UniProtKB - G3ECR1 (CAS9_STRTR)
Protein
CRISPR-associated endonuclease Cas9
Gene
cas9
Organism
Streptococcus thermophilus
Status
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 Cas9. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA (Probable). Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer yielding blunt ends; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). Cas9 recognizes a 3'-G-rich protospacer adjacent motif (PAM, TGGTG in this organism) 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. When the CRISPR3/cas system consisting of cas9-cas1-cas2-csn2-CRISPR3 or just cas9-CRISPR3 is expressed in E.coli it prevents plasmids homologous to spacers 1 or 2 from transforming.Curated3 Publications
Miscellaneous
This strain encodes 4 CRISPR-Cas systems; this is CRISPR3.
Caution
It is uncertain whether Met-1 or Met-22 is the initiator.Curated
Cofactori
Mg2+1 PublicationNote: Endonuclease activity on target DNA requires Mg2+.1 Publication
Activity regulationi
Only has nuclease activity when bound to both gRNAs (crRNA plus tracrRNA).1 Publication
Sites
Feature key | Position(s) | DescriptionActions | Graphical view | Length |
---|---|---|---|---|
Active sitei | 31 | For RuvC-like nuclease domainUniRule annotation | 1 | |
Metal bindingi | 31 | Magnesium 1UniRule annotation | 1 | |
Metal bindingi | 31 | Magnesium 2UniRule annotation | 1 | |
Metal bindingi | 784 | Magnesium 1UniRule annotation | 1 | |
Metal bindingi | 788 | Magnesium 1UniRule annotation | 1 | |
Metal bindingi | 788 | Magnesium 2UniRule annotation | 1 | |
Active sitei | 868 | Proton acceptor for HNH nuclease domainUniRule annotation | 1 | |
Metal bindingi | 1011 | Magnesium 2; via pros nitrogenUniRule annotation | 1 |
GO - Molecular functioni
- DNA binding Source: UniProtKB-KW
- endonuclease activity Source: UniProtKB-UniRule
- metal ion binding Source: UniProtKB-UniRule
- RNA binding Source: UniProtKB-KW
GO - Biological processi
- defense response to virus Source: UniProtKB-UniRule
- maintenance of CRISPR repeat elements Source: UniProtKB-UniRule
Keywordsi
Molecular function | DNA-binding, Endonuclease, Hydrolase, Nuclease, RNA-binding |
Biological process | Antiviral defense |
Ligand | Magnesium, Manganese, Metal-binding |
Names & Taxonomyi
Protein namesi | Recommended name: CRISPR-associated endonuclease Cas9UniRule annotation (EC:3.1.-.-UniRule annotation)Alternative name(s): St-Cas9 |
Gene namesi | Name:cas9UniRule annotation Synonyms:csn1 |
Organismi | Streptococcus thermophilus |
Taxonomic identifieri | 1308 [NCBI] |
Taxonomic lineagei | Bacteria › Firmicutes › Bacilli › Lactobacillales › Streptococcaceae › Streptococcus |
Pathology & Biotechi
Biotechnological usei
The simplicity of the Cas9-gRNAs RNA-directed DNA endonuclease activity may be used to target and modify a DNA sequence of interest.
Disruption phenotypei
Plasmid transformation is restored.1 Publication
Mutagenesis
Feature key | Position(s) | DescriptionActions | Graphical view | Length |
---|---|---|---|---|
Mutagenesisi | 31 | D → A: No longer prevents plasmid transformation. Target DNA noncomplementary to the crRNA is not cleaved. 2 Publications | 1 | |
Mutagenesisi | 868 | H → A: No longer prevents plasmid transformation. Target DNA complementary to the crRNA is not cleaved. 1 Publication | 1 | |
Mutagenesisi | 882 | N → A: No longer prevents plasmid transformation. 1 Publication | 1 | |
Mutagenesisi | 891 | N → A: No longer prevents plasmid transformation. 2 Publications | 1 |
PTM / Processingi
Molecule processing
Feature key | Position(s) | DescriptionActions | Graphical view | Length |
---|---|---|---|---|
ChainiPRO_0000417879 | 1 – 1409 | CRISPR-associated endonuclease Cas9Add BLAST | 1409 |
Proteomic databases
PRIDEi | G3ECR1 |
Interactioni
Subunit structurei
Monomer (Probable). Binds crRNA and tracrRNA.
UniRule annotationCurated1 PublicationProtein-protein interaction databases
STRINGi | 322159.STER_1477 |
Family & Domainsi
Domains and Repeats
Feature key | Position(s) | DescriptionActions | Graphical view | Length |
---|---|---|---|---|
Domaini | 792 – 949 | HNH Cas9-typePROSITE-ProRule annotationAdd BLAST | 158 |
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
Sequence similaritiesi
Phylogenomic databases
eggNOGi | COG3513, Bacteria |
Family and domain databases
HAMAPi | MF_01480, Cas9, 1 hit |
InterProi | View protein in InterPro IPR028629, Cas9 IPR032239, Cas9-BH IPR032237, Cas9_PI IPR032240, Cas9_REC IPR033114, HNH_CAS9 IPR003615, HNH_nuc |
Pfami | View protein in Pfam PF16593, Cas9-BH, 1 hit PF16595, Cas9_PI, 1 hit PF16592, Cas9_REC, 1 hit PF13395, HNH_4, 1 hit |
TIGRFAMsi | TIGR01865, cas_Csn1, 1 hit |
PROSITEi | View protein in PROSITE PS51749, HNH_CAS9, 1 hit |
i Sequence
Sequence statusi: Complete.
G3ECR1-1 [UniParc]FASTAAdd to basket
10 20 30 40 50
MLFNKCIIIS INLDFSNKEK CMTKPYSIGL DIGTNSVGWA VITDNYKVPS
60 70 80 90 100
KKMKVLGNTS KKYIKKNLLG VLLFDSGITA EGRRLKRTAR RRYTRRRNRI
110 120 130 140 150
LYLQEIFSTE MATLDDAFFQ RLDDSFLVPD DKRDSKYPIF GNLVEEKVYH
160 170 180 190 200
DEFPTIYHLR KYLADSTKKA DLRLVYLALA HMIKYRGHFL IEGEFNSKNN
210 220 230 240 250
DIQKNFQDFL DTYNAIFESD LSLENSKQLE EIVKDKISKL EKKDRILKLF
260 270 280 290 300
PGEKNSGIFS EFLKLIVGNQ ADFRKCFNLD EKASLHFSKE SYDEDLETLL
310 320 330 340 350
GYIGDDYSDV FLKAKKLYDA ILLSGFLTVT DNETEAPLSS AMIKRYNEHK
360 370 380 390 400
EDLALLKEYI RNISLKTYNE VFKDDTKNGY AGYIDGKTNQ EDFYVYLKNL
410 420 430 440 450
LAEFEGADYF LEKIDREDFL RKQRTFDNGS IPYQIHLQEM RAILDKQAKF
460 470 480 490 500
YPFLAKNKER IEKILTFRIP YYVGPLARGN SDFAWSIRKR NEKITPWNFE
510 520 530 540 550
DVIDKESSAE AFINRMTSFD LYLPEEKVLP KHSLLYETFN VYNELTKVRF
560 570 580 590 600
IAESMRDYQF LDSKQKKDIV RLYFKDKRKV TDKDIIEYLH AIYGYDGIEL
610 620 630 640 650
KGIEKQFNSS LSTYHDLLNI INDKEFLDDS SNEAIIEEII HTLTIFEDRE
660 670 680 690 700
MIKQRLSKFE NIFDKSVLKK LSRRHYTGWG KLSAKLINGI RDEKSGNTIL
710 720 730 740 750
DYLIDDGISN RNFMQLIHDD ALSFKKKIQK AQIIGDEDKG NIKEVVKSLP
760 770 780 790 800
GSPAIKKGIL QSIKIVDELV KVMGGRKPES IVVEMARENQ YTNQGKSNSQ
810 820 830 840 850
QRLKRLEKSL KELGSKILKE NIPAKLSKID NNALQNDRLY LYYLQNGKDM
860 870 880 890 900
YTGDDLDIDR LSNYDIDHII PQAFLKDNSI DNKVLVSSAS NRGKSDDFPS
910 920 930 940 950
LEVVKKRKTF WYQLLKSKLI SQRKFDNLTK AERGGLLPED KAGFIQRQLV
960 970 980 990 1000
ETRQITKHVA RLLDEKFNNK KDENNRAVRT VKIITLKSTL VSQFRKDFEL
1010 1020 1030 1040 1050
YKVREINDFH HAHDAYLNAV IASALLKKYP KLEPEFVYGD YPKYNSFRER
1060 1070 1080 1090 1100
KSATEKVYFY SNIMNIFKKS ISLADGRVIE RPLIEVNEET GESVWNKESD
1110 1120 1130 1140 1150
LATVRRVLSY PQVNVVKKVE EQNHGLDRGK PKGLFNANLS SKPKPNSNEN
1160 1170 1180 1190 1200
LVGAKEYLDP KKYGGYAGIS NSFAVLVKGT IEKGAKKKIT NVLEFQGISI
1210 1220 1230 1240 1250
LDRINYRKDK LNFLLEKGYK DIELIIELPK YSLFELSDGS RRMLASILST
1260 1270 1280 1290 1300
NNKRGEIHKG NQIFLSQKFV KLLYHAKRIS NTINENHRKY VENHKKEFEE
1310 1320 1330 1340 1350
LFYYILEFNE NYVGAKKNGK LLNSAFQSWQ NHSIDELCSS FIGPTGSERK
1360 1370 1380 1390 1400
GLFELTSRGS AADFEFLGVK IPRYRDYTPS SLLKDATLIH QSVTGLYETR
IDLAKLGEG
Sequence cautioni
The sequence AEM62887 differs from that shown. Reason: Erroneous initiation. Truncated N-terminus.Curated
Sequence databases
Select the link destinations: EMBLi GenBanki DDBJi Links Updated | HQ712120 Genomic DNA Translation: AEM62887.1 Different initiation. |
RefSeqi | WP_024703962.1, NZ_WMLD01000001.1 |
Similar proteinsi
Cross-referencesi
Sequence databases
Select the link destinations: EMBLi GenBanki DDBJi Links Updated | HQ712120 Genomic DNA Translation: AEM62887.1 Different initiation. |
RefSeqi | WP_024703962.1, NZ_WMLD01000001.1 |
3D structure databases
SMRi | G3ECR1 |
ModBasei | Search... |
Protein-protein interaction databases
STRINGi | 322159.STER_1477 |
Proteomic databases
PRIDEi | G3ECR1 |
Phylogenomic databases
eggNOGi | COG3513, Bacteria |
Family and domain databases
HAMAPi | MF_01480, Cas9, 1 hit |
InterProi | View protein in InterPro IPR028629, Cas9 IPR032239, Cas9-BH IPR032237, Cas9_PI IPR032240, Cas9_REC IPR033114, HNH_CAS9 IPR003615, HNH_nuc |
Pfami | View protein in Pfam PF16593, Cas9-BH, 1 hit PF16595, Cas9_PI, 1 hit PF16592, Cas9_REC, 1 hit PF13395, HNH_4, 1 hit |
TIGRFAMsi | TIGR01865, cas_Csn1, 1 hit |
PROSITEi | View protein in PROSITE PS51749, HNH_CAS9, 1 hit |
ProtoNeti | Search... |
MobiDBi | Search... |
Entry informationi
Entry namei | CAS9_STRTR | |
Accessioni | G3ECR1Primary (citable) accession number: G3ECR1 | |
Entry historyi | Integrated into UniProtKB/Swiss-Prot: | June 13, 2012 |
Last sequence update: | June 13, 2012 | |
Last modified: | August 12, 2020 | |
This is version 35 of the entry and version 2 of the sequence. See complete history. | ||
Entry statusi | Reviewed (UniProtKB/Swiss-Prot) | |
Annotation program | Prokaryotic Protein Annotation Program |