CAS9_NEIMA
ID CAS9_NEIMA Reviewed; 1082 AA.
AC A1IQ68;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 06-FEB-2007, sequence version 1.
DT 03-AUG-2022, entry version 74.
DE RecName: Full=CRISPR-associated endonuclease Cas9 {ECO:0000255|HAMAP-Rule:MF_01480};
DE EC=3.1.-.- {ECO:0000255|HAMAP-Rule:MF_01480};
GN Name=cas9 {ECO:0000255|HAMAP-Rule:MF_01480}; OrderedLocusNames=NMA0631;
OS Neisseria meningitidis serogroup A / serotype 4A (strain DSM 15465 /
OS Z2491).
OC Bacteria; Proteobacteria; Betaproteobacteria; Neisseriales; Neisseriaceae;
OC Neisseria.
OX NCBI_TaxID=122587;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DSM 15465 / Z2491;
RX PubMed=10761919; DOI=10.1038/35006655;
RA Parkhill J., Achtman M., James K.D., Bentley S.D., Churcher C.M.,
RA Klee S.R., Morelli G., Basham D., Brown D., Chillingworth T., Davies R.M.,
RA Davis P., Devlin K., Feltwell T., Hamlin N., Holroyd S., Jagels K.,
RA Leather S., Moule S., Mungall K.L., Quail M.A., Rajandream M.A.,
RA Rutherford K.M., Simmonds M., Skelton J., Whitehead S., Spratt B.G.,
RA Barrell B.G.;
RT "Complete DNA sequence of a serogroup A strain of Neisseria meningitidis
RT Z2491.";
RL Nature 404:502-506(2000).
RN [2]
RP FUNCTION AS AN ENDONUCLEASE, AND POSSIBLE BIOTECHNOLOGY.
RC STRAIN=DSM 15465 / Z2491;
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 C.jejuni strain
CC NCTC 11168 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; AL157959; CAM07896.1; -; Genomic_DNA.
DR PIR; H81982; H81982.
DR RefSeq; WP_002235162.1; NC_003116.1.
DR PDB; 6JHW; X-ray; 2.04 A; B/D=518-655.
DR PDBsum; 6JHW; -.
DR AlphaFoldDB; A1IQ68; -.
DR SMR; A1IQ68; -.
DR EnsemblBacteria; CAM07896; CAM07896; NMA0631.
DR KEGG; nma:NMA0631; -.
DR HOGENOM; CLU_007514_0_0_4; -.
DR OMA; GQGHMET; -.
DR BioCyc; NMEN122587:NMA_RS03200-MON; -.
DR Proteomes; UP000000626; 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; 1.
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; RNA-binding.
FT CHAIN 1..1082
FT /note="CRISPR-associated endonuclease Cas9"
FT /id="PRO_0000429985"
FT DOMAIN 512..667
FT /note="HNH Cas9-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01085"
FT ACT_SITE 16
FT /note="For RuvC-like nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT ACT_SITE 588
FT /note="Proton acceptor for HNH nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 16
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 16
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 504
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 508
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 508
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 723
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT HELIX 518..540
FT /evidence="ECO:0007829|PDB:6JHW"
FT HELIX 550..561
FT /evidence="ECO:0007829|PDB:6JHW"
FT TURN 562..564
FT /evidence="ECO:0007829|PDB:6JHW"
FT TURN 567..569
FT /evidence="ECO:0007829|PDB:6JHW"
FT HELIX 575..577
FT /evidence="ECO:0007829|PDB:6JHW"
FT TURN 581..583
FT /evidence="ECO:0007829|PDB:6JHW"
FT STRAND 584..590
FT /evidence="ECO:0007829|PDB:6JHW"
FT STRAND 596..599
FT /evidence="ECO:0007829|PDB:6JHW"
FT STRAND 603..607
FT /evidence="ECO:0007829|PDB:6JHW"
FT HELIX 608..610
FT /evidence="ECO:0007829|PDB:6JHW"
FT HELIX 619..622
FT /evidence="ECO:0007829|PDB:6JHW"
FT TURN 623..628
FT /evidence="ECO:0007829|PDB:6JHW"
FT HELIX 630..641
FT /evidence="ECO:0007829|PDB:6JHW"
FT HELIX 646..652
FT /evidence="ECO:0007829|PDB:6JHW"
SQ SEQUENCE 1082 AA; 124321 MW; 9472BB28F7739680 CRC64;
MAAFKPNPIN YILGLDIGIA SVGWAMVEID EDENPICLID LGVRVFERAE VPKTGDSLAM
ARRLARSVRR LTRRRAHRLL RARRLLKREG VLQAADFDEN GLIKSLPNTP WQLRAAALDR
KLTPLEWSAV LLHLIKHRGY LSQRKNEGET ADKELGALLK GVADNAHALQ TGDFRTPAEL
ALNKFEKESG HIRNQRGDYS HTFSRKDLQA ELILLFEKQK EFGNPHVSGG LKEGIETLLM
TQRPALSGDA VQKMLGHCTF EPAEPKAAKN TYTAERFIWL TKLNNLRILE QGSERPLTDT
ERATLMDEPY RKSKLTYAQA RKLLGLEDTA FFKGLRYGKD NAEASTLMEM KAYHAISRAL
EKEGLKDKKS PLNLSPELQD EIGTAFSLFK TDEDITGRLK DRIQPEILEA LLKHISFDKF
VQISLKALRR IVPLMEQGKR YDEACAEIYG DHYGKKNTEE KIYLPPIPAD EIRNPVVLRA
LSQARKVING VVRRYGSPAR IHIETAREVG KSFKDRKEIE KRQEENRKDR EKAAAKFREY
FPNFVGEPKS KDILKLRLYE QQHGKCLYSG KEINLGRLNE KGYVEIDHAL PFSRTWDDSF
NNKVLVLGSE NQNKGNQTPY EYFNGKDNSR EWQEFKARVE TSRFPRSKKQ RILLQKFDED
GFKERNLNDT RYVNRFLCQF VADRMRLTGK GKKRVFASNG QITNLLRGFW GLRKVRAEND
RHHALDAVVV ACSTVAMQQK ITRFVRYKEM NAFDGKTIDK ETGEVLHQKT HFPQPWEFFA
QEVMIRVFGK PDGKPEFEEA DTPEKLRTLL AEKLSSRPEA VHEYVTPLFV SRAPNRKMSG
QGHMETVKSA KRLDEGVSVL RVPLTQLKLK DLEKMVNRER EPKLYEALKA RLEAHKDDPA
KAFAEPFYKY DKAGNRTQQV KAVRVEQVQK TGVWVRNHNG IADNATMVRV DVFEKGDKYY
LVPIYSWQVA KGILPDRAVV QGKDEEDWQL IDDSFNFKFS LHPNDLVEVI TKKARMFGYF
ASCHRGTGNI NIRIHDLDHK IGKNGILEGI GVKTALSFQK YQIDELGKEI RPCRLKKRPP
VR
