CAS9_STRMU
ID CAS9_STRMU Reviewed; 1345 AA.
AC Q8DTE3;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2003, sequence version 1.
DT 03-AUG-2022, entry version 93.
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}; Synonyms=csn1;
GN OrderedLocusNames=SMU_1405c;
OS Streptococcus mutans serotype c (strain ATCC 700610 / UA159).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Streptococcus.
OX NCBI_TaxID=210007;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 700610 / UA159;
RX PubMed=12397186; DOI=10.1073/pnas.172501299;
RA Ajdic D.J., McShan W.M., McLaughlin R.E., Savic G., Chang J., Carson M.B.,
RA Primeaux C., Tian R., Kenton S., Jia H.G., Lin S.P., Qian Y., Li S.,
RA Zhu H., Najar F.Z., Lai H., White J., Roe B.A., Ferretti J.J.;
RT "Genome sequence of Streptococcus mutans UA159, a cariogenic dental
RT pathogen.";
RL Proc. Natl. Acad. Sci. U.S.A. 99:14434-14439(2002).
RN [2]
RP FUNCTION AS AN ENDONUCLEASE, FUNCTION IN GUIDE RNA PROCESSING, AND POSSIBLE
RP BIOTECHNOLOGY.
RC STRAIN=ATCC 700610 / UA159;
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 Complements the gRNA coprocessing defect in a cas9 deletion in
CC S.pyogenes strain 370 and cuts target plasmid in Cas9:gRNAs mixing
CC experiments with S.thermophilus CRISPR3 from strain LMD-9.
CC {ECO:0000255|HAMAP-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-A 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; AE014133; AAN59070.1; -; Genomic_DNA.
DR RefSeq; NP_721764.1; NC_004350.2.
DR RefSeq; WP_002263549.1; NC_004350.2.
DR AlphaFoldDB; Q8DTE3; -.
DR SMR; Q8DTE3; -.
DR STRING; 210007.SMU_1405c; -.
DR PRIDE; Q8DTE3; -.
DR EnsemblBacteria; AAN59070; AAN59070; SMU_1405c.
DR KEGG; smu:SMU_1405c; -.
DR PATRIC; fig|210007.7.peg.1250; -.
DR eggNOG; COG3513; Bacteria.
DR HOGENOM; CLU_005604_0_0_9; -.
DR OMA; TDRHSIK; -.
DR Proteomes; UP000002512; 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; -; 1.
DR HAMAP; MF_01480; Cas9; 1.
DR InterPro; IPR028629; Cas9.
DR InterPro; IPR032239; Cas9-BH.
DR InterPro; IPR032237; Cas9_PI.
DR InterPro; IPR032240; Cas9_REC.
DR InterPro; IPR033114; HNH_CAS9.
DR InterPro; IPR003615; HNH_nuc.
DR InterPro; IPR036397; RNaseH_sf.
DR Pfam; PF16593; Cas9-BH; 1.
DR Pfam; PF16595; Cas9_PI; 1.
DR Pfam; PF16592; Cas9_REC; 1.
DR Pfam; PF13395; HNH_4; 1.
DR TIGRFAMs; TIGR01865; cas_Csn1; 1.
DR PROSITE; PS51749; HNH_CAS9; 1.
PE 1: Evidence at protein level;
KW Antiviral defense; DNA-binding; Endonuclease; Hydrolase; Magnesium;
KW Manganese; Metal-binding; Nuclease; Reference proteome; RNA-binding.
FT CHAIN 1..1345
FT /note="CRISPR-associated endonuclease Cas9"
FT /id="PRO_0000429988"
FT DOMAIN 770..921
FT /note="HNH Cas9-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01085"
FT ACT_SITE 10
FT /note="For RuvC-like nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT ACT_SITE 840
FT /note="Proton acceptor for HNH nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 10
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 10
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 762
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 766
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 766
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 983
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
SQ SEQUENCE 1345 AA; 156624 MW; DE459510910B30D9 CRC64;
MKKPYSIGLD IGTNSVGWAV VTDDYKVPAK KMKVLGNTDK SHIEKNLLGA LLFDSGNTAE
DRRLKRTARR RYTRRRNRIL YLQEIFSEEM GKVDDSFFHR LEDSFLVTED KRGERHPIFG
NLEEEVKYHE NFPTIYHLRQ YLADNPEKVD LRLVYLALAH IIKFRGHFLI EGKFDTRNND
VQRLFQEFLA VYDNTFENSS LQEQNVQVEE ILTDKISKSA KKDRVLKLFP NEKSNGRFAE
FLKLIVGNQA DFKKHFELEE KAPLQFSKDT YEEELEVLLA QIGDNYAELF LSAKKLYDSI
LLSGILTVTD VGTKAPLSAS MIQRYNEHQM DLAQLKQFIR QKLSDKYNEV FSDVSKDGYA
GYIDGKTNQE AFYKYLKGLL NKIEGSGYFL DKIEREDFLR KQRTFDNGSI PHQIHLQEMR
AIIRRQAEFY PFLADNQDRI EKLLTFRIPY YVGPLARGKS DFAWLSRKSA DKITPWNFDE
IVDKESSAEA FINRMTNYDL YLPNQKVLPK HSLLYEKFTV YNELTKVKYK TEQGKTAFFD
ANMKQEIFDG VFKVYRKVTK DKLMDFLEKE FDEFRIVDLT GLDKENKVFN ASYGTYHDLC
KILDKDFLDN SKNEKILEDI VLTLTLFEDR EMIRKRLENY SDLLTKEQVK KLERRHYTGW
GRLSAELIHG IRNKESRKTI LDYLIDDGNS NRNFMQLIND DALSFKEEIA KAQVIGETDN
LNQVVSDIAG SPAIKKGILQ SLKIVDELVK IMGHQPENIV VEMARENQFT NQGRRNSQQR
LKGLTDSIKE FGSQILKEHP VENSQLQNDR LFLYYLQNGR DMYTGEELDI DYLSQYDIDH
IIPQAFIKDN SIDNRVLTSS KENRGKSDDV PSKDVVRKMK SYWSKLLSAK LITQRKFDNL
TKAERGGLTD DDKAGFIKRQ LVETRQITKH VARILDERFN TETDENNKKI RQVKIVTLKS
NLVSNFRKEF ELYKVREIND YHHAHDAYLN AVIGKALLGV YPQLEPEFVY GDYPHFHGHK
ENKATAKKFF YSNIMNFFKK DDVRTDKNGE IIWKKDEHIS NIKKVLSYPQ VNIVKKVEEQ
TGGFSKESIL PKGNSDKLIP RKTKKFYWDT KKYGGFDSPI VAYSILVIAD IEKGKSKKLK
TVKALVGVTI MEKMTFERDP VAFLERKGYR NVQEENIIKL PKYSLFKLEN GRKRLLASAR
ELQKGNEIVL PNHLGTLLYH AKNIHKVDEP KHLDYVDKHK DEFKELLDVV SNFSKKYTLA
EGNLEKIKEL YAQNNGEDLK ELASSFINLL TFTAIGAPAT FKFFDKNIDR KRYTSTTEIL
NATLIHQSIT GLYETRIDLN KLGGD
