CAS10_STRTR
ID CAS10_STRTR Reviewed; 758 AA.
AC A0A0A7HFE1;
DT 16-JAN-2019, integrated into UniProtKB/Swiss-Prot.
DT 04-MAR-2015, sequence version 1.
DT 25-MAY-2022, entry version 18.
DE RecName: Full=CRISPR system single-strand-specific deoxyribonuclease Cas10/Csm1 (subtype III-A);
DE Short=ssDNase Cas10;
DE EC=3.1.-.- {ECO:0000269|PubMed:27105119};
DE AltName: Full=Cyclic oligoadenylate synthase {ECO:0000305|PubMed:28663439};
DE EC=2.7.7.- {ECO:0000269|PubMed:28663439};
DE AltName: Full=StCas10 {ECO:0000303|PubMed:27105119};
GN Name=cas10 {ECO:0000303|PubMed:25458845}; Synonyms=csm1 {ECO:0000305};
OS Streptococcus thermophilus.
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Streptococcus.
OX NCBI_TaxID=1308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION IN PHAGE RESISTANCE, TARGETS
RP SSRNA, SUBUNIT, AND ANTIVIRAL DEFENSE.
RC STRAIN=DGCC8004;
RX PubMed=25458845; DOI=10.1016/j.molcel.2014.09.027;
RA Tamulaitis G., Kazlauskiene M., Manakova E., Venclovas C., Nwokeoji A.O.,
RA Dickman M.J., Horvath P., Siksnys V.;
RT "Programmable RNA shredding by the type III-A CRISPR-Cas system of
RT Streptococcus thermophilus.";
RL Mol. Cell 56:506-517(2014).
RN [2]
RP FUNCTION IN SSDNASE ACTIVITY, COFACTOR, ACTIVITY REGULATION, SUBUNIT,
RP DOMAIN, AND MUTAGENESIS OF ASP-16 AND 575-ASP-ASP-576.
RC STRAIN=DGCC8004;
RX PubMed=27105119; DOI=10.1016/j.molcel.2016.03.024;
RA Kazlauskiene M., Tamulaitis G., Kostiuk G., Venclovas C., Siksnys V.;
RT "Spatiotemporal control of type III-A CRISPR-Cas immunity: coupling DNA
RT degradation with the target RNA recognition.";
RL Mol. Cell 62:295-306(2016).
RN [3]
RP FUNCTION IN COA FORMATION, COFACTOR, SUBUNIT, DOMAIN, AND MUTAGENESIS OF
RP ASP-16 AND 575-ASP-ASP-576.
RC STRAIN=DGCC8004;
RX PubMed=28663439; DOI=10.1126/science.aao0100;
RA Kazlauskiene M., Kostiuk G., Venclovas C., Tamulaitis G., Siksnys V.;
RT "A cyclic oligonucleotide signaling pathway in type III CRISPR-Cas
RT systems.";
RL Science 357:605-609(2017).
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 spacers, sequences complementary to
CC antecedent mobile elements, and target invading nucleic acids. CRISPR
CC clusters are transcribed and processed into CRISPR RNA (crRNA). The
CC type III-A Csm effector complex binds crRNA and acts as a crRNA-guided
CC RNase, DNase and cyclic oligoadenylate synthase; binding of target RNA
CC cognate to the crRNA is required for all activities. In a heterologous
CC host this Csm effector complex restricts ssRNA phage MS2, suggesting it
CC may target RNA viruses in vivo. {ECO:0000269|PubMed:25458845}.
CC -!- FUNCTION: Csm functions as a non-specific ssDNase. Base-pairing between
CC crRNA and target RNA to form a ternary Csm complex activates a ssDNase
CC activity; target RNA cleavage suppresses the ssDNase, a temporal
CC control that prevents uncontrolled DNA degradation. Viral RNA
CC transcripts probably tether the Csm complex to the viral genome,
CC recruiting Cas10 ssDNA activity which is able to degrade DNA in the
CC transcription bubble, spatially controlling the DNase activity.
CC {ECO:0000269|PubMed:27105119}.
CC -!- FUNCTION: This subunit has a weak ssDNase activity that is dramatically
CC activated by the ternary Csm effector complex (the crRNA, Cas proteins
CC and a cognate target ssRNA). Target RNA and ssDNA are cleaved
CC simultaneously, although RNase activity (of Csm3) is much faster. RNA
CC cleavage by Csm3 is not required for ssDNase activity as Csm complex
CC with inactive Csm3 still has ssDNase activity; however as the cleaved
CC target RNA products dissociate away ssDNase activity decreases. Self-
CC recognition, with subsequent repression of the ssDNase activity, occurs
CC when the 5' handle of the crRNA bases pairs with the 3' flanking
CC sequence of the target RNA (which would occur if the CRISPR locus were
CC transcribed as an anti-pre-crRNA). This protein has low activity on
CC dsDNA which is not stimulated by the Csm complex.
CC {ECO:0000269|PubMed:27105119}.
CC -!- FUNCTION: This subunit is a single-strand-specific deoxyribonuclease
CC (ssDNase) which digests both linear and circular ssDNA; it has both
CC exo- and endonuclease activity. {ECO:0000250|UniProtKB:B6YWB8}.
CC -!- FUNCTION: When associated with the ternary Csm effector complex (the
CC crRNA, Cas proteins and a cognate target ssRNA) synthesizes cyclic
CC oligoadenylates (cOA) from ATP, producing cyclic triadenylate (cA3) up
CC to cyclic hexaadenylate (cA6), which is the active cOA. The enzyme is
CC also able to cyclize pppA3 up to pppA6. cOAs are second messengers that
CC induce an antiviral state important for defense against invading
CC nucleic acids. Synthesis of cOA can occur with AMP plus ATP, 2'dATP or
CC 3'dATP (but no other nucleotides), and requires a free 3'-OH ribose
CC moiety. {ECO:0000269|PubMed:28663439}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=6 ATP = cyclic hexaadenylate + 6 diphosphate;
CC Xref=Rhea:RHEA:58276, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:142456; Evidence={ECO:0000269|PubMed:28663439};
CC -!- COFACTOR:
CC Name=a divalent metal cation; Xref=ChEBI:CHEBI:60240;
CC Evidence={ECO:0000269|PubMed:27105119, ECO:0000269|PubMed:28663439};
CC Note=Degradation of ssDNA requires Mn(2+), Co(2+) or Ni(2+); Mg(2+),
CC Ca(2+) or Zn(2+) do not activate ssDNase activity (PubMed:27105119).
CC Formation of cOA requires Mn(2+), Co(2+) or Zn(2+), Mg(2+) is not as
CC efficient (PubMed:28663439). {ECO:0000269|PubMed:27105119,
CC ECO:0000269|PubMed:28663439};
CC -!- ACTIVITY REGULATION: ssDNase activity is activated by target RNA
CC binding to the Csm-crRNA complex and is inhibited by EDTA.
CC {ECO:0000269|PubMed:27105119}.
CC -!- SUBUNIT: Part of the Csm effector complex that includes at least
CC Cas10(1), Csm2(3), Csm3(5), Csm4(1), Csm5(1) and mature crRNA
CC (PubMed:25458845, PubMed:27105119, PubMed:28663439). The Csm complex is
CC elongated and slightly twisted with a maximal length of 215 Angstroms
CC and a diameter of 75-80 Angstroms (PubMed:25458845). It has been
CC modeled to have a central protein filamant of Csm3 subunits along which
CC the dsRNA helix of paired crRNA and target RNA binds. The filament is
CC capped at one end by Cas10 and Csm4 and at the other end by Csm5; ssDNA
CC is thought to bind to the N-terminal HD domain of Cas10 (Probable). Csm
CC with a precursor crRNA does not include Csm5, while Cas6, the enzyme
CC probably involved in pre-crRNA processing, is found associated with a
CC subset of the Csm complex (PubMed:25458845).
CC {ECO:0000269|PubMed:25458845, ECO:0000269|PubMed:27105119,
CC ECO:0000269|PubMed:28663439, ECO:0000305|PubMed:25458845,
CC ECO:0000305|PubMed:27105119}.
CC -!- DOMAIN: The N-terminal HD domain has ssDNase activity
CC (PubMed:27105119). The C-terminal GGDEF domain has the cOA synthesis
CC activity (PubMed:28663439). {ECO:0000269|PubMed:27105119,
CC ECO:0000269|PubMed:28663439}.
CC -!- MISCELLANEOUS: Encoded in a type III-A CRISPR locus.
CC {ECO:0000269|PubMed:25458845}.
CC -!- SIMILARITY: Belongs to the CRISPR-associated Cas10/Csm1 family.
CC {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; KM222358; AIZ03604.1; -; Genomic_DNA.
DR RefSeq; WP_014621547.1; NZ_CP049053.1.
DR PDB; 6NUD; EM; 3.50 A; J=1-758.
DR PDB; 6NUE; EM; 3.30 A; J=1-758.
DR PDBsum; 6NUD; -.
DR PDBsum; 6NUE; -.
DR AlphaFoldDB; A0A0A7HFE1; -.
DR SMR; A0A0A7HFE1; -.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0004519; F:endonuclease activity; IEA:UniProtKB-KW.
DR GO; GO:0004527; F:exonuclease activity; IEA:UniProtKB-KW.
DR GO; GO:0003723; F:RNA binding; IEA:UniProtKB-KW.
DR GO; GO:0016740; F:transferase activity; IEA:UniProtKB-KW.
DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-KW.
DR CDD; cd09680; Cas10_III; 1.
DR Gene3D; 3.30.70.270; -; 1.
DR InterPro; IPR013408; Cas10/Csm1.
DR InterPro; IPR041062; Csm1_B.
DR InterPro; IPR000160; GGDEF_dom.
DR InterPro; IPR006674; HD_domain.
DR InterPro; IPR043128; Rev_trsase/Diguanyl_cyclase.
DR Pfam; PF18211; Csm1_B; 1.
DR Pfam; PF01966; HD; 1.
DR TIGRFAMs; TIGR02578; cas_TM1811_Csm1; 1.
DR PROSITE; PS50887; GGDEF; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antiviral defense; ATP-binding; Endonuclease; Exonuclease;
KW Hydrolase; Nuclease; Nucleotide-binding; RNA-binding; Transferase.
FT CHAIN 1..758
FT /note="CRISPR system single-strand-specific
FT deoxyribonuclease Cas10/Csm1 (subtype III-A)"
FT /id="PRO_0000446112"
FT DOMAIN 509..647
FT /note="GGDEF"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00095,
FT ECO:0000305|PubMed:27105119"
FT REGION 1..82
FT /note="HD domain"
FT /evidence="ECO:0000305|PubMed:27105119"
FT MUTAGEN 16
FT /note="D->A: Dramatically decreased ssDNase activity. Wild-
FT type synthesis of cOA."
FT /evidence="ECO:0000269|PubMed:27105119,
FT ECO:0000269|PubMed:28663439"
FT MUTAGEN 575..576
FT /note="DD->AA: Wild-type ssDNase activity. No synthesis of
FT cOA."
FT /evidence="ECO:0000269|PubMed:27105119,
FT ECO:0000269|PubMed:28663439"
FT HELIX 6..13
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 15..17
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 18..24
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 31..42
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 47..53
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 55..58
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 73..80
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 106..108
FT /evidence="ECO:0007829|PDB:6NUD"
FT HELIX 111..113
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 114..116
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 130..132
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 138..141
FT /evidence="ECO:0007829|PDB:6NUD"
FT HELIX 153..158
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 174..177
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 178..180
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 181..183
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 185..187
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 190..192
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 193..197
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 198..218
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 225..227
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 235..237
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 241..247
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 252..256
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 261..266
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 267..289
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 293..295
FT /evidence="ECO:0007829|PDB:6NUD"
FT STRAND 306..308
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 312..325
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 328..331
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 341..346
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 348..350
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 351..354
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 358..375
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 376..378
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 382..384
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 385..388
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 400..403
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 412..416
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 419..421
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 422..427
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 428..431
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 433..436
FT /evidence="ECO:0007829|PDB:6NUD"
FT STRAND 439..443
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 446..448
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 452..456
FT /evidence="ECO:0007829|PDB:6NUD"
FT STRAND 472..474
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 496..499
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 503..505
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 511..514
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 523..525
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 533..535
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 540..554
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 557..561
FT /evidence="ECO:0007829|PDB:6NUE"
FT TURN 562..564
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 565..571
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 574..576
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 578..581
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 583..601
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 610..614
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 620..633
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 634..636
FT /evidence="ECO:0007829|PDB:6NUD"
FT STRAND 637..640
FT /evidence="ECO:0007829|PDB:6NUD"
FT STRAND 645..648
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 654..657
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 660..663
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 665..671
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 673..675
FT /evidence="ECO:0007829|PDB:6NUD"
FT TURN 677..680
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 683..691
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 694..697
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 698..713
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 718..721
FT /evidence="ECO:0007829|PDB:6NUE"
FT STRAND 723..725
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 726..732
FT /evidence="ECO:0007829|PDB:6NUE"
FT HELIX 738..754
FT /evidence="ECO:0007829|PDB:6NUE"
SQ SEQUENCE 758 AA; 86822 MW; E3E3E4B04AEAF9B6 CRC64;
MKKEKIDLFY GALLHDIGKV IQRATGERKK HALVGADWFD EIADNQVISD QIRYHMANYQ
SDKLGNDHLA YITYIADNIA SGVDRRQSNE ESDEDASAKI WDTYTNQADI FNVFGAQTDK
RYFKPTVLNL KSKPNFASAT YEPFSKGDYA AIATRIKNEL AEFEFNQAQI DSLLNLFEAI
LSFVPSSTNS KEIADISLAE HSRLTAAFAL AIYDYLEDKG RHNYKEDLFT KASAFYEEEA
FLLASFDLSG IQDFIYNIAT SGAAKQLKAR SLYLDFMSEY IADSLLDKLG LNRANLLYVG
GGHAYFVLAN TEKTVETLVQ FEKDFNQFLL ANFQTRLYVA FGWGSFAAKD IMSELNSPES
YRQIYQKASR MISEKKISRY DYRTLMLLNR GGKSSERECE ICHSVENLVS YHDQKVCDIC
RGLYQFSKEI AHDHFIITEN EGLPIGPNAC LKGVAFEKLS QESFSRVYVK NDYKAGTIKA
THVFVGDYQC DEIHKYAALS KNEDGLGIKR LAVVRLDVDD LGAAFMAGFS RQGNGQYSTL
SRSATFSRSM SLFFKVYINQ FASDKKLSII YAGGDDVFAI GSWQDIIAFT VELRQNFIKW
TNGKLTLSAG IGLFADKTPI SLMAHQTGEL EEAAKGNEKD SISLFSSDYT FKFDRFITNV
YDDKLEQIRY FFNHQDERGK NFIYKLIELL RNYESEEKMN VARLAYYLTR LEELTDKDER
DKFKQFKKLF FKWYTNNESD RKEAELALLL YVYEIRKD
