CAS9_NEIM8
ID CAS9_NEIM8 Reviewed; 1082 AA.
AC C9X1G5;
DT 03-SEP-2014, integrated into UniProtKB/Swiss-Prot.
DT 24-NOV-2009, sequence version 1.
DT 03-AUG-2022, entry version 58.
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=NMV_1993;
OS Neisseria meningitidis serogroup C (strain 8013).
OC Bacteria; Proteobacteria; Betaproteobacteria; Neisseriales; Neisseriaceae;
OC Neisseria.
OX NCBI_TaxID=604162;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=8013;
RX PubMed=19818133; DOI=10.1186/gb-2009-10-10-r110;
RA Rusniok C., Vallenet D., Floquet S., Ewles H., Mouze-Soulama C., Brown D.,
RA Lajus A., Buchrieser C., Medigue C., Glaser P., Pelicic V.;
RT "NeMeSys: a biological resource for narrowing the gap between sequence and
RT function in the human pathogen Neisseria meningitidis.";
RL Genome Biol. 10:R110.1-R110.13(2009).
RN [2]
RP FUNCTION IN PLASMID RESISTANCE, DISRUPTION PHENOTYPE, AND MUTAGENESIS OF
RP ASP-16 AND HIS-588.
RC STRAIN=8013;
RX PubMed=23706818; DOI=10.1016/j.molcel.2013.05.001;
RA Zhang Y., Heidrich N., Ampattu B.J., Gunderson C.W., Seifert H.S.,
RA Schoen C., Vogel J., Sontheimer E.J.;
RT "Processing-independent CRISPR RNAs limit natural transformation in
RT Neisseria meningitidis.";
RL Mol. Cell 50:488-503(2013).
RN [3]
RP FUNCTION AS AN ENDONUCLEASE, AND BIOTECHNOLOGY IN HUMAN CELLS.
RC STRAIN=8013;
RX PubMed=23940360; DOI=10.1073/pnas.1313587110;
RA Hou Z., Zhang Y., Propson N.E., Howden S.E., Chu L.F., Sontheimer E.J.,
RA Thomson J.A.;
RT "Efficient genome engineering in human pluripotent stem cells using Cas9
RT from Neisseria meningitidis.";
RL Proc. Natl. Acad. Sci. U.S.A. 110:15644-15649(2013).
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, although RNase 3 is not required for 5'-processing of crRNA in
CC this strain. Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or
CC circular dsDNA target complementary to the spacer; Cas9 is inactive in
CC the absence of the 2 guide RNAs (gRNA, PubMed:23940360). Cas9
CC recognizes the protospacer adjacent motif (PAM) in the CRISPR repeat
CC sequences to help distinguish self versus nonself, as targets within
CC the bacterial CRISPR locus do not have PAMs. PAM recognition is also
CC required for catalytic activity. Plasmids containing sequences
CC homologous to endogenous spacer elements and that have flanking PAM
CC consensus sequences cannot transform this strain unless the cas9 gene
CC is disrupted or critical residues of Cas9 are mutated.
CC {ECO:0000269|PubMed:23706818, ECO:0000269|PubMed:23940360}.
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 -!- DISRUPTION PHENOTYPE: Almost no mature crRNA or tracrRNA produced. Loss
CC of CRISPR interference during plasmid transformation.
CC {ECO:0000269|PubMed:23706818}.
CC -!- BIOTECHNOLOGY: Coexpression with both gRNAs or a synthetic gRNA in
CC human embryonic or pluripotent stem cells shows it is possible to
CC target and modify a DNA sequence of interest.
CC {ECO:0000269|PubMed:23940360}.
CC -!- SIMILARITY: Belongs to the CRISPR-associated protein Cas9 family.
CC Subtype II-C subfamily. {ECO:0000305}.
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DR EMBL; FM999788; CAX50779.1; -; Genomic_DNA.
DR RefSeq; WP_014574210.1; NC_017501.1.
DR PDB; 6J9M; X-ray; 2.39 A; A/F=51-123.
DR PDB; 6JDJ; X-ray; 2.60 A; C=1-77.
DR PDB; 6JDQ; X-ray; 2.95 A; A=1-1082.
DR PDB; 6JDV; X-ray; 3.10 A; A=1-1082.
DR PDB; 6JDX; X-ray; 2.28 A; C=1-77.
DR PDB; 6JE4; X-ray; 3.07 A; A/E/K/O=1-1082.
DR PDB; 6JE9; X-ray; 3.46 A; A/C=1-1082.
DR PDB; 6KC7; X-ray; 3.30 A; A=1-1082.
DR PDB; 6KC8; X-ray; 2.90 A; A=1-1082.
DR PDBsum; 6J9M; -.
DR PDBsum; 6JDJ; -.
DR PDBsum; 6JDQ; -.
DR PDBsum; 6JDV; -.
DR PDBsum; 6JDX; -.
DR PDBsum; 6JE4; -.
DR PDBsum; 6JE9; -.
DR PDBsum; 6KC7; -.
DR PDBsum; 6KC8; -.
DR AlphaFoldDB; C9X1G5; -.
DR SMR; C9X1G5; -.
DR KEGG; nmt:NMV_1993; -.
DR PATRIC; fig|604162.3.peg.2305; -.
DR HOGENOM; CLU_007514_0_0_4; -.
DR OMA; GQGHMET; -.
DR Proteomes; UP000002076; 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; IPR033114; HNH_CAS9.
DR InterPro; IPR003615; HNH_nuc.
DR InterPro; IPR036397; RNaseH_sf.
DR InterPro; IPR041383; RuvC_III.
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..1082
FT /note="CRISPR-associated endonuclease Cas9"
FT /id="PRO_0000429986"
FT DOMAIN 512..667
FT /note="HNH Cas9-type"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01085"
FT ACT_SITE 16
FT /note="For RuvC-like nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT ACT_SITE 588
FT /note="Proton acceptor for HNH nuclease domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 16
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 16
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 504
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 508
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 508
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT BINDING 723
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01480"
FT MUTAGEN 16
FT /note="D->A: Does not restore CRISPR interference during
FT plasmid transformation to deletion mutant."
FT /evidence="ECO:0000269|PubMed:23706818"
FT MUTAGEN 588
FT /note="H->A: Does not restore CRISPR interference during
FT plasmid transformation to deletion mutant."
FT /evidence="ECO:0000269|PubMed:23706818"
FT STRAND 11..17
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 18..21
FT /evidence="ECO:0007829|PDB:6JDJ"
FT STRAND 22..26
FT /evidence="ECO:0007829|PDB:6JDJ"
FT STRAND 31..33
FT /evidence="ECO:0007829|PDB:6JE4"
FT HELIX 38..41
FT /evidence="ECO:0007829|PDB:6JDX"
FT STRAND 47..50
FT /evidence="ECO:0007829|PDB:6JDX"
FT TURN 52..54
FT /evidence="ECO:0007829|PDB:6JDQ"
FT HELIX 56..73
FT /evidence="ECO:0007829|PDB:6JDX"
FT TURN 89..92
FT /evidence="ECO:0007829|PDB:6JE9"
FT STRAND 94..97
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 99..101
FT /evidence="ECO:0007829|PDB:6JE4"
FT STRAND 103..105
FT /evidence="ECO:0007829|PDB:6KC7"
FT HELIX 110..116
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 117..119
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 124..136
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 153..171
FT /evidence="ECO:0007829|PDB:6JDQ"
FT HELIX 177..188
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 196..198
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 205..221
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 230..240
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 248..250
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 251..254
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 259..261
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 264..266
FT /evidence="ECO:0007829|PDB:6JE9"
FT STRAND 268..271
FT /evidence="ECO:0007829|PDB:6JDQ"
FT HELIX 272..285
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 287..289
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 294..296
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 299..305
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 308..311
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 317..323
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 330..332
FT /evidence="ECO:0007829|PDB:6JDQ"
FT STRAND 338..340
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 342..344
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 345..348
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 351..362
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 368..370
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 376..388
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 392..399
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 400..402
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 405..412
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 420..424
FT /evidence="ECO:0007829|PDB:6JE4"
FT HELIX 425..436
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 441..449
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 450..453
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 461..463
FT /evidence="ECO:0007829|PDB:6JDQ"
FT STRAND 469..471
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 475..495
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 499..506
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 507..510
FT /evidence="ECO:0007829|PDB:6JDQ"
FT HELIX 513..540
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 541..543
FT /evidence="ECO:0007829|PDB:6JDQ"
FT STRAND 545..547
FT /evidence="ECO:0007829|PDB:6JE9"
FT HELIX 551..559
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 560..564
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 567..569
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 575..577
FT /evidence="ECO:0007829|PDB:6JDQ"
FT STRAND 578..580
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 583..590
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 592..594
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 602..607
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 609..612
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 619..622
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 623..626
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 630..640
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 646..652
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 667..684
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 690..692
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 695..699
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 700..709
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 716..719
FT /evidence="ECO:0007829|PDB:6JDQ"
FT HELIX 722..732
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 735..747
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 752..754
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 759..762
FT /evidence="ECO:0007829|PDB:6JDQ"
FT STRAND 767..769
FT /evidence="ECO:0007829|PDB:6JE9"
FT STRAND 775..778
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 779..788
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 803..813
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 814..816
FT /evidence="ECO:0007829|PDB:6JDQ"
FT STRAND 820..822
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 843..849
FT /evidence="ECO:0007829|PDB:6JDQ"
FT HELIX 853..855
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 857..863
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 864..866
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 869..872
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 876..878
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 882..894
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 895..897
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 899..902
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 907..910
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 912..914
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 916..928
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 933..935
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 936..939
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 940..942
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 947..955
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 958..965
FT /evidence="ECO:0007829|PDB:6KC8"
FT HELIX 966..970
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 981..983
FT /evidence="ECO:0007829|PDB:6JDQ"
FT HELIX 985..987
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 988..990
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 995..1001
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 1006..1010
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 1015..1023
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 1025..1027
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 1030..1033
FT /evidence="ECO:0007829|PDB:6KC8"
FT TURN 1039..1042
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 1046..1050
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 1056..1062
FT /evidence="ECO:0007829|PDB:6KC8"
FT STRAND 1065..1067
FT /evidence="ECO:0007829|PDB:6JDV"
FT STRAND 1071..1073
FT /evidence="ECO:0007829|PDB:6JE4"
SQ SEQUENCE 1082 AA; 124395 MW; 7250FDEE0C747F17 CRC64;
MAAFKPNSIN YILGLDIGIA SVGWAMVEID EEENPIRLID LGVRVFERAE VPKTGDSLAM
ARRLARSVRR LTRRRAHRLL RTRRLLKREG VLQAANFDEN GLIKSLPNTP WQLRAAALDR
KLTPLEWSAV LLHLIKHRGY LSQRKNEGET ADKELGALLK GVAGNAHALQ TGDFRTPAEL
ALNKFEKESG HIRNQRSDYS HTFSRKDLQA ELILLFEKQK EFGNPHVSGG LKEGIETLLM
TQRPALSGDA VQKMLGHCTF EPAEPKAAKN TYTAERFIWL TKLNNLRILE QGSERPLTDT
ERATLMDEPY RKSKLTYAQA RKLLGLEDTA FFKGLRYGKD NAEASTLMEM KAYHAISRAL
EKEGLKDKKS PLNLSPELQD EIGTAFSLFK TDEDITGRLK DRIQPEILEA LLKHISFDKF
VQISLKALRR IVPLMEQGKR YDEACAEIYG DHYGKKNTEE KIYLPPIPAD EIRNPVVLRA
LSQARKVING VVRRYGSPAR IHIETAREVG KSFKDRKEIE KRQEENRKDR EKAAAKFREY
FPNFVGEPKS KDILKLRLYE QQHGKCLYSG KEINLGRLNE KGYVEIDHAL PFSRTWDDSF
NNKVLVLGSE NQNKGNQTPY EYFNGKDNSR EWQEFKARVE TSRFPRSKKQ RILLQKFDED
GFKERNLNDT RYVNRFLCQF VADRMRLTGK GKKRVFASNG QITNLLRGFW GLRKVRAEND
RHHALDAVVV ACSTVAMQQK ITRFVRYKEM NAFDGKTIDK ETGEVLHQKT HFPQPWEFFA
QEVMIRVFGK PDGKPEFEEA DTLEKLRTLL AEKLSSRPEA VHEYVTPLFV SRAPNRKMSG
QGHMETVKSA KRLDEGVSVL RVPLTQLKLK DLEKMVNRER EPKLYEALKA RLEAHKDDPA
KAFAEPFYKY DKAGNRTQQV KAVRVEQVQK TGVWVRNHNG IADNATMVRV DVFEKGDKYY
LVPIYSWQVA KGILPDRAVV QGKDEEDWQL IDDSFNFKFS LHPNDLVEVI TKKARMFGYF
ASCHRGTGNI NIRIHDLDHK IGKNGILEGI GVKTALSFQK YQIDELGKEI RPCRLKKRPP
VR
