CAS9_PASMU
ID CAS9_PASMU Reviewed; 1056 AA.
AC Q9CLT2;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2001, sequence version 1.
DT 03-AUG-2022, entry version 102.
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=PM1127;
OS Pasteurella multocida (strain Pm70).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pasteurellales;
OC Pasteurellaceae; Pasteurella.
OX NCBI_TaxID=272843;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=Pm70;
RX PubMed=11248100; DOI=10.1073/pnas.051634598;
RA May B.J., Zhang Q., Li L.L., Paustian M.L., Whittam T.S., Kapur V.;
RT "Complete genomic sequence of Pasteurella multocida Pm70.";
RL Proc. Natl. Acad. Sci. U.S.A. 98:3460-3465(2001).
RN [2]
RP FUNCTION AS AN ENDONUCLEASE, AND POSSIBLE BIOTECHNOLOGY.
RC STRAIN=Pm70;
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 N.meningitidis strain Z2491. {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:0000305}.
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DR EMBL; AE004439; AAK03211.1; -; Genomic_DNA.
DR RefSeq; WP_010907033.1; NC_002663.1.
DR AlphaFoldDB; Q9CLT2; -.
DR SMR; Q9CLT2; -.
DR STRING; 272843.PM1127; -.
DR EnsemblBacteria; AAK03211; AAK03211; PM1127.
DR KEGG; pmu:PM1127; -.
DR PATRIC; fig|272843.6.peg.1138; -.
DR HOGENOM; CLU_007514_0_0_6; -.
DR OMA; GQGHMET; -.
DR Proteomes; UP000000809; 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; IPR040619; Cas9_alpha-helical_lobe.
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; 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 Antiviral defense; DNA-binding; Endonuclease; Hydrolase; Magnesium;
KW Manganese; Metal-binding; Nuclease; Reference proteome; RNA-binding.
FT CHAIN 1..1056
FT /note="CRISPR-associated endonuclease Cas9"
FT /id="PRO_0000429987"
FT DOMAIN 504..659
FT /note="HNH Cas9-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01085"
FT ACT_SITE 13
FT /note="For RuvC-like nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT ACT_SITE 580
FT /note="Proton acceptor for HNH nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 13
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 13
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 496
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 500
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 500
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 715
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
SQ SEQUENCE 1056 AA; 121836 MW; 1EB44187E232215E CRC64;
MQTTNLSYIL GLDLGIASVG WAVVEINENE DPIGLIDVGV RIFERAEVPK TGESLALSRR
LARSTRRLIR RRAHRLLLAK RFLKREGILS TIDLEKGLPN QAWELRVAGL ERRLSAIEWG
AVLLHLIKHR GYLSKRKNES QTNNKELGAL LSGVAQNHQL LQSDDYRTPA ELALKKFAKE
EGHIRNQRGA YTHTFNRLDL LAELNLLFAQ QHQFGNPHCK EHIQQYMTEL LMWQKPALSG
EAILKMLGKC THEKNEFKAA KHTYSAERFV WLTKLNNLRI LEDGAERALN EEERQLLINH
PYEKSKLTYA QVRKLLGLSE QAIFKHLRYS KENAESATFM ELKAWHAIRK ALENQGLKDT
WQDLAKKPDL LDEIGTAFSL YKTDEDIQQY LTNKVPNSVI NALLVSLNFD KFIELSLKSL
RKILPLMEQG KRYDQACREI YGHHYGEANQ KTSQLLPAIP AQEIRNPVVL RTLSQARKVI
NAIIRQYGSP ARVHIETGRE LGKSFKERRE IQKQQEDNRT KRESAVQKFK ELFSDFSSEP
KSKDILKFRL YEQQHGKCLY SGKEINIHRL NEKGYVEIDH ALPFSRTWDD SFNNKVLVLA
SENQNKGNQT PYEWLQGKIN SERWKNFVAL VLGSQCSAAK KQRLLTQVID DNKFIDRNLN
DTRYIARFLS NYIQENLLLV GKNKKNVFTP NGQITALLRS RWGLIKAREN NNRHHALDAI
VVACATPSMQ QKITRFIRFK EVHPYKIENR YEMVDQESGE IISPHFPEPW AYFRQEVNIR
VFDNHPDTVL KEMLPDRPQA NHQFVQPLFV SRAPTRKMSG QGHMETIKSA KRLAEGISVL
RIPLTQLKPN LLENMVNKER EPALYAGLKA RLAEFNQDPA KAFATPFYKQ GGQQVKAIRV
EQVQKSGVLV RENNGVADNA SIVRTDVFIK NNKFFLVPIY TWQVAKGILP NKAIVAHKNE
DEWEEMDEGA KFKFSLFPND LVELKTKKEY FFGYYIGLDR ATGNISLKEH DGEISKGKDG
VYRVGVKLAL SFEKYQVDEL GKNRQICRPQ QRQPVR
