CAS9A_STRTD
ID CAS9A_STRTD Reviewed; 1121 AA.
AC Q03LF7;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 14-NOV-2006, sequence version 1.
DT 03-AUG-2022, entry version 81.
DE RecName: Full=CRISPR-associated endonuclease Cas9 1 {ECO:0000255|HAMAP-Rule:MF_01480};
DE EC=3.1.-.- {ECO:0000255|HAMAP-Rule:MF_01480};
DE AltName: Full=Cas9**;
GN Name=cas9-1 {ECO:0000255|HAMAP-Rule:MF_01480}; Synonyms=csn1;
GN OrderedLocusNames=STER_0709;
OS Streptococcus thermophilus (strain ATCC BAA-491 / LMD-9).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Streptococcus.
OX NCBI_TaxID=322159;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC BAA-491 / LMD-9;
RX PubMed=17030793; DOI=10.1073/pnas.0607117103;
RA Makarova K.S., Slesarev A., Wolf Y.I., Sorokin A., Mirkin B., Koonin E.V.,
RA Pavlov A., Pavlova N., Karamychev V., Polouchine N., Shakhova V.,
RA Grigoriev I., Lou Y., Rohksar D., Lucas S., Huang K., Goodstein D.M.,
RA Hawkins T., Plengvidhya V., Welker D., Hughes J., Goh Y., Benson A.,
RA Baldwin K., Lee J.-H., Diaz-Muniz I., Dosti B., Smeianov V., Wechter W.,
RA Barabote R., Lorca G., Altermann E., Barrangou R., Ganesan B., Xie Y.,
RA Rawsthorne H., Tamir D., Parker C., Breidt F., Broadbent J.R., Hutkins R.,
RA O'Sullivan D., Steele J., Unlu G., Saier M.H. Jr., Klaenhammer T.,
RA Richardson P., Kozyavkin S., Weimer B.C., Mills D.A.;
RT "Comparative genomics of the lactic acid bacteria.";
RL Proc. Natl. Acad. Sci. U.S.A. 103:15611-15616(2006).
RN [2]
RP FUNCTION AS AN ENDONUCLEASE, AND POSSIBLE BIOTECHNOLOGY.
RC STRAIN=ATCC BAA-491 / LMD-9;
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 when Cas9 and gRNAs are mixed. {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 (Probable). {ECO:0000305}.
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}.
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DR EMBL; CP000419; ABJ65965.1; -; Genomic_DNA.
DR RefSeq; WP_011680957.1; NC_008532.1.
DR PDB; 6M0V; X-ray; 3.00 A; A=2-1121.
DR PDB; 6M0W; X-ray; 2.76 A; A=2-1121.
DR PDB; 6M0X; X-ray; 2.56 A; A=2-1121.
DR PDB; 6RJ9; EM; 3.20 A; C=1-1121.
DR PDB; 6RJA; EM; 3.00 A; C/F=1-1121.
DR PDB; 6RJD; EM; 3.30 A; C=1-1121.
DR PDB; 6RJG; EM; 3.20 A; C=1-1121.
DR PDBsum; 6M0V; -.
DR PDBsum; 6M0W; -.
DR PDBsum; 6M0X; -.
DR PDBsum; 6RJ9; -.
DR PDBsum; 6RJA; -.
DR PDBsum; 6RJD; -.
DR PDBsum; 6RJG; -.
DR AlphaFoldDB; Q03LF7; -.
DR SMR; Q03LF7; -.
DR PRIDE; Q03LF7; -.
DR KEGG; ste:STER_0709; -.
DR HOGENOM; CLU_007514_0_0_9; -.
DR OMA; GQGHMET; -.
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; -; 2.
DR HAMAP; MF_01480; Cas9; 1.
DR InterPro; IPR028629; Cas9.
DR InterPro; IPR040619; Cas9_alpha-helical_lobe.
DR InterPro; IPR040555; Cas9_PI2.
DR InterPro; IPR040656; Cas9_WED_dom.
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; PF18070; Cas9_PI2; 1.
DR Pfam; PF18061; CRISPR_Cas9_WED; 1.
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..1121
FT /note="CRISPR-associated endonuclease Cas9 1"
FT /id="PRO_0000429989"
FT DOMAIN 516..684
FT /note="HNH Cas9-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01085"
FT ACT_SITE 9
FT /note="For RuvC-like nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT ACT_SITE 599
FT /note="Proton acceptor for HNH nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 9
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 9
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 509
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 513
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 513
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 738
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT STRAND 5..10
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 12..20
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 22..24
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 27..34
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 38..41
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 43..72
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 79..81
FT /evidence="ECO:0007829|PDB:6M0V"
FT STRAND 84..86
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 88..94
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 95..97
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 102..113
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 134..146
FT /evidence="ECO:0007829|PDB:6RJA"
FT HELIX 151..162
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 166..173
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 176..181
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 186..203
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 209..220
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 225..227
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 232..234
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 239..241
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 243..245
FT /evidence="ECO:0007829|PDB:6RJD"
FT HELIX 253..255
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 260..262
FT /evidence="ECO:0007829|PDB:6M0W"
FT HELIX 273..285
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 290..295
FT /evidence="ECO:0007829|PDB:6M0V"
FT HELIX 298..310
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 316..325
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 331..333
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 341..343
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 351..357
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 360..362
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 364..366
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 369..381
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 385..395
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 403..415
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 417..420
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 424..427
FT /evidence="ECO:0007829|PDB:6RJA"
FT HELIX 429..433
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 436..441
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 446..453
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 465..467
FT /evidence="ECO:0007829|PDB:6RJG"
FT HELIX 470..473
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 474..476
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 480..500
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 504..510
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 517..545
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 547..550
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 553..556
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 562..571
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 572..574
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 577..579
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 585..590
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 592..594
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 595..601
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 603..605
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 611..613
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 614..618
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 619..625
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 630..633
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 634..636
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 638..640
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 643..652
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 654..656
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 658..665
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 681..704
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 710..715
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 716..725
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 732..734
FT /evidence="ECO:0007829|PDB:6M0V"
FT HELIX 736..749
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 750..752
FT /evidence="ECO:0007829|PDB:6M0W"
FT HELIX 792..800
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 805..807
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 809..812
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 828..833
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 842..850
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 852..854
FT /evidence="ECO:0007829|PDB:6RJA"
FT HELIX 855..868
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 869..871
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 873..877
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 879..884
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 886..892
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 895..898
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 904..906
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 909..917
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 925..927
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 935..941
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 944..947
FT /evidence="ECO:0007829|PDB:6RJG"
FT STRAND 957..959
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 964..972
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 973..976
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 977..984
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 985..987
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 992..994
FT /evidence="ECO:0007829|PDB:6M0X"
FT HELIX 1001..1010
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 1018..1025
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 1028..1033
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 1034..1036
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 1039..1047
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 1050..1052
FT /evidence="ECO:0007829|PDB:6M0V"
FT STRAND 1055..1059
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 1061..1065
FT /evidence="ECO:0007829|PDB:6M0X"
FT TURN 1074..1076
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 1081..1083
FT /evidence="ECO:0007829|PDB:6RJA"
FT STRAND 1085..1088
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 1094..1101
FT /evidence="ECO:0007829|PDB:6M0X"
FT STRAND 1107..1111
FT /evidence="ECO:0007829|PDB:6M0X"
SQ SEQUENCE 1121 AA; 129447 MW; F67743AF92C43F06 CRC64;
MSDLVLGLDI GIGSVGVGIL NKVTGEIIHK NSRIFPAAQA ENNLVRRTNR QGRRLARRKK
HRRVRLNRLF EESGLITDFT KISINLNPYQ LRVKGLTDEL SNEELFIALK NMVKHRGISY
LDDASDDGNS SVGDYAQIVK ENSKQLETKT PGQIQLERYQ TYGQLRGDFT VEKDGKKHRL
INVFPTSAYR SEALRILQTQ QEFNPQITDE FINRYLEILT GKRKYYHGPG NEKSRTDYGR
YRTSGETLDN IFGILIGKCT FYPDEFRAAK ASYTAQEFNL LNDLNNLTVP TETKKLSKEQ
KNQIINYVKN EKAMGPAKLF KYIAKLLSCD VADIKGYRID KSGKAEIHTF EAYRKMKTLE
TLDIEQMDRE TLDKLAYVLT LNTEREGIQE ALEHEFADGS FSQKQVDELV QFRKANSSIF
GKGWHNFSVK LMMELIPELY ETSEEQMTIL TRLGKQKTTS SSNKTKYIDE KLLTEEIYNP
VVAKSVRQAI KIVNAAIKEY GDFDNIVIEM ARETNEDDEK KAIQKIQKAN KDEKDAAMLK
AANQYNGKAE LPHSVFHGHK QLATKIRLWH QQGERCLYTG KTISIHDLIN NSNQFEVDHI
LPLSITFDDS LANKVLVYAT ANQEKGQRTP YQALDSMDDA WSFRELKAFV RESKTLSNKK
KEYLLTEEDI SKFDVRKKFI ERNLVDTRYA SRVVLNALQE HFRAHKIDTK VSVVRGQFTS
QLRRHWGIEK TRDTYHHHAV DALIIAASSQ LNLWKKQKNT LVSYSEDQLL DIETGELISD
DEYKESVFKA PYQHFVDTLK SKEFEDSILF SYQVDSKFNR KISDATIYAT RQAKVGKDKA
DETYVLGKIK DIYTQDGYDA FMKIYKKDKS KFLMYRHDPQ TFEKVIEPIL ENYPNKQINE
KGKEVPCNPF LKYKEEHGYI RKYSKKGNGP EIKSLKYYDS KLGNHIDITP KDSNNKVVLQ
SVSPWRADVY FNKTTGKYEI LGLKYADLQF EKGTGTYKIS QEKYNDIKKK EGVDSDSEFK
FTLYKNDLLL VKDTETKEQQ LFRFLSRTMP KQKHYVELKP YDKQKFEGGE ALIKVLGNVA
NSGQCKKGLG KSNISIYKVR TDVLGNQHII KNEGDKPKLD F
