ID CAS9_LISIN Reviewed; 1334 AA. AC Q927P4; DT 06-MAR-2013, integrated into UniProtKB/Swiss-Prot. DT 01-DEC-2001, sequence version 1. DT 03-AUG-2022, entry version 103. 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=lin2744; OS Listeria innocua serovar 6a (strain ATCC BAA-680 / CLIP 11262). OC Bacteria; Firmicutes; Bacilli; Bacillales; Listeriaceae; Listeria. OX NCBI_TaxID=272626; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=ATCC BAA-680 / CLIP 11262; RX PubMed=11679669; DOI=10.1126/science.1063447; RA Glaser P., Frangeul L., Buchrieser C., Rusniok C., Amend A., Baquero F., RA Berche P., Bloecker H., Brandt P., Chakraborty T., Charbit A., RA Chetouani F., Couve E., de Daruvar A., Dehoux P., Domann E., RA Dominguez-Bernal G., Duchaud E., Durant L., Dussurget O., Entian K.-D., RA Fsihi H., Garcia-del Portillo F., Garrido P., Gautier L., Goebel W., RA Gomez-Lopez N., Hain T., Hauf J., Jackson D., Jones L.-M., Kaerst U., RA Kreft J., Kuhn M., Kunst F., Kurapkat G., Madueno E., Maitournam A., RA Mata Vicente J., Ng E., Nedjari H., Nordsiek G., Novella S., de Pablos B., RA Perez-Diaz J.-C., Purcell R., Remmel B., Rose M., Schlueter T., Simoes N., RA Tierrez A., Vazquez-Boland J.-A., Voss H., Wehland J., Cossart P.; RT "Comparative genomics of Listeria species."; RL Science 294:849-852(2001). RN [2] RP FUNCTION AS AN DNA ENDONUCLEASE, SUBUNIT, POSSIBLE BIOTECHNOLOGY, AND RP RNA-BINDING. RC STRAIN=ATCC BAA-680 / CLIP 11262; RX PubMed=22745249; DOI=10.1126/science.1225829; RA Jinek M., Chylinski K., Fonfara I., Hauer M., Doudna J.A., Charpentier E.; RT "A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial RT immunity."; RL Science 337:816-821(2012). 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. CC {ECO:0000255|HAMAP-Rule:MF_01480, ECO:0000269|PubMed:22745249}. CC -!- COFACTOR: CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000305}; CC Note=Endonuclease activity on target dsDNA requires Mg(2+). CC {ECO:0000305}; CC -!- SUBUNIT: Monomer (By similarity). Binds crRNA and tracrRNA. CC {ECO:0000255|HAMAP-Rule:MF_01480, ECO:0000269|PubMed:22745249}. 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; AL596173; CAC97970.1; -; Genomic_DNA. DR PIR; AB1775; AB1775. DR RefSeq; WP_010991369.1; NC_003212.1. DR AlphaFoldDB; Q927P4; -. DR SMR; Q927P4; -. DR STRING; 272626.lin2744; -. DR EnsemblBacteria; CAC97970; CAC97970; CAC97970. DR KEGG; lin:lin2744; -. DR eggNOG; COG3513; Bacteria. DR HOGENOM; CLU_005604_0_0_9; -. DR OMA; TDRHSIK; -. DR OrthoDB; 27691at2; -. DR Proteomes; UP000002513; 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; RNA-binding. FT CHAIN 1..1334 FT /note="CRISPR-associated endonuclease Cas9" FT /id="PRO_0000421685" FT DOMAIN 773..924 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 843 FT /note="Proton acceptor for HNH nuclease domain" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480" FT BINDING 10 FT /ligand="Mn(2+)" FT /ligand_id="ChEBI:CHEBI:29035" FT /ligand_label="1" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480" FT BINDING 10 FT /ligand="Mn(2+)" FT /ligand_id="ChEBI:CHEBI:29035" FT /ligand_label="2" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480" FT BINDING 765 FT /ligand="Mn(2+)" FT /ligand_id="ChEBI:CHEBI:29035" FT /ligand_label="1" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480" FT BINDING 769 FT /ligand="Mn(2+)" FT /ligand_id="ChEBI:CHEBI:29035" FT /ligand_label="1" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480" FT BINDING 769 FT /ligand="Mn(2+)" FT /ligand_id="ChEBI:CHEBI:29035" FT /ligand_label="2" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480" FT BINDING 986 FT /ligand="Mn(2+)" FT /ligand_id="ChEBI:CHEBI:29035" FT /ligand_label="2" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480" SQ SEQUENCE 1334 AA; 154807 MW; 95557D51A4B7185E CRC64; MKKPYTIGLD IGTNSVGWAV LTDQYDLVKR KMKIAGDSEK KQIKKNFWGV RLFDEGQTAA DRRMARTARR RIERRRNRIS YLQGIFAEEM SKTDANFFCR LSDSFYVDNE KRNSRHPFFA TIEEEVEYHK NYPTIYHLRE ELVNSSEKAD LRLVYLALAH IIKYRGNFLI EGALDTQNTS VDGIYKQFIQ TYNQVFASGI EDGSLKKLED NKDVAKILVE KVTRKEKLER ILKLYPGEKS AGMFAQFISL IVGSKGNFQK PFDLIEKSDI ECAKDSYEED LESLLALIGD EYAELFVAAK NAYSAVVLSS IITVAETETN AKLSASMIER FDTHEEDLGE LKAFIKLHLP KHYEEIFSNT EKHGYAGYID GKTKQADFYK YMKMTLENIE GADYFIAKIE KENFLRKQRT FDNGAIPHQL HLEELEAILH QQAKYYPFLK ENYDKIKSLV TFRIPYFVGP LANGQSEFAW LTRKADGEIR PWNIEEKVDF GKSAVDFIEK MTNKDTYLPK ENVLPKHSLC YQKYLVYNEL TKVRYINDQG KTSYFSGQEK EQIFNDLFKQ KRKVKKKDLE LFLRNMSHVE SPTIEGLEDS FNSSYSTYHD LLKVGIKQEI LDNPVNTEML ENIVKILTVF EDKRMIKEQL QQFSDVLDGV VLKKLERRHY TGWGRLSAKL LMGIRDKQSH LTILDYLMND DGLNRNLMQL INDSNLSFKS IIEKEQVTTA DKDIQSIVAD LAGSPAIKKG ILQSLKIVDE LVSVMGYPPQ TIVVEMAREN QTTGKGKNNS RPRYKSLEKA IKEFGSQILK EHPTDNQELR NNRLYLYYLQ NGKDMYTGQD LDIHNLSNYD IDHIVPQSFI TDNSIDNLVL TSSAGNREKG DDVPPLEIVR KRKVFWEKLY QGNLMSKRKF DYLTKAERGG LTEADKARFI HRQLVETRQI TKNVANILHQ RFNYEKDDHG NTMKQVRIVT LKSALVSQFR KQFQLYKVRD VNDYHHAHDA YLNGVVANTL LKVYPQLEPE FVYGDYHQFD WFKANKATAK KQFYTNIMLF FAQKDRIIDE NGEILWDKKY LDTVKKVMSY RQMNIVKKTE IQKGEFSKAT IKPKGNSSKL IPRKTNWDPM KYGGLDSPNM AYAVVIEYAK GKNKLVFEKK IIRVTIMERK AFEKDEKAFL EEQGYRQPKV LAKLPKYTLY ECEEGRRRML ASANEAQKGN QQVLPNHLVT LLHHAANCEV SDGKSLDYIE SNREMFAELL AHVSEFAKRY TLAEANLNKI NQLFEQNKEG DIKAIAQSFV DLMAFNAMGA PASFKFFETT IERKRYNNLK ELLNSTIIYQ SITGLYESRK RLDD //