CAS4_SACS2
ID CAS4_SACS2 Reviewed; 202 AA.
AC Q97TX9;
DT 01-MAY-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2001, sequence version 1.
DT 03-AUG-2022, entry version 90.
DE RecName: Full=CRISPR-associated exonuclease Cas4;
DE EC=3.1.12.1 {ECO:0000269|PubMed:23056615, ECO:0000269|PubMed:24171432};
GN Name=cas4; OrderedLocusNames=SSO0001;
OS Saccharolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
OS (Sulfolobus solfataricus).
OC Archaea; Crenarchaeota; Thermoprotei; Sulfolobales; Sulfolobaceae;
OC Saccharolobus.
OX NCBI_TaxID=273057;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 35092 / DSM 1617 / JCM 11322 / P2;
RX PubMed=11427726; DOI=10.1073/pnas.141222098;
RA She Q., Singh R.K., Confalonieri F., Zivanovic Y., Allard G., Awayez M.J.,
RA Chan-Weiher C.C.-Y., Clausen I.G., Curtis B.A., De Moors A., Erauso G.,
RA Fletcher C., Gordon P.M.K., Heikamp-de Jong I., Jeffries A.C., Kozera C.J.,
RA Medina N., Peng X., Thi-Ngoc H.P., Redder P., Schenk M.E., Theriault C.,
RA Tolstrup N., Charlebois R.L., Doolittle W.F., Duguet M., Gaasterland T.,
RA Garrett R.A., Ragan M.A., Sensen C.W., Van der Oost J.;
RT "The complete genome of the crenarchaeon Sulfolobus solfataricus P2.";
RL Proc. Natl. Acad. Sci. U.S.A. 98:7835-7840(2001).
RN [2]
RP FUNCTION AS AN EXONUCLEASE, CATALYTIC ACTIVITY, COFACTOR, SUBUNIT, AND
RP MUTAGENESIS OF ASP-99.
RC STRAIN=ATCC 35092 / DSM 1617 / JCM 11322 / P2;
RX PubMed=23056615; DOI=10.1371/journal.pone.0047232;
RA Zhang J., Kasciukovic T., White M.F.;
RT "The CRISPR associated protein Cas4 is a 5' to 3' DNA exonuclease with an
RT iron-sulfur cluster.";
RL PLoS ONE 7:E47232-E47232(2012).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.35 ANGSTROMS) IN COMPLEX WITH IRON-SULFUR (4FE-4S)
RP AND MANGANESE, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL
RP PROPERTIES, DNA-BINDING, SUBUNIT, AND MUTAGENESIS OF CYS-32; ASP-99;
RP LYS-115; TYR-148; CYS-188; CYS-191 AND CYS-197.
RX PubMed=24171432; DOI=10.1021/ja408729b;
RA Lemak S., Beloglazova N., Nocek B., Skarina T., Flick R., Brown G.,
RA Popovic A., Joachimiak A., Savchenko A., Yakunin A.F.;
RT "Toroidal structure and DNA cleavage by the CRISPR-associated [4Fe-4S]
RT cluster containing Cas4 nuclease SSO0001 from Sulfolobus solfataricus.";
RL J. Am. Chem. Soc. 135:17476-17487(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 sequences complementary to
CC antecedent mobile elements and target invading nucleic acids. CRISPR
CC clusters are transcribed and processed into CRISPR RNA (crRNA) (By
CC similarity). This protein is a 5' to 3' partially processive
CC exonuclease that cleaves off single mononucleotides. Has a marked
CC preference for ssDNA, although in vitro it also acts on dsDNA and
CC ssRNA. Has low endonuclease activity with circular ssDNA. Binds ssDNA
CC and can unwind dsDNA; unwinding does not require ATP. {ECO:0000250,
CC ECO:0000269|PubMed:23056615, ECO:0000269|PubMed:24171432}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=exonucleolytic cleavage in the 5'- to 3'-direction to yield
CC nucleoside 3'-phosphates.; EC=3.1.12.1;
CC Evidence={ECO:0000269|PubMed:23056615, ECO:0000269|PubMed:24171432};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:23056615, ECO:0000269|PubMed:24171432};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:23056615, ECO:0000269|PubMed:24171432};
CC Name=Cu(2+); Xref=ChEBI:CHEBI:29036;
CC Evidence={ECO:0000269|PubMed:23056615, ECO:0000269|PubMed:24171432};
CC Note=Mg(2+) or Mn(2+) required for ssDNA cleavage activity. Can also
CC utilise Cu(2+). {ECO:0000269|PubMed:23056615,
CC ECO:0000269|PubMed:24171432};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000269|PubMed:23056615, ECO:0000269|PubMed:24171432,
CC ECO:0000305};
CC Note=Binds 1 [4Fe-4S] cluster per subunit (PubMed:24171432). It may be
CC important for protein stability, since mutation of the Cys that bind
CC the cofactor leads to a colorless, insoluble protein.
CC {ECO:0000269|PubMed:23056615, ECO:0000269|PubMed:24171432,
CC ECO:0000305};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 9-10. {ECO:0000269|PubMed:24171432};
CC Temperature dependence:
CC Optimum temperature is 65-70 degrees Celsius.
CC {ECO:0000269|PubMed:24171432};
CC -!- SUBUNIT: Homodecamer, formed by the assembly of dimers into a decameric
CC toroid, with the active sites oriented towards the central tunnel.
CC {ECO:0000269|PubMed:23056615, ECO:0000269|PubMed:24171432}.
CC -!- SIMILARITY: Belongs to the CRISPR-associated exonuclease Cas4 family.
CC {ECO:0000305}.
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DR EMBL; AE006641; AAK43340.1; -; Genomic_DNA.
DR PIR; A90139; A90139.
DR RefSeq; WP_009989652.1; NC_002754.1.
DR PDB; 4IC1; X-ray; 2.35 A; A/B/C/D/F/G/H/I/J/K=1-202.
DR PDBsum; 4IC1; -.
DR AlphaFoldDB; Q97TX9; -.
DR SMR; Q97TX9; -.
DR STRING; 273057.SSO0001; -.
DR EnsemblBacteria; AAK43340; AAK43340; SSO0001.
DR GeneID; 44128964; -.
DR KEGG; sso:SSO0001; -.
DR PATRIC; fig|273057.12.peg.2; -.
DR eggNOG; arCOG00790; Archaea.
DR HOGENOM; CLU_106570_0_0_2; -.
DR InParanoid; Q97TX9; -.
DR OMA; DWECRYC; -.
DR PhylomeDB; Q97TX9; -.
DR BRENDA; 3.1.12.1; 6163.
DR Proteomes; UP000001974; Chromosome.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IDA:UniProtKB.
DR GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR GO; GO:0030145; F:manganese ion binding; IDA:UniProtKB.
DR GO; GO:0045145; F:single-stranded DNA 5'-3' exodeoxyribonuclease activity; IDA:UniProtKB.
DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-KW.
DR GO; GO:0000738; P:DNA catabolic process, exonucleolytic; IDA:UniProtKB.
DR GO; GO:0032508; P:DNA duplex unwinding; IDA:UniProtKB.
DR Gene3D; 3.90.320.10; -; 1.
DR InterPro; IPR013343; CRISPR-assoc_prot_Cas4.
DR InterPro; IPR011604; PDDEXK-like_dom_sf.
DR InterPro; IPR038726; PDDEXK_AddAB-type.
DR Pfam; PF12705; PDDEXK_1; 1.
DR TIGRFAMs; TIGR00372; cas4; 1.
PE 1: Evidence at protein level;
KW 3D-structure; 4Fe-4S; Antiviral defense; DNA-binding; Exonuclease;
KW Hydrolase; Iron; Iron-sulfur; Manganese; Metal-binding; Nuclease;
KW Reference proteome.
FT CHAIN 1..202
FT /note="CRISPR-associated exonuclease Cas4"
FT /id="PRO_0000422224"
FT BINDING 32
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000269|PubMed:24171432"
FT BINDING 62
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:24171432"
FT BINDING 99
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:24171432"
FT BINDING 113
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:24171432"
FT BINDING 114
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000269|PubMed:24171432"
FT BINDING 188
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000269|PubMed:24171432"
FT BINDING 191
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000269|PubMed:24171432"
FT BINDING 197
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000269|PubMed:24171432"
FT MUTAGEN 32
FT /note="C->A: Abolishes iron-sulfur (4Fe-4S) binding and
FT exonuclease activity; when associated with A-188; A-191 and
FT A-197."
FT /evidence="ECO:0000269|PubMed:24171432"
FT MUTAGEN 99
FT /note="D->A: Abolishes nuclease activity."
FT /evidence="ECO:0000269|PubMed:23056615,
FT ECO:0000269|PubMed:24171432"
FT MUTAGEN 113
FT /note="E->A: Impairs exonuclease activity."
FT /evidence="ECO:0000269|PubMed:24171432"
FT MUTAGEN 115
FT /note="K->A: Abolishes exonuclease activity."
FT /evidence="ECO:0000269|PubMed:24171432"
FT MUTAGEN 148
FT /note="Y->A: Nearly abolishes exonuclease activity."
FT /evidence="ECO:0000269|PubMed:24171432"
FT MUTAGEN 188
FT /note="C->A: Abolishes iron-sulfur (4Fe-4S) binding and
FT exonuclease activity; when associated with A-32; A-191 and
FT A-197."
FT /evidence="ECO:0000269|PubMed:24171432"
FT MUTAGEN 191
FT /note="C->A: Abolishes iron-sulfur (4Fe-4S) binding and
FT exonuclease activity; when associated with A-32; A-188 and
FT A-197."
FT /evidence="ECO:0000269|PubMed:24171432"
FT MUTAGEN 197
FT /note="C->A: Abolishes iron-sulfur (4Fe-4S) binding and
FT exonuclease activity; when associated with A-32; A-188 and
FT A-191."
FT /evidence="ECO:0000269|PubMed:24171432"
FT HELIX 1..16
FT /evidence="ECO:0007829|PDB:4IC1"
FT STRAND 23..25
FT /evidence="ECO:0007829|PDB:4IC1"
FT HELIX 26..29
FT /evidence="ECO:0007829|PDB:4IC1"
FT HELIX 33..41
FT /evidence="ECO:0007829|PDB:4IC1"
FT HELIX 43..46
FT /evidence="ECO:0007829|PDB:4IC1"
FT HELIX 47..51
FT /evidence="ECO:0007829|PDB:4IC1"
FT HELIX 53..72
FT /evidence="ECO:0007829|PDB:4IC1"
FT STRAND 75..87
FT /evidence="ECO:0007829|PDB:4IC1"
FT STRAND 90..102
FT /evidence="ECO:0007829|PDB:4IC1"
FT STRAND 105..107
FT /evidence="ECO:0007829|PDB:4IC1"
FT STRAND 110..115
FT /evidence="ECO:0007829|PDB:4IC1"
FT HELIX 126..139
FT /evidence="ECO:0007829|PDB:4IC1"
FT STRAND 142..150
FT /evidence="ECO:0007829|PDB:4IC1"
FT STRAND 153..159
FT /evidence="ECO:0007829|PDB:4IC1"
FT HELIX 165..176
FT /evidence="ECO:0007829|PDB:4IC1"
FT STRAND 180..184
FT /evidence="ECO:0007829|PDB:4IC1"
FT HELIX 188..190
FT /evidence="ECO:0007829|PDB:4IC1"
FT HELIX 194..196
FT /evidence="ECO:0007829|PDB:4IC1"
SQ SEQUENCE 202 AA; 23239 MW; C1E8868ACC093382 CRC64;
MITEFLLKKK LEEHLSHVKE ENTIYVTDLV RCPRRVRYES EYKELAISQV YAPSAILGDI
LHLGLESVLK GNFNAETEVE TLREINVGGK VYKIKGRADA IIRNDNGKSI VIEIKTSRSD
KGLPLIHHKM QLQIYLWLFS AEKGILVYIT PDRIAEYEIN EPLDEATIVR LAEDTIMLQN
SPRFNWECKY CIFSVICPAK LT
