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Protein

CRISPR-associated endonuclease Cas9 2

Gene

cas9-2

Organism
Streptococcus thermophilus (strain ATCC BAA-491 / LMD-9)
Status
Reviewed-Annotation score: Annotation score: 4 out of 5-Experimental evidence at protein leveli

Functioni

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). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein (By similarity). 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; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). Cas9 recognizes the protospacer adjacent motif (PAM) in the CRISPR repeat sequences to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. PAM recognition is also required for catalytic activity. Complements the gRNA coprocessing defect in a cas9 deletion in S.pyogenes strain 370, and cuts target DNA in Cas9:gRNAs mixing experiments with S.mutans strain UA159.UniRule annotation3 Publications

Cofactori

Mg2+CuratedNote: Endonuclease activity on target dsDNA requires Mg2+.Curated

Sites

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Active sitei10For RuvC-like nuclease domainUniRule annotation1
Metal bindingi10Magnesium 1UniRule annotation1
Metal bindingi10Magnesium 2UniRule annotation1
Metal bindingi763Magnesium 1UniRule annotation1
Metal bindingi767Magnesium 1UniRule annotation1
Metal bindingi767Magnesium 2UniRule annotation1
Active sitei847Proton acceptor for HNH nuclease domainUniRule annotation1
Metal bindingi990Magnesium 2; via pros nitrogenUniRule annotation1

GO - Molecular functioni

GO - Biological processi

Keywordsi

Molecular functionDNA-binding, Endonuclease, Hydrolase, Nuclease, RNA-binding
Biological processAntiviral defense
LigandMagnesium, Manganese, Metal-binding

Names & Taxonomyi

Protein namesi
Recommended name:
CRISPR-associated endonuclease Cas9 2UniRule annotation (EC:3.1.-.-UniRule annotation)
Alternative name(s):
Cas9*
St3Cas9
Gene namesi
Name:cas9-2UniRule annotation
Synonyms:csn1
Ordered Locus Names:STER_1477
OrganismiStreptococcus thermophilus (strain ATCC BAA-491 / LMD-9)
Taxonomic identifieri322159 [NCBI]
Taxonomic lineageiBacteriaFirmicutesBacilliLactobacillalesStreptococcaceaeStreptococcus

Pathology & Biotechi

Biotechnological usei

Coexpression of Cas9 and both gRNAs in human cells has shown it is possible to use this system to target and modify a DNA sequence of interest in situ.1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)DescriptionActionsGraphical viewLength
ChainiPRO_00004216861 – 1388CRISPR-associated endonuclease Cas9 2Add BLAST1388

Interactioni

Subunit structurei

Monomer (By similarity). Binds crRNA and tracrRNA.UniRule annotation1 Publication

Structurei

3D structure databases

ProteinModelPortaliQ03JI6.
SMRiQ03JI6.
ModBaseiSearch...
MobiDBiSearch...

Family & Domainsi

Domains and Repeats

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Domaini771 – 928HNH Cas9-typePROSITE-ProRule annotationAdd BLAST158

Region

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Regioni1102 – 1388PAM-interacting domain (PI)Add BLAST287

Domaini

Has 2 endonuclease domains. The discontinuous RuvC-like domain cleaves the target DNA noncomplementary to crRNA while the HNH nuclease domain cleaves the target DNA complementary to crRNA.UniRule annotation
The PAM-interacting domain (PI domain, approximately residues 1102-1388) recognizes the PAM motif; swapping the PI domain of this enzyme with that from S.pyogenes Cas9 (AC Q99ZW2) prevents cleavage of DNA with the endogenous PAM site but confers the ability to cleave DNA with the PAM site specific for S.pyogenes CRISPRs.1 Publication

Sequence similaritiesi

Belongs to the CRISPR-associated protein Cas9 family. Subtype II-A subfamily.UniRule annotationCurated

Phylogenomic databases

HOGENOMiHOG000071789.
KOiK09952.
OMAiTDRHSIK.

Family and domain databases

HAMAPiMF_01480. Cas9. 1 hit.
InterProiView protein in InterPro
IPR028629. Cas9.
IPR032239. Cas9-BH.
IPR032237. Cas9_PI.
IPR032240. Cas9_REC.
IPR033114. HNH_CAS9.
IPR003615. HNH_nuc.
PfamiView protein in Pfam
PF16593. Cas9-BH. 1 hit.
PF16595. Cas9_PI. 1 hit.
PF16592. Cas9_REC. 1 hit.
PF13395. HNH_4. 1 hit.
TIGRFAMsiTIGR01865. cas_Csn1. 1 hit.
PROSITEiView protein in PROSITE
PS51749. HNH_CAS9. 1 hit.

Sequencei

Sequence statusi: Complete.

Q03JI6-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MTKPYSIGLD IGTNSVGWAV TTDNYKVPSK KMKVLGNTSK KYIKKNLLGV
60 70 80 90 100
LLFDSGITAE GRRLKRTARR RYTRRRNRIL YLQEIFSTEM ATLDDAFFQR
110 120 130 140 150
LDDSFLVPDD KRDSKYPIFG NLVEEKAYHD EFPTIYHLRK YLADSTKKAD
160 170 180 190 200
LRLVYLALAH MIKYRGHFLI EGEFNSKNND IQKNFQDFLD TYNAIFESDL
210 220 230 240 250
SLENSKQLEE IVKDKISKLE KKDRILKLFP GEKNSGIFSE FLKLIVGNQA
260 270 280 290 300
DFRKCFNLDE KASLHFSKES YDEDLETLLG YIGDDYSDVF LKAKKLYDAI
310 320 330 340 350
LLSGFLTVTD NETEAPLSSA MIKRYNEHKE DLALLKEYIR NISLKTYNEV
360 370 380 390 400
FKDDTKNGYA GYIDGKTNQE DFYVYLKKLL AEFEGADYFL EKIDREDFLR
410 420 430 440 450
KQRTFDNGSI PYQIHLQEMR AILDKQAKFY PFLAKNKERI EKILTFRIPY
460 470 480 490 500
YVGPLARGNS DFAWSIRKRN EKITPWNFED VIDKESSAEA FINRMTSFDL
510 520 530 540 550
YLPEEKVLPK HSLLYETFNV YNELTKVRFI AESMRDYQFL DSKQKKDIVR
560 570 580 590 600
LYFKDKRKVT DKDIIEYLHA IYGYDGIELK GIEKQFNSSL STYHDLLNII
610 620 630 640 650
NDKEFLDDSS NEAIIEEIIH TLTIFEDREM IKQRLSKFEN IFDKSVLKKL
660 670 680 690 700
SRRHYTGWGK LSAKLINGIR DEKSGNTILD YLIDDGISNR NFMQLIHDDA
710 720 730 740 750
LSFKKKIQKA QIIGDEDKGN IKEVVKSLPG SPAIKKGILQ SIKIVDELVK
760 770 780 790 800
VMGGRKPESI VVEMARENQY TNQGKSNSQQ RLKRLEKSLK ELGSKILKEN
810 820 830 840 850
IPAKLSKIDN NALQNDRLYL YYLQNGKDMY TGDDLDIDRL SNYDIDHIIP
860 870 880 890 900
QAFLKDNSID NKVLVSSASN RGKSDDVPSL EVVKKRKTFW YQLLKSKLIS
910 920 930 940 950
QRKFDNLTKA ERGGLSPEDK AGFIQRQLVE TRQITKHVAR LLDEKFNNKK
960 970 980 990 1000
DENNRAVRTV KIITLKSTLV SQFRKDFELY KVREINDFHH AHDAYLNAVV
1010 1020 1030 1040 1050
ASALLKKYPK LEPEFVYGDY PKYNSFRERK SATEKVYFYS NIMNIFKKSI
1060 1070 1080 1090 1100
SLADGRVIER PLIEVNEETG ESVWNKESDL ATVRRVLSYP QVNVVKKVEE
1110 1120 1130 1140 1150
QNHGLDRGKP KGLFNANLSS KPKPNSNENL VGAKEYLDPK KYGGYAGISN
1160 1170 1180 1190 1200
SFTVLVKGTI EKGAKKKITN VLEFQGISIL DRINYRKDKL NFLLEKGYKD
1210 1220 1230 1240 1250
IELIIELPKY SLFELSDGSR RMLASILSTN NKRGEIHKGN QIFLSQKFVK
1260 1270 1280 1290 1300
LLYHAKRISN TINENHRKYV ENHKKEFEEL FYYILEFNEN YVGAKKNGKL
1310 1320 1330 1340 1350
LNSAFQSWQN HSIDELCSSF IGPTGSERKG LFELTSRGSA ADFEFLGVKI
1360 1370 1380
PRYRDYTPSS LLKDATLIHQ SVTGLYETRI DLAKLGEG
Length:1,388
Mass (Da):161,031
Last modified:November 14, 2006 - v1
Checksum:iB97BC4C953090235
GO

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
CP000419 Genomic DNA. Translation: ABJ66636.1.
RefSeqiWP_011681470.1. NC_008532.1.

Genome annotation databases

EnsemblBacteriaiABJ66636; ABJ66636; STER_1477.
KEGGiste:STER_1477.

Similar proteinsi

Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.

Entry informationi

Entry nameiCAS9B_STRTD
AccessioniPrimary (citable) accession number: Q03JI6
Entry historyiIntegrated into UniProtKB/Swiss-Prot: March 6, 2013
Last sequence update: November 14, 2006
Last modified: June 7, 2017
This is version 66 of the entry and version 1 of the sequence. See complete history.
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Documents

  1. SIMILARITY comments
    Index of protein domains and families