OLD8_ECOLX
ID OLD8_ECOLX Reviewed; 750 AA.
AC P0DV58;
DT 03-AUG-2022, integrated into UniProtKB/Swiss-Prot.
DT 03-AUG-2022, sequence version 1.
DT 03-AUG-2022, entry version 1.
DE RecName: Full=Retron Eco8 OLD nuclease {ECO:0000303|PubMed:33157039};
DE EC=3.1.-.- {ECO:0000305};
GN Name=old {ECO:0000303|PubMed:33157039};
GN ORFNames=ERS139198_01420 {ECO:0000303|Ref.1}, Ga0119705_103344;
OS Escherichia coli.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=562;
RN [1] {ECO:0000312|EMBL:CUA03350.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=200499;
RA Babu N.S., Beckwith C.J., Beseler K.G., Brison A., Carone J.V.,
RA Caskin T.P., Diamond M., Durham M.E., Foxe J.M., Go M., Henderson B.A.,
RA Jones I.B., McGettigan J.A., Micheletti S.J., Nasrallah M.E., Ortiz D.,
RA Piller C.R., Privatt S.R., Schneider S.L., Sharp S., Smith T.C.,
RA Stanton J.D., Ullery H.E., Wilson R.J., Serrano M.G., Buck G., Lee V.,
RA Wang Y., Carvalho R., Voegtly L., Shi R., Duckworth R., Johnson A.,
RA Loviza R., Walstead R., Shah Z., Kiflezghi M., Wade K., Ball S.L.,
RA Bradley K.W., Asai D.J., Bowman C.A., Russell D.A., Pope W.H.,
RA Jacobs-Sera D., Hendrix R.W., Hatfull G.F.;
RL Submitted (AUG-2015) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION IN ANTIVIRAL DEFENSE, IDENTIFICATION AS A RETRON, AND MUTAGENESIS
RP OF LYS-36.
RC STRAIN=200499;
RX PubMed=33157039; DOI=10.1016/j.cell.2020.09.065;
RA Millman A., Bernheim A., Stokar-Avihail A., Fedorenko T., Voichek M.,
RA Leavitt A., Oppenheimer-Shaanan Y., Sorek R.;
RT "Bacterial Retrons Function In Anti-Phage Defense.";
RL Cell 183:1551-1561(2020).
CC -!- FUNCTION: Probable nuclease member of antiviral defense system retron
CC Eco8, composed of an reverse transcriptase (RT), this nuclease and a
CC non-coding RNA (ncRNA) encoded between them. Expression of retron Eco8
CC confers protection against bacteriophages T4, T6, T7 and SECphi4,
CC SECphi6 and SECphi18. At multiplicity of infection (MOI) of 0.02
CC cultures slow growth when infected with SECphi4 but do not collapse, at
CC MOI 2 cultures collapse. When the retron is cloned in another E.coli
CC strain synthesizes msDNA (a branched RNA linked by a 2',5'-
CC phosphodiester bond to a single-stranded DNA). The retron transcript
CC serves as primer and template to the reaction, and codes for the RT.
CC {ECO:0000269|PubMed:33157039}.
CC -!- COFACTOR:
CC Name=a divalent metal cation; Xref=ChEBI:CHEBI:60240;
CC Evidence={ECO:0000250|UniProtKB:E8PLM2};
CC Note=Probably binds 2 metal cations. {ECO:0000250|UniProtKB:E8PLM2};
CC -!- SUBUNIT: Homodimer. {ECO:0000250|UniProtKB:E8PLM2}.
CC -!- SIMILARITY: Belongs to the class 1 OLD nuclease family.
CC {ECO:0000305|PubMed:33157039}.
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DR EMBL; CYGJ01000003; CUA03350.1; -; Genomic_DNA.
PE 1: Evidence at protein level;
KW Antiviral defense; ATP-binding; Hydrolase; Metal-binding; Nuclease;
KW Nucleotide-binding.
FT CHAIN 1..750
FT /note="Retron Eco8 OLD nuclease"
FT /id="PRO_0000456030"
FT REGION 1..173
FT /note="ATPase domain N-terminus"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT REGION 174..260
FT /note="Dimerization domain"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT REGION 261..390
FT /note="ATPase domain C-terminus"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT REGION 391..704
FT /note="Toprim domain"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT BINDING 33..37
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT BINDING 398
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT BINDING 402
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT BINDING 450
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT BINDING 452
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT BINDING 623
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT BINDING 641
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:E8PLM2"
FT MUTAGEN 36
FT /note="K->A: No longer protects against SECphi6 infection."
FT /evidence="ECO:0000269|PubMed:33157039"
SQ SEQUENCE 750 AA; 87256 MW; FA1B2278E666C066 CRC64;
MTIESIRVKN LLSFDDVILR DFRDINCIIG RNNVGKSNLL KVIRYFYAKL ENKKVIPLDF
HTNYNAVGEI TFTFDTTRIK KIVTSRKNNG RFHKHIYNTL FKSSSVKLNF EELIARKNST
NKSFFSLTLT ICKDDSVMWS VDDPKVRSLL ATLYPFLYIE TRHIDLYDWN PIWKLISNLN
SFNFDDVDHD ELVNFLDEKI SSRKGDYKKY IDRVVSVIDT KPYTYKEKVI NYIKVAIKGD
SFVNAGEELF TQSDGTNSNK FLETLLHLLI TLTRTEFISP IVYIDEPEVG LHPKLAESFV
SNLNKIYSKF KKTSELSGPG RYKTPYPNIF YSTHSPSILK QTIKLFGKDQ QVLHFSKKKD
GSTRVNKINS TYSDERFLNI FSDNEARLFF SEYIVFVEGA TELELFRNLS LLNLYPAFSL
ADIYDANEVI LANINPGYSK ASIPFVIIKD IDTLIDYSIK TEKFSLRPLF EKMIKELTKE
FDYYDTGFGR VRKEIDLFSD IQSSTKKHMD SGLFFKRFSL HNLSSRINKV SRKLNRYFMT
TTIEGALINE QSLPYFFNWI GDVILTQMTI NNPNPDKFIE AMRRRYNIKS QVVPLFKSVF
CIGLNHPVYS SAVDKQALRI KLSFLNYLKR KVYSDFNNEK EIVLALRLAF GGKTETQYTL
DKLRKDGEAE LFREKIKNYK NNELFFLEPQ MTKTSGWVTT FLNYTIEKIT SEESDDDRIR
QKLSFIFPEI ISIIEQASSS IEAEESSLTG
