CAS9_CORDI
ID CAS9_CORDI Reviewed; 1084 AA.
AC Q6NKI3;
DT 11-MAY-2016, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 87.
DE RecName: Full=CRISPR-associated endonuclease Cas9 {ECO:0000303|PubMed:25830891};
DE EC=3.1.-.-;
DE AltName: Full=SaCas9 {ECO:0000303|PubMed:25830891};
GN Name=cas9 {ECO:0000303|PubMed:25830891}; OrderedLocusNames=DIP0036;
OS Corynebacterium diphtheriae (strain ATCC 700971 / NCTC 13129 / Biotype
OS gravis).
OC Bacteria; Actinobacteria; Corynebacteriales; Corynebacteriaceae;
OC Corynebacterium.
OX NCBI_TaxID=257309;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 700971 / NCTC 13129 / Biotype gravis;
RX PubMed=14602910; DOI=10.1093/nar/gkg874;
RA Cerdeno-Tarraga A.-M., Efstratiou A., Dover L.G., Holden M.T.G.,
RA Pallen M.J., Bentley S.D., Besra G.S., Churcher C.M., James K.D.,
RA De Zoysa A., Chillingworth T., Cronin A., Dowd L., Feltwell T., Hamlin N.,
RA Holroyd S., Jagels K., Moule S., Quail M.A., Rabbinowitsch E.,
RA Rutherford K.M., Thomson N.R., Unwin L., Whitehead S., Barrell B.G.,
RA Parkhill J.;
RT "The complete genome sequence and analysis of Corynebacterium diphtheriae
RT NCTC13129.";
RL Nucleic Acids Res. 31:6516-6523(2003).
RN [2]
RP FUNCTION, AND POSSIBLE BIOTECHNOLOGY.
RX PubMed=25830891; DOI=10.1038/nature14299;
RA Ran F.A., Cong L., Yan W.X., Scott D.A., Gootenberg J.S., Kriz A.J.,
RA Zetsche B., Shalem O., Wu X., Makarova K.S., Koonin E.V., Sharp P.A.,
RA Zhang F.;
RT "In vivo genome editing using Staphylococcus aureus Cas9.";
RL Nature 520:186-191(2015).
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) (PubMed:25830891). Cas9 recognizes the protospacer
CC adjacent motif (PAM) in the CRISPR repeat sequences to help distinguish
CC self versus nonself, as targets within the bacterial CRISPR locus do
CC not have PAMs (PubMed:25830891). PAM recognition is also required for
CC catalytic activity (By similarity). {ECO:0000250|UniProtKB:Q99ZW2,
CC ECO:0000269|PubMed:25830891}.
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000250|UniProtKB:Q99ZW2};
CC Note=Endonuclease activity on target dsDNA requires Mg(2+).
CC {ECO:0000250|UniProtKB:Q99ZW2};
CC -!- SUBUNIT: Monomer. Binds crRNA and tracrRNA (Probable).
CC {ECO:0000305|PubMed:25830891}.
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:0000250|UniProtKB:Q99ZW2}.
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; this enzyme has been tested in human cells but is not
CC highly efficient. {ECO:0000269|PubMed:25830891}.
CC -!- SIMILARITY: Belongs to the CRISPR-associated protein Cas9 family.
CC Subtype II-C subfamily. {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; BX248354; CAE48549.1; -; Genomic_DNA.
DR RefSeq; WP_010933968.1; NC_002935.2.
DR PDB; 6JOO; X-ray; 2.90 A; A=1-497, A=664-1084.
DR PDBsum; 6JOO; -.
DR AlphaFoldDB; Q6NKI3; -.
DR SMR; Q6NKI3; -.
DR STRING; 257309.DIP0036; -.
DR PRIDE; Q6NKI3; -.
DR EnsemblBacteria; CAE48549; CAE48549; DIP0036.
DR KEGG; cdi:DIP0036; -.
DR HOGENOM; CLU_280423_0_0_11; -.
DR OMA; ESATKTW; -.
DR OrthoDB; 24005at2; -.
DR Proteomes; UP000002198; Chromosome.
DR GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR GO; GO:0004519; F:endonuclease activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0003723; F:RNA binding; IEA:UniProtKB-KW.
DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-KW.
DR Gene3D; 3.30.420.10; -; 3.
DR InterPro; IPR040619; Cas9_alpha-helical_lobe.
DR InterPro; IPR040796; Cas9_b_hairpin.
DR InterPro; IPR041217; Cas9_C.
DR InterPro; IPR041225; Cas9_Topo.
DR InterPro; IPR002711; HNH.
DR InterPro; IPR033114; HNH_CAS9.
DR InterPro; IPR003615; HNH_nuc.
DR InterPro; IPR036397; RNaseH_sf.
DR InterPro; IPR041383; RuvC_III.
DR Pfam; PF18470; Cas9_a; 1.
DR Pfam; PF17893; Cas9_b_hairpin; 1.
DR Pfam; PF18525; Cas9_C; 1.
DR Pfam; PF17894; Cas9_Topo; 1.
DR Pfam; PF01844; HNH; 1.
DR Pfam; PF18541; RuvC_III; 1.
DR SMART; SM00507; HNHc; 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..1084
FT /note="CRISPR-associated endonuclease Cas9"
FT /id="PRO_0000436102"
FT DOMAIN 504..665
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:0000250|UniProtKB:Q99ZW2"
FT ACT_SITE 573
FT /note="Proton acceptor for HNH nuclease domain"
FT /evidence="ECO:0000250|UniProtKB:Q99ZW2"
FT BINDING 8
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q99ZW2"
FT BINDING 8
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q99ZW2"
FT BINDING 496
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q99ZW2"
FT BINDING 500
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q99ZW2"
FT BINDING 500
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q99ZW2"
FT BINDING 727
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q99ZW2"
FT STRAND 3..9
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 11..21
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 23..25
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 27..37
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 53..83
FT /evidence="ECO:0007829|PDB:6JOO"
FT TURN 91..93
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 99..107
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 115..131
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 142..145
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 153..166
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 176..181
FT /evidence="ECO:0007829|PDB:6JOO"
FT TURN 193..195
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 197..199
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 203..216
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 221..231
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 238..241
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 249..251
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 258..261
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 262..274
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 293..302
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 345..352
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 356..364
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 367..377
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 385..387
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 388..399
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 402..410
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 422..433
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 441..444
FT /evidence="ECO:0007829|PDB:6JOO"
FT TURN 445..448
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 467..487
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 491..496
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 669..682
FT /evidence="ECO:0007829|PDB:6JOO"
FT TURN 683..687
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 689..693
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 695..699
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 714..718
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 721..723
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 726..734
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 739..748
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 772..797
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 802..804
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 825..828
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 835..838
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 841..843
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 844..851
FT /evidence="ECO:0007829|PDB:6JOO"
FT TURN 858..860
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 870..872
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 875..877
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 881..885
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 887..890
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 892..895
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 898..901
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 906..914
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 921..929
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 930..932
FT /evidence="ECO:0007829|PDB:6JOO"
FT TURN 934..937
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 940..942
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 950..953
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 957..965
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 968..974
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 979..981
FT /evidence="ECO:0007829|PDB:6JOO"
FT TURN 984..987
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 990..999
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 1004..1012
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 1015..1020
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 1025..1027
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 1034..1040
FT /evidence="ECO:0007829|PDB:6JOO"
FT HELIX 1049..1055
FT /evidence="ECO:0007829|PDB:6JOO"
FT STRAND 1059..1061
FT /evidence="ECO:0007829|PDB:6JOO"
SQ SEQUENCE 1084 AA; 121522 MW; D565E762D21C1EE3 CRC64;
MKYHVGIDVG TFSVGLAAIE VDDAGMPIKT LSLVSHIHDS GLDPDEIKSA VTRLASSGIA
RRTRRLYRRK RRRLQQLDKF IQRQGWPVIE LEDYSDPLYP WKVRAELAAS YIADEKERGE
KLSVALRHIA RHRGWRNPYA KVSSLYLPDG PSDAFKAIRE EIKRASGQPV PETATVGQMV
TLCELGTLKL RGEGGVLSAR LQQSDYAREI QEICRMQEIG QELYRKIIDV VFAAESPKGS
ASSRVGKDPL QPGKNRALKA SDAFQRYRIA ALIGNLRVRV DGEKRILSVE EKNLVFDHLV
NLTPKKEPEW VTIAEILGID RGQLIGTATM TDDGERAGAR PPTHDTNRSI VNSRIAPLVD
WWKTASALEQ HAMVKALSNA EVDDFDSPEG AKVQAFFADL DDDVHAKLDS LHLPVGRAAY
SEDTLVRLTR RMLSDGVDLY TARLQEFGIE PSWTPPTPRI GEPVGNPAVD RVLKTVSRWL
ESATKTWGAP ERVIIEHVRE GFVTEKRARE MDGDMRRRAA RNAKLFQEMQ EKLNVQGKPS
RADLWRYQSV QRQNCQCAYC GSPITFSNSE MDHIVPRAGQ GSTNTRENLV AVCHRCNQSK
GNTPFAIWAK NTSIEGVSVK EAVERTRHWV TDTGMRSTDF KKFTKAVVER FQRATMDEEI
DARSMESVAW MANELRSRVA QHFASHGTTV RVYRGSLTAE ARRASGISGK LKFFDGVGKS
RLDRRHHAID AAVIAFTSDY VAETLAVRSN LKQSQAHRQE APQWREFTGK DAEHRAAWRV
WCQKMEKLSA LLTEDLRDDR VVVMSNVRLR LGNGSAHKET IGKLSKVKLS SQLSVSDIDK
ASSEALWCAL TREPGFDPKE GLPANPERHI RVNGTHVYAG DNIGLFPVSA GSIALRGGYA
ELGSSFHHAR VYKITSGKKP AFAMLRVYTI DLLPYRNQDL FSVELKPQTM SMRQAEKKLR
DALATGNAEY LGWLVVDDEL VVDTSKIATD QVKAVEAELG TIRRWRVDGF FSPSKLRLRP
LQMSKEGIKK ESAPELSKII DRPGWLPAVN KLFSDGNVTV VRRDSLGRVR LESTAHLPVT
WKVQ
