CSA1_SACS2
ID CSA1_SACS2 Reviewed; 291 AA.
AC Q97YD4;
DT 01-MAY-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2001, sequence version 1.
DT 03-AUG-2022, entry version 83.
DE RecName: Full=CRISPR-associated exonuclease Csa1;
GN Name=csa1; OrderedLocusNames=SSO1391;
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 POSSIBLE COFACTOR.
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 FUNCTION, COFACTOR, AND MUTAGENESIS OF GLU-195.
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).
RN [4]
RP FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, SUBUNIT, AND MUTAGENESIS
RP OF CYS-52; TYR-59; HIS-80; ASP-184; GLU-195; LYS-197; TYR-201; TYR-212;
RP GLU-217; CYS-230; CYS-243; ARG-260; CYS-283 AND CYS-289.
RX PubMed=25200083; DOI=10.1093/nar/gku797;
RA Lemak S., Nocek B., Beloglazova N., Skarina T., Flick R., Brown G.,
RA Joachimiak A., Savchenko A., Yakunin A.F.;
RT "The CRISPR-associated Cas4 protein Pcal_0546 from Pyrobaculum calidifontis
RT contains a [2Fe-2S] cluster: crystal structure and nuclease activity.";
RL Nucleic Acids Res. 42:11144-11155(2014).
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). A ssDNA exonuclease that has both 5' to 3' and 3' to 5'
CC activity, yielding 5'-OH and 3'-phosphate groups. Has Mn(2+)-dependent
CC endonuclease activity on circular ssDNA. Can unwind dsDNA; unwinding
CC does not require ATP. {ECO:0000250, ECO:0000269|PubMed:24171432,
CC ECO:0000269|PubMed:25200083}.
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:24171432, ECO:0000269|PubMed:25200083};
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:24171432, ECO:0000269|PubMed:25200083};
CC Note=Mn(2+) required for nuclease activity. Can also utilize Co(2+) and
CC to a lesser extent Mg(2+) or Ni(2+). {ECO:0000269|PubMed:24171432,
CC ECO:0000269|PubMed:25200083};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000250|UniProtKB:Q97TX9, ECO:0000303|PubMed:23056615,
CC ECO:0000303|PubMed:24171432, ECO:0000303|PubMed:25200083};
CC Note=Binds 1 [4Fe-4S] cluster per subunit. Not required for nuclease
CC activity, since mutation of the Cys residues leads to a colorless but
CC active protein. {ECO:0000250|UniProtKB:Q97TX9,
CC ECO:0000303|PubMed:23056615, ECO:0000303|PubMed:24171432,
CC ECO:0000303|PubMed:25200083};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 7-9. {ECO:0000269|PubMed:25200083};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:25200083}.
CC -!- SIMILARITY: Belongs to the CRISPR-associated protein Csa1 family.
CC {ECO:0000305}.
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DR EMBL; AE006641; AAK41627.1; -; Genomic_DNA.
DR PIR; D90296; D90296.
DR AlphaFoldDB; Q97YD4; -.
DR STRING; 273057.SSO1391; -.
DR PRIDE; Q97YD4; -.
DR EnsemblBacteria; AAK41627; AAK41627; SSO1391.
DR KEGG; sso:SSO1391; -.
DR PATRIC; fig|273057.12.peg.1407; -.
DR eggNOG; arCOG04195; Archaea.
DR HOGENOM; CLU_905009_0_0_2; -.
DR InParanoid; Q97YD4; -.
DR OMA; GYLCYVN; -.
DR PhylomeDB; Q97YD4; -.
DR Proteomes; UP000001974; Chromosome.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008310; F:single-stranded DNA 3'-5' exodeoxyribonuclease activity; IDA:UniProtKB.
DR GO; GO:0045145; F:single-stranded DNA 5'-3' exodeoxyribonuclease activity; IDA:UniProtKB.
DR GO; GO:0000014; F:single-stranded DNA endodeoxyribonuclease activity; IDA:UniProtKB.
DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-KW.
DR GO; GO:0032508; P:DNA duplex unwinding; IDA:UniProtKB.
DR Gene3D; 3.90.320.10; -; 1.
DR InterPro; IPR009260; CRISPR-ass_Csa1.
DR InterPro; IPR011604; PDDEXK-like_dom_sf.
DR Pfam; PF06023; Csa1; 1.
DR PIRSF; PIRSF009226; UCP009226; 1.
DR TIGRFAMs; TIGR01896; cas_AF1879; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Antiviral defense; Exonuclease; Hydrolase; Iron; Iron-sulfur;
KW Manganese; Metal-binding; Nuclease; Reference proteome.
FT CHAIN 1..291
FT /note="CRISPR-associated exonuclease Csa1"
FT /id="PRO_0000422231"
FT BINDING 52
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:Q97TX9"
FT BINDING 279
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:Q97TX9"
FT BINDING 283
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:Q97TX9"
FT BINDING 289
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000250|UniProtKB:Q97TX9"
FT MUTAGEN 52
FT /note="C->A: Nearly wild-type protein."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 59
FT /note="Y->A: No effect."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 80
FT /note="H->A: No exonuclease, endonuclease, or DNA unwinding
FT activity."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 184
FT /note="D->A: No exonuclease, endonuclease, or DNA unwinding
FT activity."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 195
FT /note="E->A: No exonuclease, endonuclease, or DNA unwinding
FT activity."
FT /evidence="ECO:0000269|PubMed:24171432,
FT ECO:0000269|PubMed:25200083"
FT MUTAGEN 197
FT /note="K->A: No exonuclease, endonuclease, or DNA unwinding
FT activity."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 201
FT /note="Y->A: No effect."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 212
FT /note="Y->A: No exonuclease, endonuclease, or DNA unwinding
FT activity."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 217
FT /note="E->A: Reduced exonuclease and DNA unwinding, no
FT endonuclease activity."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 230
FT /note="C->A: No effect."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 243
FT /note="C->A: No effect."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 260
FT /note="R->A: Wild-type exonuclease, reduced DNA unwinding,
FT no endonuclease activity."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 283
FT /note="C->A: Wild-type exonuclease and DNA unwinding,
FT reduced endonuclease activity."
FT /evidence="ECO:0000269|PubMed:25200083"
FT MUTAGEN 289
FT /note="C->A: Wild-type exonuclease and DNA unwinding,
FT reduced endonuclease activity."
FT /evidence="ECO:0000269|PubMed:25200083"
SQ SEQUENCE 291 AA; 33491 MW; BE68EC7DC4F5AC0B CRC64;
MFFTHSDMLL LSKRIKKLPK NVDEELRGWN WSEPPVYTRS LSQVSISEMV YCSTLRNVYL
KVKGFRGEIG RQILQGSLIH TIYAIGIEAI KRFIYSRESI DGSTLRTLMG DEFYSLLKDL
REEEGIYAKV LWDHITNIYS AELDRVRSKF TNLTRDSLVS QVVPFYVEFP VDGSLLGLTN
LRVDAFIPHL PLIAEMKTGK YRYTHELSLA GYALAIESQY EIPIDFGYLC YVTVTEKEVK
NNCKLIPISD SLRSEFLDMR DKAQDIMDKG VDPGIAKDCE SDCMFYKVCH P
