CAS9_ACTNH
ID CAS9_ACTNH Reviewed; 1101 AA.
AC J3F2B0;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 03-OCT-2012, sequence version 1.
DT 03-AUG-2022, entry version 41.
DE RecName: Full=CRISPR-associated endonuclease Cas9;
DE EC=3.1.-.-;
DE AltName: Full=AnaCas9 {ECO:0000303|PubMed:24505130};
GN Name=cas9; ORFNames=HMPREF1129_2620;
OS Actinomyces naeslundii (strain ATCC 12104 / DSM 43013 / CCUG 2238 / JCM
OS 8349 / NCTC 10301 / Howell 279).
OC Bacteria; Actinobacteria; Actinomycetales; Actinomycetaceae; Actinomyces.
OX NCBI_TaxID=1115803;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 12104 / DSM 43013 / CCUG 2238 / JCM 8349 / NCTC 10301 / Howell
RC 279;
RA Durkin A.S., McCorrison J., Torralba M., Gillis M., Methe B., Sutton G.,
RA Nelson K.E.;
RL Submitted (JUL-2012) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH MAGNESIUM; MANGANESE
RP AND ZINC, FUNCTION, ACTIVE SITE, COFACTOR, DOMAIN, AND POSSIBLE
RP BIOTECHNOLOGY.
RC STRAIN=ATCC 12104 / DSM 43013 / CCUG 2238 / JCM 8349 / NCTC 10301 / Howell
RC 279;
RX PubMed=24505130; DOI=10.1126/science.1247997;
RA Jinek M., Jiang F., Taylor D.W., Sternberg S.H., Kaya E., Ma E., Anders C.,
RA Hauer M., Zhou K., Lin S., Kaplan M., Iavarone A.T., Charpentier E.,
RA Nogales E., Doudna J.A.;
RT "Structures of Cas9 endonucleases reveal RNA-mediated conformational
RT activation.";
RL Science 343:1247997-1247997(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 {ECO:0000250, ECO:0000269|PubMed:24505130}.
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:24505130};
CC Note=Binds 2 Mg(2+) per subunit. {ECO:0000269|PubMed:24505130};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:24505130};
CC Note=Binds 2 Mn(2+) per subunit. {ECO:0000269|PubMed:24505130};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:24505130};
CC Note=Binds 1 Zn(2+) per subunit, which may stabilize the HNH Cas9
CC architecture. {ECO:0000269|PubMed:24505130};
CC -!- SUBUNIT: Monomer. Binds crRNA and tracrRNA (Probable).
CC {ECO:0000305|PubMed:24505130}.
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 (Probable).
CC {ECO:0000305|PubMed:24505130}.
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; ALJK01000149; EJN84392.1; -; Genomic_DNA.
DR PDB; 4OGC; X-ray; 2.80 A; A=1-1101.
DR PDB; 4OGE; X-ray; 2.20 A; A=1-1101.
DR PDBsum; 4OGC; -.
DR PDBsum; 4OGE; -.
DR AlphaFoldDB; J3F2B0; -.
DR SMR; J3F2B0; -.
DR STRING; 1115803.HMPREF1129_2620; -.
DR PRIDE; J3F2B0; -.
DR EnsemblBacteria; EJN84392; EJN84392; HMPREF1129_2620.
DR PATRIC; fig|1115803.3.peg.1655; -.
DR eggNOG; COG3513; Bacteria.
DR Proteomes; UP000007814; Unassembled WGS sequence.
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; RNA-binding; Zinc.
FT CHAIN 1..1101
FT /note="CRISPR-associated endonuclease Cas9"
FT /id="PRO_0000429982"
FT DOMAIN 513..675
FT /note="HNH Cas9-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01085"
FT REGION 1..64
FT /note="RuvC-I"
FT /evidence="ECO:0000305|PubMed:24505130"
FT REGION 64..468
FT /note="Recognition lobe"
FT /evidence="ECO:0000305|PubMed:24505130"
FT REGION 253..273
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 468..513
FT /note="RuvC-II"
FT /evidence="ECO:0000305|PubMed:24505130"
FT REGION 674..822
FT /note="RuvC-III"
FT /evidence="ECO:0000305|PubMed:24505130"
FT REGION 924..1101
FT /note="PAM-interacting domain (PI)"
FT /evidence="ECO:0000305|PubMed:24505130"
FT ACT_SITE 17
FT /note="For RuvC-like nuclease domain"
FT /evidence="ECO:0000250|UniProtKB:Q99ZW2"
FT ACT_SITE 582
FT /note="Proton acceptor for HNH nuclease domain"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 17
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 17
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 505
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 566
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 569
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 581
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 586
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 588
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 590
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 602
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 605
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 606
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:24505130"
FT BINDING 736
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:24505130"
FT STRAND 11..18
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 20..30
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 36..46
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 66..91
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 92..94
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 138..147
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 148..153
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 227..230
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 237..242
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 243..246
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 247..249
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 256..258
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 278..290
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 293..297
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 301..303
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 306..318
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 326..333
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 337..339
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 361..368
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 372..379
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 384..394
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 401..404
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 405..407
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 410..414
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 415..418
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 431..443
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 448..456
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 476..496
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 500..505
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 514..542
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 550..561
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 567..569
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 575..577
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 579..584
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 586..590
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 595..597
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 598..601
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 603..609
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 614..621
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 628..636
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 647..662
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 672..675
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 680..692
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 696..701
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 705..713
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 716..718
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 735..745
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 748..766
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 773..775
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 781..806
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 810..813
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 834..837
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 838..840
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 844..848
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 850..852
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 853..860
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 867..869
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 878..881
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 884..887
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 890..894
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 900..902
FT /evidence="ECO:0007829|PDB:4OGC"
FT STRAND 911..915
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 918..921
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 926..934
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 936..938
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 940..947
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 948..954
FT /evidence="ECO:0007829|PDB:4OGE"
FT TURN 959..961
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 969..972
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 976..983
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 987..993
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 998..1000
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 1009..1017
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 1023..1031
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 1034..1042
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 1044..1046
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 1052..1058
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 1063..1066
FT /evidence="ECO:0007829|PDB:4OGE"
FT HELIX 1067..1072
FT /evidence="ECO:0007829|PDB:4OGE"
FT STRAND 1076..1078
FT /evidence="ECO:0007829|PDB:4OGE"
SQ SEQUENCE 1101 AA; 123754 MW; D93767E60402ACC2 CRC64;
MWYASLMSAH HLRVGIDVGT HSVGLATLRV DDHGTPIELL SALSHIHDSG VGKEGKKDHD
TRKKLSGIAR RARRLLHHRR TQLQQLDEVL RDLGFPIPTP GEFLDLNEQT DPYRVWRVRA
RLVEEKLPEE LRGPAISMAV RHIARHRGWR NPYSKVESLL SPAEESPFMK ALRERILATT
GEVLDDGITP GQAMAQVALT HNISMRGPEG ILGKLHQSDN ANEIRKICAR QGVSPDVCKQ
LLRAVFKADS PRGSAVSRVA PDPLPGQGSF RRAPKCDPEF QRFRIISIVA NLRISETKGE
NRPLTADERR HVVTFLTEDS QADLTWVDVA EKLGVHRRDL RGTAVHTDDG ERSAARPPID
ATDRIMRQTK ISSLKTWWEE ADSEQRGAMI RYLYEDPTDS ECAEIIAELP EEDQAKLDSL
HLPAGRAAYS RESLTALSDH MLATTDDLHE ARKRLFGVDD SWAPPAEAIN APVGNPSVDR
TLKIVGRYLS AVESMWGTPE VIHVEHVRDG FTSERMADER DKANRRRYND NQEAMKKIQR
DYGKEGYISR GDIVRLDALE LQGCACLYCG TTIGYHTCQL DHIVPQAGPG SNNRRGNLVA
VCERCNRSKS NTPFAVWAQK CGIPHVGVKE AIGRVRGWRK QTPNTSSEDL TRLKKEVIAR
LRRTQEDPEI DERSMESVAW MANELHHRIA AAYPETTVMV YRGSITAAAR KAAGIDSRIN
LIGEKGRKDR IDRRHHAVDA SVVALMEASV AKTLAERSSL RGEQRLTGKE QTWKQYTGST
VGAREHFEMW RGHMLHLTEL FNERLAEDKV YVTQNIRLRL SDGNAHTVNP SKLVSHRLGD
GLTVQQIDRA CTPALWCALT REKDFDEKNG LPAREDRAIR VHGHEIKSSD YIQVFSKRKK
TDSDRDETPF GAIAVRGGFV EIGPSIHHAR IYRVEGKKPV YAMLRVFTHD LLSQRHGDLF
SAVIPPQSIS MRCAEPKLRK AITTGNATYL GWVVVGDELE INVDSFTKYA IGRFLEDFPN
TTRWRICGYD TNSKLTLKPI VLAAEGLENP SSAVNEIVEL KGWRVAINVL TKVHPTVVRR
DALGRPRYSS RSNLPTSWTI E
