PYCC1_ECOLX
ID PYCC1_ECOLX Reviewed; 450 AA.
AC P0DV24;
DT 25-MAY-2022, integrated into UniProtKB/Swiss-Prot.
DT 25-MAY-2022, sequence version 1.
DT 03-AUG-2022, entry version 2.
DE RecName: Full=Cytidylate cyclase {ECO:0000303|PubMed:34644530};
DE EC=4.6.1.6 {ECO:0000269|PubMed:34644530};
DE AltName: Full=Cyclic CMP synthase;
DE Short=cCMP synthase {ECO:0000303|PubMed:34644530};
DE AltName: Full=EcPycC {ECO:0000303|PubMed:34644530};
GN Name=pycC {ECO:0000303|PubMed:34644530}; ORFNames=Ga0132381_1285;
OS Escherichia coli.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=562;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=E831;
RA Hur Y.J.;
RL Submitted (AUG-2015) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, COFACTOR, ACTIVITY
RP REGULATION BY VIRAL INFECTION, BIOPHYSICOCHEMICAL PROPERTIES, ANTIVIRAL
RP DEFENSE, DOMAIN, CLASSIFICATION, AND MUTAGENESIS OF PHE-100; ASP-102;
RP ARG-142; MET-144; ASP-146; ASP-230 AND GLN-237.
RC STRAIN=E831;
RX PubMed=34644530; DOI=10.1016/j.cell.2021.09.031;
RA Tal N., Morehouse B.R., Millman A., Stokar-Avihail A., Avraham C.,
RA Fedorenko T., Yirmiya E., Herbst E., Brandis A., Mehlman T.,
RA Oppenheimer-Shaanan Y., Keszei A.F.A., Shao S., Amitai G., Kranzusch P.J.,
RA Sorek R.;
RT "Cyclic CMP and cyclic UMP mediate bacterial immunity against phages.";
RL Cell 0:0-0(2021).
CC -!- FUNCTION: Pycsar (pyrimidine cyclase system for antiphage resistance)
CC provides immunity against bacteriophage. The pyrimidine cyclase (PycC)
CC synthesizes cyclic nucleotides in response to infection; these serve as
CC specific second messenger signals. The signal activates the adjacent
CC effector, leading to bacterial cell death and abortive phage infection.
CC A clade E Pycsar system. {ECO:0000269|PubMed:34644530}.
CC -!- FUNCTION: The pyrimidine cyclase gene of a two-gene Pycsar system,
CC generates cyclic CMP (cCMP) from CTP in response to bacteriophage
CC infection. Has little to no activity on ATP, GTP or UTP. Expression of
CC this and adjacent effector EcPycTM (AC P0DV25) confers resistance to
CC bacteriophage P1 and T5; expression of this gene alone does not confer
CC resistance. When cells expressing the Pycsar system are infected by
CC phage T5 at low multiplicity of infection (0.2 MOI) the culture
CC survives, at 2.0 MOI bacteria enter growth arrest. The same cells enter
CC growth arrest after exposure to 250 uM cCMP but not cUMP; thus the
CC effector protein responds only to the cNMP produced by its cognate NTP
CC cyclase. Some of the cells treated with cCMP have abnormal membrane
CC protrusions. {ECO:0000269|PubMed:34644530}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=CTP = 3',5'-cyclic CMP + diphosphate; Xref=Rhea:RHEA:14737,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:37563, ChEBI:CHEBI:58003; EC=4.6.1.6;
CC Evidence={ECO:0000269|PubMed:34644530};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:34644530};
CC Note=Cannot be replaced by Mg(2+). {ECO:0000269|PubMed:34644530};
CC -!- ACTIVITY REGULATION: In E.coli strain MG1655 transformed with both
CC genes cCMP appears between 15 and 30 minutes after infection with phage
CC T5 (at protein level). No cCMP accumulates in uninfected cells.
CC {ECO:0000269|PubMed:34644530}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 7.5-9.0. {ECO:0000269|PubMed:34644530};
CC -!- SUBUNIT: Homodimer. {ECO:0000250|UniProtKB:A0A0J5ZXG5}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- DOMAIN: Has an N-terminal nucleotide cyclase domain and a C-terminal
CC nucleotide sensor domain (AGS-C); removal of the latter leads to loss
CC of phage P1 and T5 resistance. {ECO:0000269|PubMed:34644530}.
CC -!- MISCELLANEOUS: T5 phage that escape Pycsar have mutated major capsid
CC protein pb8 (D20, AC Q6QGD8); infection with these phage elicits less
CC cCMP production. Ectopic expression of the capsid protein in addition
CC to both Pycsar genes does not induce effector-mediated toxicity,
CC suggesting another factor is necessary. {ECO:0000269|PubMed:34644530}.
CC -!- SIMILARITY: Belongs to the adenylyl cyclase class-4/guanylyl cyclase
CC family. Pyrimidine cyclase subfamily. {ECO:0000305|PubMed:34644530}.
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DR EMBL; CXXA01000028; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0016829; F:lyase activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0051607; P:defense response to virus; IEA:UniProtKB-KW.
DR Gene3D; 3.30.70.1230; -; 1.
DR InterPro; IPR001054; A/G_cyclase.
DR InterPro; IPR040511; AGS_C.
DR InterPro; IPR029787; Nucleotide_cyclase.
DR Pfam; PF18134; AGS_C; 1.
DR Pfam; PF00211; Guanylate_cyc; 1.
DR SUPFAM; SSF55073; SSF55073; 1.
DR PROSITE; PS50125; GUANYLATE_CYCLASE_2; 1.
PE 1: Evidence at protein level;
KW Antiviral defense; Cytoplasm; Lyase; Manganese; Metal-binding;
KW Nucleotide-binding.
FT CHAIN 1..450
FT /note="Cytidylate cyclase"
FT /id="PRO_0000455218"
FT DOMAIN 97..236
FT /note="Guanylate cyclase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00099"
FT REGION 318..450
FT /note="AGS-C domain"
FT /evidence="ECO:0000305|PubMed:34644530"
FT BINDING 100
FT /ligand="a ribonucleoside 5'-triphosphate"
FT /ligand_id="ChEBI:CHEBI:61557"
FT /evidence="ECO:0000250|UniProtKB:P0DV40,
FT ECO:0000305|PubMed:34644530"
FT BINDING 102
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000305|PubMed:34644530"
FT BINDING 102
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000305|PubMed:34644530"
FT BINDING 103
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000305|PubMed:34644530"
FT BINDING 146
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="1"
FT /evidence="ECO:0000305|PubMed:34644530"
FT BINDING 146
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /ligand_label="2"
FT /evidence="ECO:0000305|PubMed:34644530"
FT MUTAGEN 100
FT /note="F->A: Loss of phage T5 resistance, wild-type cCMP
FT production."
FT /evidence="ECO:0000269|PubMed:34644530"
FT MUTAGEN 102
FT /note="D->A: Loss of phage T5 resistance, no cCMP
FT production."
FT /evidence="ECO:0000269|PubMed:34644530"
FT MUTAGEN 142
FT /note="R->A: Loss of phage T5 resistance, no cCMP
FT production."
FT /evidence="ECO:0000269|PubMed:34644530"
FT MUTAGEN 144
FT /note="M->A: Loss of phage T5 resistance, wild-type cCMP
FT production."
FT /evidence="ECO:0000269|PubMed:34644530"
FT MUTAGEN 146
FT /note="D->A: Loss of phage T5 resistance, no cCMP
FT production."
FT /evidence="ECO:0000269|PubMed:34644530"
FT MUTAGEN 230
FT /note="D->A: Wild-type cCMP production."
FT /evidence="ECO:0000269|PubMed:34644530"
FT MUTAGEN 237
FT /note="Q->A: Wild-type cCMP production."
FT /evidence="ECO:0000269|PubMed:34644530"
SQ SEQUENCE 450 AA; 50981 MW; 1697152B15D02D3D CRC64;
MSFKDVTAKN FKGLKNVSLK KSMAMEGHTL VGTEARLGDA FELCESFSTS PSNIIEYEYQ
EEIRPFFQKA GLNKHSIGTH PELTGLGVGM IYNQYTVTMF VDIRKSSRLS LLLPLEQVYV
VKNRILQACI DIVRALDGYP HRLMGDALMA FFGRSDVSKE DAIADAINAA STLRLILMDY
IFPSLNEDIG EQIDLGVRIG LDYGAEDEVV WGNFGLGSFC EVTALGLPVD MTAKLQQLAD
KNTAMLGQGI LDYIDFPEEY TKPKVKSGEE LKYIIPNITN KEGQPINRRI RLLNMARYQE
LLPFKLNDKK MASAILYPNQ FNFECFVIED NKEVLYNSVS RFLPKKRRLT FKLSIYPGPG
IGDLKIIFCK RNHGQEAKDD LSEDYSISIE DNKLIRVKNA DNLSLLRKDG CYVLTVPEET
LFRGLHTMEV IVRGNHETLF YRNIIGVYIK
