CAS6_PSEAB
ID CAS6_PSEAB Reviewed; 187 AA.
AC Q02MM2;
DT 13-JUN-2012, integrated into UniProtKB/Swiss-Prot.
DT 13-JUN-2012, sequence version 2.
DT 03-AUG-2022, entry version 68.
DE RecName: Full=CRISPR-associated endonuclease Cas6/Csy4;
DE EC=3.1.-.-;
GN Name=cas6f; Synonyms=csy4; OrderedLocusNames=PA14_33300;
OS Pseudomonas aeruginosa (strain UCBPP-PA14).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=208963;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=UCBPP-PA14;
RX PubMed=17038190; DOI=10.1186/gb-2006-7-10-r90;
RA Lee D.G., Urbach J.M., Wu G., Liberati N.T., Feinbaum R.L., Miyata S.,
RA Diggins L.T., He J., Saucier M., Deziel E., Friedman L., Li L., Grills G.,
RA Montgomery K., Kucherlapati R., Rahme L.G., Ausubel F.M.;
RT "Genomic analysis reveals that Pseudomonas aeruginosa virulence is
RT combinatorial.";
RL Genome Biol. 7:R90.1-R90.14(2006).
RN [2]
RP FUNCTION IN CRRNA FORMATION, FUNCTION IN INHIBITION OF BIOFILM FORMATION,
RP DISRUPTION PHENOTYPE, AND MUTAGENESIS OF HIS-29.
RC STRAIN=UCBPP-PA14;
RX PubMed=21398535; DOI=10.1128/jb.01411-10;
RA Cady K.C., O'Toole G.A.;
RT "Non-identity-mediated CRISPR-bacteriophage interaction mediated via the
RT Csy and Cas3 proteins.";
RL J. Bacteriol. 193:3433-3445(2011).
RN [3]
RP NO ROLE IN PHAGE PROTECTION, AND DISRUPTION PHENOTYPE.
RC STRAIN=UCBPP-PA14;
RX PubMed=21081758; DOI=10.1099/mic.0.045732-0;
RA Cady K.C., White A.S., Hammond J.H., Abendroth M.D., Karthikeyan R.S.,
RA Lalitha P., Zegans M.E., O'Toole G.A.;
RT "Prevalence, conservation and functional analysis of Yersinia and
RT Escherichia CRISPR regions in clinical Pseudomonas aeruginosa isolates.";
RL Microbiology 157:430-437(2011).
RN [4]
RP SUBUNIT, RNA-BINDING, AND MASS SPECTROMETRY.
RC STRAIN=UCBPP-PA14;
RX PubMed=21536913; DOI=10.1073/pnas.1102716108;
RA Wiedenheft B., van Duijn E., Bultema J.B., Waghmare S.P., Zhou K.,
RA Barendregt A., Westphal W., Heck A.J., Boekema E.J., Dickman M.J.,
RA Doudna J.A.;
RT "RNA-guided complex from a bacterial immune system enhances target
RT recognition through seed sequence interactions.";
RL Proc. Natl. Acad. Sci. U.S.A. 108:10092-10097(2011).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS), FUNCTION AS AN ENDONUCLEASE,
RP COFACTOR, RNA-BINDING, AND MUTAGENESIS OF HIS-29; ARG-102; GLN-104; SER-148
RP AND PHE-155.
RC STRAIN=UCBPP-PA14;
RX PubMed=20829488; DOI=10.1126/science.1192272;
RA Haurwitz R.E., Jinek M., Wiedenheft B., Zhou K., Doudna J.A.;
RT "Sequence- and structure-specific RNA processing by a CRISPR
RT endonuclease.";
RL Science 329:1355-1358(2010).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEX WITH RNA, FUNCTION IN
RP CRRNA FORMATION, SUBUNIT, RNA-BINDING, AND MUTAGENESIS OF HIS-29; SER-148;
RP SER-150; THR-151 AND TYR-176.
RC STRAIN=UCBPP-PA14;
RX PubMed=22522703; DOI=10.1038/emboj.2012.107;
RA Haurwitz R.E., Sternberg S.H., Doudna J.A.;
RT "Csy4 relies on an unusual catalytic dyad to position and cleave CRISPR
RT RNA.";
RL EMBO J. 31:2824-2832(2012).
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).
CC Processes pre-crRNA into individual crRNA units. Absolutely required
CC for crRNA production or stability. Upon expression in E.coli
CC endonucleolytically processes pre-crRNA, although disruption and
CC reconstitution experiments indicate that in situ other genes are also
CC required for processing. Yields 5'-hydroxy and 3'-phosphate groups. The
CC Csy ribonucleoprotein complex binds target ssDNA with high affinity but
CC target dsDNA with much lower affinity. {ECO:0000269|PubMed:20829488,
CC ECO:0000269|PubMed:21398535, ECO:0000269|PubMed:22522703}.
CC -!- COFACTOR:
CC Note=Metal-ion independent. Binds processed crRNA.
CC {ECO:0000269|PubMed:20829488};
CC -!- SUBUNIT: Part of the Csy ribonucleoprotein complex with a probable
CC stoichiometry of Csy1(1),Csy2(1),Csy3(6),Cas6/Csy4(1)-crRNA(1). A
CC Csy3(6),Cas6/Csy4(1)-crRNA(1) subcomplex is also formed.
CC {ECO:0000269|PubMed:21536913, ECO:0000269|PubMed:22522703}.
CC -!- INTERACTION:
CC Q02MM2; Q02MM1: csy3; NbExp=12; IntAct=EBI-15924852, EBI-15924841;
CC -!- MASS SPECTROMETRY: Mass=21533.8; Mass_error=1.9; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:21536913};
CC -!- DISRUPTION PHENOTYPE: Infection with phage DMS3 inhibits biofilm
CC formation; disrupting this gene restores biofilm formation despite DMS3
CC infection. Normal biofilm formation in the absence of phage infection.
CC Loss of production of crRNA in the presence or absence of phage.
CC Disruption of the entire Y.pestis-subtype CRISPR region disrupts crRNA
CC production but does not alter phage resistance (possibly OLNs
CC PA14_33350 to PA14_33310, plus the flanking CRISPR loci), indicating
CC this CRISPR is not involved in phage resistance.
CC {ECO:0000269|PubMed:21081758, ECO:0000269|PubMed:21398535}.
CC -!- MISCELLANEOUS: In this bacteria, Y.pestis-subtype CRISPRs do not confer
CC resistance to phage DMS3 or MP22, but instead are required for DMS3-
CC dependent inhibition of biofilm formation and possibly motility.
CC -!- SIMILARITY: Belongs to the CRISPR-associated endoribonuclease Cas6
CC family. Cas6f/Csy4, subtype I-F/Ypest subfamily. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=ABJ11605.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305};
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DR EMBL; CP000438; ABJ11605.1; ALT_INIT; Genomic_DNA.
DR RefSeq; WP_016254196.1; NZ_CP034244.1.
DR PDB; 2XLI; X-ray; 2.33 A; A=1-187.
DR PDB; 2XLJ; X-ray; 2.60 A; A=1-187.
DR PDB; 2XLK; X-ray; 1.80 A; A/B=1-187.
DR PDB; 4AL5; X-ray; 2.00 A; A=1-187.
DR PDB; 4AL6; X-ray; 2.63 A; A=1-187.
DR PDB; 4AL7; X-ray; 2.32 A; A=1-187.
DR PDB; 5UZ9; EM; 3.40 A; L=1-187.
DR PDB; 6B44; EM; 2.90 A; L=1-187.
DR PDB; 6B45; EM; 3.50 A; L=1-187.
DR PDB; 6B46; EM; 3.10 A; L=1-187.
DR PDB; 6B47; EM; 3.20 A; L=1-187.
DR PDB; 6B48; EM; 3.60 A; L=1-187.
DR PDB; 6NE0; EM; 3.40 A; L=1-187.
DR PDBsum; 2XLI; -.
DR PDBsum; 2XLJ; -.
DR PDBsum; 2XLK; -.
DR PDBsum; 4AL5; -.
DR PDBsum; 4AL6; -.
DR PDBsum; 4AL7; -.
DR PDBsum; 5UZ9; -.
DR PDBsum; 6B44; -.
DR PDBsum; 6B45; -.
DR PDBsum; 6B46; -.
DR PDBsum; 6B47; -.
DR PDBsum; 6B48; -.
DR PDBsum; 6NE0; -.
DR AlphaFoldDB; Q02MM2; -.
DR SMR; Q02MM2; -.
DR DIP; DIP-59681N; -.
DR IntAct; Q02MM2; 6.
DR PRIDE; Q02MM2; -.
DR EnsemblBacteria; ABJ11605; ABJ11605; PA14_33300.
DR KEGG; pau:PA14_33300; -.
DR HOGENOM; CLU_108958_0_0_6; -.
DR BioCyc; PAER208963:G1G74-2802-MON; -.
DR Proteomes; UP000000653; Chromosome.
DR GO; GO:0004519; F:endonuclease activity; IEA:UniProtKB-KW.
DR GO; GO:0003723; F:RNA binding; IEA:UniProtKB-KW.
DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-KW.
DR GO; GO:0043571; P:maintenance of CRISPR repeat elements; IEA:InterPro.
DR CDD; cd09739; Cas6_I-F; 1.
DR Gene3D; 3.30.70.2540; -; 1.
DR InterPro; IPR013396; CRISPR-assoc_prot_Csy4.
DR InterPro; IPR042564; CRISPR-Cas6/Csy4_sf.
DR Pfam; PF09618; Cas_Csy4; 1.
DR TIGRFAMs; TIGR02563; cas_Csy4; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antiviral defense; Endonuclease; Hydrolase; Nuclease;
KW RNA-binding.
FT CHAIN 1..187
FT /note="CRISPR-associated endonuclease Cas6/Csy4"
FT /id="PRO_0000417882"
FT ACT_SITE 29
FT /note="Proton acceptor"
FT /evidence="ECO:0000305"
FT SITE 148
FT /note="Substrate binding"
FT MUTAGEN 29
FT /note="H->A: No pre-crRNA cleavage, still binds crRNA. Does
FT not support formation of the Csy ribonucleoprotein
FT complex."
FT /evidence="ECO:0000269|PubMed:20829488,
FT ECO:0000269|PubMed:21398535, ECO:0000269|PubMed:22522703"
FT MUTAGEN 29
FT /note="H->D: Cleaves pre-crRNA 910-fold slower."
FT /evidence="ECO:0000269|PubMed:20829488,
FT ECO:0000269|PubMed:21398535, ECO:0000269|PubMed:22522703"
FT MUTAGEN 29
FT /note="H->K: Cleaves pre-crRNA 130-fold slower."
FT /evidence="ECO:0000269|PubMed:20829488,
FT ECO:0000269|PubMed:21398535, ECO:0000269|PubMed:22522703"
FT MUTAGEN 49
FT /note="E->A: No biofilm formation upon phage infection, no
FT crRNA formed."
FT MUTAGEN 49
FT /note="E->K: Restores biofilm formation upon phage
FT infection, crRNA forms."
FT MUTAGEN 102
FT /note="R->A: Loss of pre-crRNA cleavage, still binds
FT crRNA."
FT /evidence="ECO:0000269|PubMed:20829488"
FT MUTAGEN 104
FT /note="Q->A: No loss of pre-crRNA cleavage, still binds
FT crRNA."
FT /evidence="ECO:0000269|PubMed:20829488"
FT MUTAGEN 148
FT /note="S->A: Cleaves pre-crRNA 8300-fold slower."
FT /evidence="ECO:0000269|PubMed:20829488,
FT ECO:0000269|PubMed:22522703"
FT MUTAGEN 148
FT /note="S->C: No pre-crRNA cleavage, still binds crRNA."
FT /evidence="ECO:0000269|PubMed:20829488,
FT ECO:0000269|PubMed:22522703"
FT MUTAGEN 150
FT /note="S->A: Cleaves pre-crRNA 350-fold slower."
FT /evidence="ECO:0000269|PubMed:22522703"
FT MUTAGEN 151
FT /note="T->A: Cleaves pre-crRNA 380-fold slower."
FT /evidence="ECO:0000269|PubMed:22522703"
FT MUTAGEN 155
FT /note="F->A: Very little pre-crRNA cleavage, still binds
FT crRNA."
FT /evidence="ECO:0000269|PubMed:20829488"
FT MUTAGEN 176
FT /note="Y->A: Cleaves pre-crRNA 130-fold slower."
FT /evidence="ECO:0000269|PubMed:22522703"
FT MUTAGEN 176
FT /note="Y->F: Cleaves pre-crRNA 13-fold slower."
FT /evidence="ECO:0000269|PubMed:22522703"
FT STRAND 3..9
FT /evidence="ECO:0007829|PDB:2XLK"
FT STRAND 13..15
FT /evidence="ECO:0007829|PDB:2XLK"
FT HELIX 17..35
FT /evidence="ECO:0007829|PDB:2XLK"
FT STRAND 38..40
FT /evidence="ECO:0007829|PDB:6B45"
FT STRAND 41..43
FT /evidence="ECO:0007829|PDB:2XLK"
FT TURN 49..52
FT /evidence="ECO:0007829|PDB:2XLK"
FT STRAND 56..62
FT /evidence="ECO:0007829|PDB:2XLK"
FT HELIX 64..71
FT /evidence="ECO:0007829|PDB:2XLK"
FT HELIX 74..76
FT /evidence="ECO:0007829|PDB:4AL5"
FT HELIX 77..82
FT /evidence="ECO:0007829|PDB:2XLK"
FT STRAND 83..85
FT /evidence="ECO:0007829|PDB:4AL5"
FT STRAND 93..95
FT /evidence="ECO:0007829|PDB:6B47"
FT STRAND 100..103
FT /evidence="ECO:0007829|PDB:2XLK"
FT STRAND 105..107
FT /evidence="ECO:0007829|PDB:6B47"
FT HELIX 109..120
FT /evidence="ECO:0007829|PDB:2XLK"
FT HELIX 124..130
FT /evidence="ECO:0007829|PDB:2XLK"
FT HELIX 133..135
FT /evidence="ECO:0007829|PDB:4AL5"
FT STRAND 143..147
FT /evidence="ECO:0007829|PDB:2XLK"
FT TURN 149..151
FT /evidence="ECO:0007829|PDB:2XLK"
FT STRAND 154..161
FT /evidence="ECO:0007829|PDB:2XLK"
FT STRAND 165..167
FT /evidence="ECO:0007829|PDB:2XLK"
FT STRAND 174..176
FT /evidence="ECO:0007829|PDB:6NE0"
FT STRAND 179..181
FT /evidence="ECO:0007829|PDB:2XLK"
SQ SEQUENCE 187 AA; 21391 MW; 0C4A370883007481 CRC64;
MDHYLDIRLR PDPEFPPAQL MSVLFGKLHQ ALVAQGGDRI GVSFPDLDES RSRLGERLRI
HASADDLRAL LARPWLEGLR DHLQFGEPAV VPHPTPYRQV SRVQAKSNPE RLRRRLMRRH
DLSEEEARKR IPDTVARALD LPFVTLRSQS TGQHFRLFIR HGPLQVTAEE GGFTCYGLSK
GGFVPWF
