114896 MW; 6194F6ACC05A1BFE CRC64;
MARILAFDIG ISSIGWAFSE NDELKDCGVR IFTKVENPKT GESLALPRRL ARSARKRLAR
RKARLNHLKH LIANEFKLNY EDYQSFDESL AKAYKGSLIS PYELRFRALN ELLSKQDFAR
VILHIAKRRG YDDIKNSDDK EKGAILKAIK QNEEKLANYQ SVGEYLYKEY FQKFKENSKE
FTNVRNKKES YERCIAQSFL KDELKLIFKK QREFGFSFSK KFEEEVLSVA FYKRALKDFS
HLVGNCSFFT DEKRAPKNSP LAFMFVALTR IINLLNNLKN TEGILYTKDD LNALLNEVLK
NGTLTYKQTK KLLGLSDDYE FKGEKGTYFI EFKKYKEFIK ALGEHNLSQD DLNEIAKDIT
LIKDEIKLKK ALAKYDLNQN QIDSLSKLEF KDHLNISFKA LKLVTPLMLE GKKYDEACNE
LNLKVAINED KKDFLPAFNE TYYKDEVTNP VVLRAIKEYR KVLNALLKKY GKVHKINIEL
AREVGKNHSQ RAKIEKEQNE NYKAKKDAEL ECEKLGLKIN SKNILKLRLF KEQKEFCAYS
GEKIKISDLQ DEKMLEIDHI YPYSRSFDDS YMNKVLVFTK QNQEKLNQTP FEAFGNDSAK
WQKIEVLAKN LPTKKQKRIL DKNYKDKEQK NFKDRNLNDT RYIARLVLNY TKDYLDFLPL
SDDENTKLND TQKGSKVHVE AKSGMLTSAL RHTWGFSAKD RNNHLHHAID AVIIAYANNS
IVKAFSDFKK EQESNSAELY AKKISELDYK NKRKFFEPFS GFRQKVLDKI DEIFVSKPER
KKPSGALHEE TFRKEEEFYQ SYGGKEGVLK ALELGKIRKV NGKIVKNGDM FRVDIFKHKK
TNKFYAVPIY TMDFALKVLP NKAVARSKKG EIKDWILMDE NYEFCFSLYK DSLILIQTKD
MQEPEFVYYN AFTSSTVSLI VSKHDNKFET LSKNQKILFK NANEKEVIAK SIGIQNLKVF
EKYIVSALGE VTKAEFRQRE DFKK
