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Database: UniProt
Entry: P38036
LinkDB: P38036
Original site: P38036 
ID   CAS3_ECOLI              Reviewed;         888 AA.
AC   P38036; Q2MA68; Q46902;
DT   01-OCT-1994, integrated into UniProtKB/Swiss-Prot.
DT   01-NOV-1997, sequence version 2.
DT   13-NOV-2019, entry version 138.
DE   RecName: Full=CRISPR-associated endonuclease/helicase Cas3;
DE            EC=3.1.-.-;
DE            EC=3.6.4.-;
GN   Name=ygcB; Synonyms=cas3; OrderedLocusNames=b2761, JW2731;
OS   Escherichia coli (strain K12).
OC   Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC   Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=83333;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA   Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA   Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA   Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J.,
RA   Mau B., Shao Y.;
RT   "The complete genome sequence of Escherichia coli K-12.";
RL   Science 277:1453-1462(1997).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=16738553; DOI=10.1038/msb4100049;
RA   Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA   Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT   "Highly accurate genome sequences of Escherichia coli K-12 strains
RT   MG1655 and W3110.";
RL   Mol. Syst. Biol. 2:E1-E5(2006).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-335.
RC   STRAIN=K12;
RX   PubMed=2005873; DOI=10.1007/bf00269864;
RA   Krone F.A., Westphal G., Schwenn J.D.;
RT   "Characterisation of the gene cysH and of its product phospho-
RT   adenylylsulphate reductase from Escherichia coli.";
RL   Mol. Gen. Genet. 225:314-319(1991).
RN   [4]
RP   IDENTIFICATION.
RX   PubMed=7984428; DOI=10.1093/nar/22.22.4756;
RA   Borodovsky M., Rudd K.E., Koonin E.V.;
RT   "Intrinsic and extrinsic approaches for detecting genes in a bacterial
RT   genome.";
RL   Nucleic Acids Res. 22:4756-4767(1994).
RN   [5]
RP   FUNCTION, AND DISRUPTION PHENOTYPE.
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=18703739; DOI=10.1126/science.1159689;
RA   Brouns S.J., Jore M.M., Lundgren M., Westra E.R., Slijkhuis R.J.,
RA   Snijders A.P., Dickman M.J., Makarova K.S., Koonin E.V.,
RA   van der Oost J.;
RT   "Small CRISPR RNAs guide antiviral defense in prokaryotes.";
RL   Science 321:960-964(2008).
RN   [6]
RP   FUNCTION AS A HELICASE, FUNCTION IN FORMING R-LOOPS, COFACTOR,
RP   DNA-BINDING, AND MUTAGENESIS OF HIS-74; ASP-75; LYS-78;
RP   317-GLY-SER-318; LYS-320 AND HIS-455.
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=21699496; DOI=10.1042/bj20110901;
RA   Howard J.A., Delmas S., Ivancic-Bace I., Bolt E.L.;
RT   "Helicase dissociation and annealing of RNA-DNA hybrids by Escherichia
RT   coli Cas3 protein.";
RL   Biochem. J. 439:85-95(2011).
RN   [7]
RP   FUNCTION AS AN ENDONUCLEASE, FUNCTION AS AN EXONUCLEASE, COFACTOR,
RP   INTERACTION WITH CASA, AND MUTAGENESIS OF HIS-74; ASP-75; LYS-78;
RP   LYS-320; ASP-452 AND 483-SER--THR-485.
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=22521689; DOI=10.1016/j.molcel.2012.03.018;
RA   Westra E.R., van Erp P.B., Kunne T., Wong S.P., Staals R.H.,
RA   Seegers C.L., Bollen S., Jore M.M., Semenova E., Severinov K.,
RA   de Vos W.M., Dame R.T., de Vries R., Brouns S.J., van der Oost J.;
RT   "CRISPR immunity relies on the consecutive binding and degradation of
RT   negatively supercoiled invader DNA by Cascade and Cas3.";
RL   Mol. Cell 46:595-605(2012).
CC   -!- FUNCTION: CRISPR (clustered regularly interspaced short
CC       palindromic repeat), is an adaptive immune system that provides
CC       protection against mobile genetic elements (viruses, transposable
CC       elements and conjugative plasmids). CRISPR clusters contain
CC       sequences complementary to antecedent mobile elements and target
CC       invading nucleic acids. CRISPR clusters are transcribed and
CC       processed into CRISPR RNA (crRNA). Cas3 plus Cascade participate
CC       in CRISPR interference, the third stage of CRISPR immunity.
CC   -!- FUNCTION: Acts as an endonuclease, a 3'-5'exonuclease, and an ATP-
CC       dependent dsDNA helicase. Anneals and unwinds R-loops (in which
CC       crRNA binds the target DNA, displacing the noncomplementary
CC       strand). Unwinding requires ATP, annealing does not. Required
CC       along with the Cascade complex for resistance to bacteriophage
CC       lambda infection as well as the ability to cure CRISPR-encoding
CC       high-copy number plasmid. A Cas3-CasA fusion protein purified with
CC       the Cascade complex nicks target plasmid in the presence but not
CC       absence of Mg(2+), and degrades plasmid fully in the presence of
CC       Mg(2+) and ATP, suggesting the helicase activity is required for
CC       complete degradation.
CC   -!- COFACTOR:
CC       Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC         Evidence={ECO:0000269|PubMed:21699496,
CC         ECO:0000269|PubMed:22521689};
CC   -!- SUBUNIT: Interacts with the CasA subunit of Cascade once Cascade
CC       has recognized target DNA. {ECO:0000269|PubMed:22521689}.
CC   -!- DOMAIN: Proteins of this family have an N-terminal nuclease domain
CC       and a C-terminal helicase/ATPase domain. In some CRISPR/Cas
CC       systems the domains are swapped, in others they are encoded
CC       separately.
CC   -!- DISRUPTION PHENOTYPE: Loss of plasmid silencing.
CC       {ECO:0000269|PubMed:18703739}.
CC   -!- SIMILARITY: In the N-terminal section; belongs to the CRISPR-
CC       associated nuclease Cas3-HD family. {ECO:0000305}.
CC   -!- SIMILARITY: In the central section; belongs to the CRISPR-
CC       associated helicase Cas3 family. {ECO:0000305}.
DR   EMBL; U29579; AAA69271.1; -; Genomic_DNA.
DR   EMBL; U00096; AAC75803.1; -; Genomic_DNA.
DR   EMBL; AP009048; BAE76838.1; -; Genomic_DNA.
DR   EMBL; Y07525; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR   PIR; E65057; E65057.
DR   RefSeq; NP_417241.1; NC_000913.3.
DR   RefSeq; WP_000433152.1; NZ_LN832404.1.
DR   BioGrid; 4260740; 78.
DR   IntAct; P38036; 7.
DR   STRING; 511145.b2761; -.
DR   PaxDb; P38036; -.
DR   PRIDE; P38036; -.
DR   EnsemblBacteria; AAC75803; AAC75803; b2761.
DR   EnsemblBacteria; BAE76838; BAE76838; BAE76838.
DR   GeneID; 947229; -.
DR   KEGG; ecj:JW2731; -.
DR   KEGG; eco:b2761; -.
DR   PATRIC; fig|1411691.4.peg.3976; -.
DR   EchoBASE; EB2516; -.
DR   eggNOG; ENOG4105DTT; Bacteria.
DR   eggNOG; COG1203; LUCA.
DR   HOGENOM; HOG000224183; -.
DR   InParanoid; P38036; -.
DR   KO; K07012; -.
DR   PhylomeDB; P38036; -.
DR   BioCyc; EcoCyc:EG12634-MONOMER; -.
DR   BioCyc; ECOL316407:JW2731-MONOMER; -.
DR   PRO; PR:P38036; -.
DR   Proteomes; UP000000318; Chromosome.
DR   Proteomes; UP000000625; Chromosome.
DR   GO; GO:0008296; F:3'-5'-exodeoxyribonuclease activity; IDA:EcoCyc.
DR   GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR   GO; GO:0033677; F:DNA/RNA helicase activity; IDA:EcoCyc.
DR   GO; GO:0097098; F:DNA/RNA hybrid annealing activity; IDA:EcoCyc.
DR   GO; GO:0003690; F:double-stranded DNA binding; IDA:EcoCyc.
DR   GO; GO:0000287; F:magnesium ion binding; EXP:EcoCyc.
DR   GO; GO:0000014; F:single-stranded DNA endodeoxyribonuclease activity; IDA:EcoCyc.
DR   GO; GO:0051607; P:defense response to virus; IDA:EcoCyc.
DR   GO; GO:0000737; P:DNA catabolic process, endonucleolytic; IDA:EcoCyc.
DR   GO; GO:0000738; P:DNA catabolic process, exonucleolytic; IDA:EcoCyc.
DR   Gene3D; 1.10.3210.30; -; 1.
DR   InterPro; IPR035011; Cas3.
DR   InterPro; IPR006483; CRISPR-assoc_Cas3_HD.
DR   InterPro; IPR038257; CRISPR-assoc_Cas3_HD_sf.
DR   InterPro; IPR011545; DEAD/DEAH_box_helicase_dom.
DR   InterPro; IPR014001; Helicase_ATP-bd.
DR   InterPro; IPR001650; Helicase_C.
DR   InterPro; IPR006474; Helicase_Cas3_CRISPR-ass_core.
DR   InterPro; IPR027417; P-loop_NTPase.
DR   PANTHER; PTHR24031:SF596; PTHR24031:SF596; 1.
DR   Pfam; PF00270; DEAD; 1.
DR   Pfam; PF18019; HD_6; 1.
DR   Pfam; PF00271; Helicase_C; 1.
DR   SMART; SM00487; DEXDc; 1.
DR   SMART; SM00490; HELICc; 1.
DR   SUPFAM; SSF52540; SSF52540; 1.
DR   TIGRFAMs; TIGR01587; cas3_core; 1.
DR   TIGRFAMs; TIGR01596; cas3_HD; 1.
DR   PROSITE; PS51643; HD_CAS3; 1.
DR   PROSITE; PS51192; HELICASE_ATP_BIND_1; 1.
DR   PROSITE; PS51194; HELICASE_CTER; 1.
PE   1: Evidence at protein level;
KW   Antiviral defense; ATP-binding; Complete proteome; DNA-binding;
KW   Endonuclease; Exonuclease; Helicase; Hydrolase; Magnesium;
KW   Metal-binding; Nuclease; Nucleotide-binding; Reference proteome.
FT   CHAIN         1    888       CRISPR-associated endonuclease/helicase
FT                                Cas3.
FT                                /FTId=PRO_0000207308.
FT   DOMAIN       20    231       HD Cas3-type. {ECO:0000255|PROSITE-
FT                                ProRule:PRU00974}.
FT   DOMAIN      301    504       Helicase ATP-binding.
FT                                {ECO:0000255|PROSITE-ProRule:PRU00541}.
FT   DOMAIN      556    735       Helicase C-terminal.
FT                                {ECO:0000255|PROSITE-ProRule:PRU00542}.
FT   NP_BIND     314    321       ATP. {ECO:0000255|PROSITE-
FT                                ProRule:PRU00541}.
FT   MOTIF       452    455       DEAH box.
FT   METAL        75     75       Magnesium. {ECO:0000255}.
FT   METAL       160    160       Magnesium. {ECO:0000255}.
FT   MUTAGEN      74     74       H->A: Loss of CRISPR immunity to lambda
FT                                DNA and of CRISPR-mediated plasmid
FT                                curing. {ECO:0000269|PubMed:21699496,
FT                                ECO:0000269|PubMed:22521689}.
FT   MUTAGEN      74     74       H->G: 75% R-loop formation.
FT                                {ECO:0000269|PubMed:21699496,
FT                                ECO:0000269|PubMed:22521689}.
FT   MUTAGEN      75     75       D->A: Loss of CRISPR immunity to lambda
FT                                DNA and of CRISPR-mediated plasmid
FT                                curing. {ECO:0000269|PubMed:21699496,
FT                                ECO:0000269|PubMed:22521689}.
FT   MUTAGEN      75     75       D->G: 10% R-loop formation.
FT                                {ECO:0000269|PubMed:21699496,
FT                                ECO:0000269|PubMed:22521689}.
FT   MUTAGEN      78     78       K->A: Loss of CRISPR immunity to lambda
FT                                DNA and of CRISPR-mediated plasmid
FT                                curing. {ECO:0000269|PubMed:21699496,
FT                                ECO:0000269|PubMed:22521689}.
FT   MUTAGEN      78     78       K->L: Less than 5% R-loop formation,
FT                                binds DNA normally. 100-fold greater
FT                                sensitivity to bacteriophage lambda. No
FT                                effect on R-loop formation; when
FT                                associated with L-320.
FT                                {ECO:0000269|PubMed:21699496,
FT                                ECO:0000269|PubMed:22521689}.
FT   MUTAGEN     317    318       GS->SY: No effect on R-loop formation.
FT                                Impaired R-loop unwinding.
FT                                {ECO:0000269|PubMed:21699496}.
FT   MUTAGEN     320    320       K->L: Double R-loop formation. 2-4 fold
FT                                decreased ATPase. No effect on R-loop
FT                                formation; when associated with L-78.
FT                                {ECO:0000269|PubMed:21699496,
FT                                ECO:0000269|PubMed:22521689}.
FT   MUTAGEN     320    320       K->N: Loss of CRISPR immunity to lambda
FT                                DNA and of CRISPR-mediated plasmid
FT                                curing. Nicks target plasmid DNA but does
FT                                not degrade it.
FT                                {ECO:0000269|PubMed:21699496,
FT                                ECO:0000269|PubMed:22521689}.
FT   MUTAGEN     452    452       D->N: Loss of CRISPR immunity to lambda
FT                                DNA and of CRISPR-mediated plasmid
FT                                curing. {ECO:0000269|PubMed:22521689}.
FT   MUTAGEN     455    455       H->L: Loss of R-loop unwinding.
FT                                {ECO:0000269|PubMed:21699496}.
FT   MUTAGEN     483    485       SAT->AAA: Retains CRISPR immunity to
FT                                lambda DNA and CRISPR-mediated plasmid
FT                                curing. {ECO:0000269|PubMed:22521689}.
FT   CONFLICT    118    118       G -> R (in Ref. 3; Y07525).
FT                                {ECO:0000305}.
FT   CONFLICT    334    335       QQ -> PL (in Ref. 3; Y07525).
FT                                {ECO:0000305}.
SQ   SEQUENCE   888 AA;  100545 MW;  4CA3F5371B1BF0F2 CRC64;
     MEPFKYICHY WGKSSKSLTK GNDIHLLIYH CLDVAAVADC WWDQSVVLQN TFCRNEMLSK
     QRVKAWLLFF IALHDIGKFD IRFQYKSAES WLKLNPATPS LNGPSTQMCR KFNHGAAGLY
     WFNQDSLSEQ SLGDFFSFFD AAPHPYESWF PWVEAVTGHH GFILHSQDQD KSRWEMPASL
     ASYAAQDKQA REEWISVLEA LFLTPAGLSI NDIPPDCSSL LAGFCSLADW LGSWTTTNTF
     LFNEDAPSDI NALRTYFQDR QQDASRVLEL SGLVSNKRCY EGVHALLDNG YQPRQLQVLV
     DALPVAPGLT VIEAPTGSGK TETALAYAWK LIDQQIADSV IFALPTQATA NAMLTRMEAS
     ASHLFSSPNL ILAHGNSRFN HLFQSIKSRA ITEQGQEEAW VQCCQWLSQS NKKVFLGQIG
     VCTIDQVLIS VLPVKHRFIR GLGIGRSVLI VDEVHAYDTY MNGLLEAVLK AQADVGGSVI
     LLSATLPMKQ KQKLLDTYGL HTDPVENNSA YPLINWRGVN GAQRFDLLAH PEQLPPRFSI
     QPEPICLADM LPDLTMLERM IAAANAGAQV CLICNLVDVA QVCYQRLKEL NNTQVDIDLF
     HARFTLNDRR EKENRVISNF GKNGKRNVGR ILVATQVVEQ SLDVDFDWLI TQHCPADLLF
     QRLGRLHRHH RKYRPAGFEI PVATILLPDG EGYGRHEHIY SNVRVMWRTQ QHIEELNGAS
     LFFPDAYRQW LDSIYDDAEM DEPEWVGNGM DKFESAECEK RFKARKVLQW AEEYSLQDND
     ETILAVTRDG EMSLPLLPYV QTSSGKQLLD GQVYEDLSHE QQYEALALNR VNVPFTWKRS
     FSEVVDEDGL LWLEGKQNLD GWVWQGNSIV ITYTGDEGMT RVIPANPK
//
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