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Q99ZW2 (CAS9_STRP1) Reviewed, UniProtKB/Swiss-Prot

Last modified May 14, 2014. Version 72. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
CRISPR-associated endonuclease Cas9/Csn1

EC=3.1.-.-
Gene names
Name:cas9
Synonyms:csn1
Ordered Locus Names:SPy_1046
OrganismStreptococcus pyogenes serotype M1 [Complete proteome] [HAMAP]
Taxonomic identifier301447 [NCBI]
Taxonomic lineageBacteriaFirmicutesBacilliLactobacillalesStreptococcaceaeStreptococcus

Protein attributes

Sequence length1368 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA) Probable. In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer. The target strand not complementary to crRNA is first cut endonucleolytically, then trimmed by 3'-5' exonucleolytically. DNA-binding requires protein and both RNA species. Cas9 probably recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself. Ref.2 Ref.3

Cofactor

Endonuclease activity on target dsDNA requires Mg2+. Ref.3

Subunit structure

Monomer By similarity. Binds crRNA and tracrRNA. HAMAP-Rule MF_01480

Domain

Has 2 endonuclease domains. The RuvC-like domain (residues 7-22, 759-766 and 982-989) cleaves the target DNA noncomplementary to crRNA while the HNH nuclease domain (residues 810-872) cleaves the target DNA complementary to crRNA. HAMAP-Rule MF_01480

Disruption phenotype

Loss of correct processing of pre-crRNA and tracrRNA. Loss of immunity against a plasmid with homology to CRISPR spacer sequences. Ref.2

Biotechnological use

Coexpression of Cas9 with an artifical guide RNA (gRNAs) which fuses the crRNA with the tracrRNA in human cells has shown it is possible to target and modify a DNA sequence of interest (Ref.5, Ref.8, Ref.9). Cas9 plus the 2 RNA species have also been expressed individually in human and mouse cells to achieve DNA targeting; cleavage efficiencies of the artificial construct were lower that those for systems with the crRNA and tracrRNA expressed separately (Ref.4). Microinjection of Cas9-encoding mRNA and a synthetic guide RNA fusing crRNA and tracrRNA into zebrafish embryos induces targeted mutations (Ref.6). In all cases introduction of multiple gRNAs leads to multiplexed editing of genomic loci; DNA has also been inserted into a mammalian locus of interest. In S.pneumoniae and E.coli it has been used to generate markerless mutations; mutiple changes can be made simultaneously (Ref.7). Ref.3 Ref.4 Ref.5 Ref.6 Ref.7 Ref.8 Ref.9

Sequence similarities

Belongs to the CRISPR-associated Cas9/Csn1 family. Subtype II/Nmemi subfamily.

Contains 1 HNH Cas9-type domain.

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 13681368CRISPR-associated endonuclease Cas9/Csn1 HAMAP-Rule MF_01480
PRO_0000418437

Regions

Domain810 – 87263HNH Cas9-type

Experimental info

Mutagenesis101D → A: Target DNA noncomplementary to the crRNA is not cleaved. Ref.3
Mutagenesis8401H → A: Target DNA complementary to the crRNA is not cleaved. Ref.3
Mutagenesis11321G → C: Probably inactivates protein. Ref.7

Secondary structure

.................................................................................................................................................................................................................................................. 1368
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
Q99ZW2 [UniParc].

Last modified June 1, 2001. Version 1.
Checksum: 07D04F0B5965762F

FASTA1,368158,441
        10         20         30         40         50         60 
MDKKYSIGLD IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE 

        70         80         90        100        110        120 
ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG 

       130        140        150        160        170        180 
NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD 

       190        200        210        220        230        240 
VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN 

       250        260        270        280        290        300 
LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI 

       310        320        330        340        350        360 
LLSDILRVNT EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA 

       370        380        390        400        410        420 
GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH 

       430        440        450        460        470        480 
AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE 

       490        500        510        520        530        540 
VVDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL 

       550        560        570        580        590        600 
SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI 

       610        620        630        640        650        660 
IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG 

       670        680        690        700        710        720 
RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL 

       730        740        750        760        770        780 
HEHIANLAGS PAIKKGILQT VKVVDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER 

       790        800        810        820        830        840 
MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDH 

       850        860        870        880        890        900 
IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEVVKKMK NYWRQLLNAK LITQRKFDNL 

       910        920        930        940        950        960 
TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS 

       970        980        990       1000       1010       1020 
KLVSDFRKDF QFYKVREINN YHHAHDAYLN AVVGTALIKK YPKLESEFVY GDYKVYDVRK 

      1030       1040       1050       1060       1070       1080 
MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF 

      1090       1100       1110       1120       1130       1140 
ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA 

      1150       1160       1170       1180       1190       1200 
YSVLVVAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK 

      1210       1220       1230       1240       1250       1260 
YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE 

      1270       1280       1290       1300       1310       1320 
QHKHYLDEII EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA 

      1330       1340       1350       1360 
PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD 

« Hide

References

« Hide 'large scale' references
[1]"Complete genome sequence of an M1 strain of Streptococcus pyogenes."
Ferretti J.J., McShan W.M., Ajdic D.J., Savic D.J., Savic G., Lyon K., Primeaux C., Sezate S., Suvorov A.N., Kenton S., Lai H.S., Lin S.P., Qian Y., Jia H.G., Najar F.Z., Ren Q., Zhu H., Song L. expand/collapse author list , White J., Yuan X., Clifton S.W., Roe B.A., McLaughlin R.E.
Proc. Natl. Acad. Sci. U.S.A. 98:4658-4663(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: ATCC 700294 / SF370 / Serotype M1.
[2]"CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III."
Deltcheva E., Chylinski K., Sharma C.M., Gonzales K., Chao Y., Pirzada Z.A., Eckert M.R., Vogel J., Charpentier E.
Nature 471:602-607(2011) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION IN CRISPR-MEDIATED PLASMID DEFENSE, DISRUPTION PHENOTYPE.
Strain: ATCC 700294 / SF370 / Serotype M1.
[3]"A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity."
Jinek M., Chylinski K., Fonfara I., Hauer M., Doudna J.A., Charpentier E.
Science 337:816-821(2012) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION AS A DNA ENDONUCLEASE, FUNCTION AS AN EXONUCLEASE, COFACTOR, DNA-BINDING, POSSIBLE BIOTECHNOLOGY, MUTAGENESIS OF ASP-10 AND HIS-840.
Strain: ATCC 700294 / SF370 / Serotype M1.
[4]"Multiplex genome engineering using CRISPR/Cas systems."
Cong L., Ran F.A., Cox D., Lin S., Barretto R., Habib N., Hsu P.D., Wu X., Jiang W., Marraffini L.A., Zhang F.
Science 339:819-823(2013) [PubMed] [Europe PMC] [Abstract]
Cited for: BIOTECHNOLOGY IN HUMAN AND MOUSE CELLS.
Strain: ATCC 700294 / SF370 / Serotype M1.
[5]"RNA-guided human genome engineering via Cas9."
Mali P., Yang L., Esvelt K.M., Aach J., Guell M., Dicarlo J.E., Norville J.E., Church G.M.
Science 339:823-826(2013) [PubMed] [Europe PMC] [Abstract]
Cited for: BIOTECHNOLOGY IN HUMAN CELLS.
[6]"Efficient genome editing in zebrafish using a CRISPR-Cas system."
Hwang W.Y., Fu Y., Reyon D., Maeder M.L., Tsai S.Q., Sander J.D., Peterson R.T., Yeh J.R., Joung J.K.
Nat. Biotechnol. 31:227-229(2013) [PubMed] [Europe PMC] [Abstract]
Cited for: BIOTECHNOLOGY IN ZEBRAFISH EMBRYOS.
[7]"RNA-guided editing of bacterial genomes using CRISPR-Cas systems."
Jiang W., Bikard D., Cox D., Zhang F., Marraffini L.A.
Nat. Biotechnol. 31:233-239(2013) [PubMed] [Europe PMC] [Abstract]
Cited for: BIOTECHNOLOGY IN BACTERIA, MUTAGENESIS OF GLY-1132.
Strain: ATCC 700294 / SF370 / Serotype M1.
[8]"Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease."
Cho S.W., Kim S., Kim J.M., Kim J.S.
Nat. Biotechnol. 31:230-232(2013) [PubMed] [Europe PMC] [Abstract]
Cited for: BIOTECHNOLOGY IN HUMAN CELLS.
Strain: ATCC 700294 / SF370 / Serotype M1.
[9]"RNA-programmed genome editing in human cells."
Jinek M., East A., Cheng A., Lin S., Ma E., Doudna J.
Elife 2:E00471-E00471(2013) [PubMed] [Europe PMC] [Abstract]
Cited for: BIOTECHNOLOGY IN HUMAN CELLS.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
AE004092 Genomic DNA. Translation: AAK33936.1.
RefSeqNP_269215.1. NC_002737.1.

3D structure databases

PDBe
RCSB-PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
4CMPX-ray2.62A/B1-1368[»]
4CMQX-ray3.09A/B1-1368[»]
4OO8X-ray2.50A/D1-1368[»]
ProteinModelPortalQ99ZW2.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

STRING160490.SPy_1046.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaAAK33936; AAK33936; SPy_1046.
GeneID901176.
KEGGspy:SPy_1046.
PATRIC19715675. VBIStrPyo79812_0902.

Organism-specific databases

CMRSearch...

Phylogenomic databases

HOGENOMHOG000071789.
KOK09952.
OMAIPHQIHL.
OrthoDBEOG6MH57S.

Enzyme and pathway databases

BioCycSPYO160490:GJ81-863-MONOMER.

Family and domain databases

HAMAPMF_01480. Cas9.
InterProIPR028629. Cas9/Csn1_fam.
IPR025978. CRISPR-rel_HNH_dom.
IPR003615. HNH_nuc.
[Graphical view]
PfamPF13395. HNH_4. 1 hit.
[Graphical view]
SMARTSM00507. HNHc. 1 hit.
[Graphical view]
TIGRFAMsTIGR01865. cas_Csn1. 1 hit.
ProtoNetSearch...

Entry information

Entry nameCAS9_STRP1
AccessionPrimary (citable) accession number: Q99ZW2
Entry history
Integrated into UniProtKB/Swiss-Prot: July 11, 2012
Last sequence update: June 1, 2001
Last modified: May 14, 2014
This is version 72 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references