RT67_ECOLX
ID RT67_ECOLX Reviewed; 586 AA.
AC P21325;
DT 01-MAY-1991, integrated into UniProtKB/Swiss-Prot.
DT 03-AUG-2022, sequence version 2.
DT 03-AUG-2022, entry version 83.
DE RecName: Full=Retron Ec67 protein {ECO:0000303|PubMed:2466332};
DE AltName: Full=ORF4-Ec67 RT {ECO:0000303|PubMed:1701261};
DE Includes:
DE RecName: Full=RNA-directed DNA polymerase;
DE EC=2.7.7.49 {ECO:0000255|PROSITE-ProRule:PRU00405, ECO:0000269|PubMed:1378431, ECO:0000305|PubMed:2466332};
DE AltName: Full=Reverse transcriptase {ECO:0000303|PubMed:2466332};
DE Short=RT {ECO:0000303|PubMed:2466332};
DE Includes:
DE RecName: Full=Ribonuclease H;
DE Short=RNase H;
DE EC=3.1.26.4 {ECO:0000305|PubMed:2466332};
GN Name=ret {ECO:0000303|PubMed:1722556};
GN ORFNames=Ga0175966_113075, RG66_23265 {ECO:0000303|Ref.4};
OS Escherichia coli.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=562;
RN [1] {ECO:0000312|EMBL:AAA56874.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROBABLE FUNCTION AS A REVERSE
RP TRANSCRIPTASE, PROBABLE CATALYTIC ACTIVITY, DOMAIN, DISRUPTION PHENOTYPE,
RP AND MUTAGENESIS OF 430-ALA--SER-586.
RC STRAIN=O1:NM / CL-1;
RX PubMed=2466332; DOI=10.1126/science.2466332;
RA Lampson B.C., Sun J., Hsu M.-Y., Vallejo-Ramirez J., Inouye S., Inouye M.;
RT "Reverse transcriptase in a clinical strain of Escherichia coli: production
RT of branched RNA-linked msDNA.";
RL Science 243:1033-1038(1989).
RN [2] {ECO:0000312|EMBL:AAA23403.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=O1:NM / CL-1;
RX PubMed=1701261; DOI=10.1073/pnas.87.23.9454;
RA Hsu M.-Y., Inouye M., Inouye S.;
RT "Retron for the 67-base multicopy single-stranded DNA from Escherichia
RT coli: a potential transposable element encoding both reverse transcriptase
RT and Dam methylase functions.";
RL Proc. Natl. Acad. Sci. U.S.A. 87:9454-9458(1990).
RN [3] {ECO:0000312|EMBL:X60207}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=R:H2 / Isolate 317 Brazil;
RX PubMed=1722556; DOI=10.1111/j.1365-2958.1991.tb00810.x;
RA Lim D.;
RT "Structure of two retrons of Escherichia coli and their common chromosomal
RT insertion site.";
RL Mol. Microbiol. 5:1863-1872(1991).
RN [4] {ECO:0000312|EMBL:APL54601.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=S10;
RA Jiang J., Xu Z., Zhou Y., Zhao H., Leung F.C.;
RT "Vertical and Horizontal Evolutionary Story implied from E. coli complete
RT genomes.";
RL Submitted (NOV-2014) to the EMBL/GenBank/DDBJ databases.
RN [5]
RP FUNCTION, AND MSDNA-BINDING.
RC STRAIN=O1:NM / CL-1;
RX PubMed=1692831; DOI=10.1016/s0021-9258(19)38915-x;
RA Lampson B.C., Viswanathan M., Inouye M., Inouye S.;
RT "Reverse transcriptase from Escherichia coli exists as a complex with msDNA
RT and is able to synthesize double-stranded DNA.";
RL J. Biol. Chem. 265:8490-8496(1990).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, AND DISRUPTION PHENOTYPE.
RC STRAIN=O1:NM / CL-1;
RX PubMed=1378431; DOI=10.1016/s0021-9258(19)49642-7;
RA Hsu M.Y., Eagle S.G., Inouye M., Inouye S.;
RT "Cell-free synthesis of the branched RNA-linked msDNA from retron Ec67 of
RT Escherichia coli.";
RL J. Biol. Chem. 267:13823-13829(1992).
RN [7]
RP FUNCTION IN ANTIVIRAL DEFENSE, AND DOMAIN.
RC STRAIN=S10;
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: Reverse transcriptase (RT) component of antiviral defense
CC system retron Ec67, minimally composed of a non-coding RNA (ncRNA) and
CC this RT. Expression of these 2 elements confers protection against
CC bacteriophage T5. At multiplicity of infection (MOI) of 0.02 cultures
CC grow normally when infected with T5 without collapsing, at MOI 2
CC cultures enter growth stasis (PubMed:33157039). Responsible for
CC synthesis of msDNA-Ec67 (a branched molecule with RNA linked by a
CC 2',5'-phosphodiester bond to ssDNA). The retron transcript serves as
CC primer (from a conserved internal G residue) and template for the
CC reaction, and codes for the RT (PubMed:2466332, PubMed:1378431,
CC PubMed:1692831). Can use other retrons as substrate (msDNA-Mx162 and
CC msDNA-Ec86). Also able to synthesize DNA from a DNA template at least
CC in vitro, although the enzyme is less active with a DNA template
CC (PubMed:1692831). {ECO:0000269|PubMed:1378431,
CC ECO:0000269|PubMed:1692831, ECO:0000269|PubMed:2466332,
CC ECO:0000269|PubMed:33157039}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 2'-deoxyribonucleoside 5'-triphosphate + DNA(n) =
CC diphosphate + DNA(n+1); Xref=Rhea:RHEA:22508, Rhea:RHEA-COMP:17339,
CC Rhea:RHEA-COMP:17340, ChEBI:CHEBI:33019, ChEBI:CHEBI:61560,
CC ChEBI:CHEBI:173112; EC=2.7.7.49; Evidence={ECO:0000255|PROSITE-
CC ProRule:PRU00405, ECO:0000269|PubMed:1378431,
CC ECO:0000305|PubMed:2466332};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Endonucleolytic cleavage to 5'-phosphomonoester.; EC=3.1.26.4;
CC Evidence={ECO:0000305|PubMed:2466332};
CC -!- DOMAIN: The N-terminus is the reverse transcriptase, the C-terminus
CC (downstream of residue 430) is also required for msDNA synthesis
CC (PubMed:2466332). The C-terminus was originally suggested to have RNase
CC H activity, but may have another nuclease activity instead (Probable).
CC {ECO:0000269|PubMed:2466332, ECO:0000305|PubMed:2466332,
CC ECO:0000305|PubMed:33157039}.
CC -!- DISRUPTION PHENOTYPE: No synthesis of msDNA.
CC {ECO:0000269|PubMed:1378431, ECO:0000269|PubMed:2466332}.
CC -!- MISCELLANEOUS: Retrons may be the ancestors of retrovirus.
CC -!- SIMILARITY: Belongs to the bacterial reverse transcriptase family.
CC {ECO:0000305}.
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DR EMBL; M24363; AAA56874.1; -; Genomic_DNA.
DR EMBL; M55249; AAA23403.1; -; Genomic_DNA.
DR EMBL; X60207; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR EMBL; CP010229; APL54601.1; -; Genomic_DNA.
DR PIR; S16654; S16654.
DR RefSeq; WP_016235542.1; NZ_UNPP01000023.1.
DR AlphaFoldDB; P21325; -.
DR GO; GO:0003723; F:RNA binding; IEA:InterPro.
DR GO; GO:0003964; F:RNA-directed DNA polymerase activity; IEA:UniProtKB-KW.
DR GO; GO:0004523; F:RNA-DNA hybrid ribonuclease activity; IEA:UniProtKB-EC.
DR CDD; cd03487; RT_Bac_retron_II; 1.
DR InterPro; IPR043502; DNA/RNA_pol_sf.
DR InterPro; IPR000123; Reverse_transcriptase_msDNA.
DR InterPro; IPR000477; RT_dom.
DR Pfam; PF00078; RVT_1; 1.
DR PRINTS; PR00866; RNADNAPOLMS.
DR SUPFAM; SSF56672; SSF56672; 1.
DR PROSITE; PS50878; RT_POL; 1.
PE 1: Evidence at protein level;
KW Antiviral defense; Endonuclease; Hydrolase; Magnesium; Metal-binding;
KW Multifunctional enzyme; Nuclease; Nucleotidyltransferase; RNA-binding;
KW RNA-directed DNA polymerase; Transferase; Transposable element.
FT CHAIN 1..586
FT /note="Retron Ec67 protein"
FT /id="PRO_0000097508"
FT DOMAIN 29..262
FT /note="Reverse transcriptase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00405"
FT BINDING 120
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00405"
FT BINDING 201
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00405"
FT BINDING 202
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00405"
FT MUTAGEN 430..586
FT /note="Missing: No synthesis of msDNA."
FT /evidence="ECO:0000269|PubMed:2466332"
FT CONFLICT 25
FT /note="V -> I (in Ref. 1; AAA56874, 2; AAA23403 and 3;
FT X60207)"
FT CONFLICT 47
FT /note="T -> R (in Ref. 1; AAA56874)"
FT /evidence="ECO:0000305"
FT CONFLICT 152
FT /note="T -> R (in Ref. 1; AAA56874)"
FT /evidence="ECO:0000305"
FT CONFLICT 211
FT /note="T -> R (in Ref. 1; AAA56874)"
FT /evidence="ECO:0000305"
FT CONFLICT 261
FT /note="T -> R (in Ref. 1; AAA56874)"
FT /evidence="ECO:0000305"
FT CONFLICT 341
FT /note="H -> D (in Ref. 1; AAA56874)"
FT /evidence="ECO:0000305"
FT CONFLICT 373
FT /note="T -> R (in Ref. 1; AAA56874)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 586 AA; 67220 MW; 9EA0F64DE98CE383 CRC64;
MTKTSKLDAL RAATSREDLA KILDVKLVFL TNVLYRIGSD NQYTQFTIPK KGKGVRTISA
PTDRLKDIQR RICDLLSDCR DEIFAIRKIS NNYSFGFERG KSIILNAYKH RGKQIILNID
LKDFFESFNF GRVRGYFLSN QDFLLNPVVA TTLAKAACYN GTLPQGSPCS PIISNLICNI
MDMRLAKLAK KYGCTYSRYA DDITISTNKN TFPLEMATVQ PEGVVLGKVL VKEIENSGFE
INDSKTRLTY KTSRQEVTGL TVNRIVNIDR CYYKKTRALA HALYRTGEYK VPDENGVLVS
GGLDKLEGMF GFIDQVDKFN NIKKKLNKQP DRYVLTNATL HGFKLKLNAR EKAYSKFIYY
KFFHGNTCPT IITEGKTDRI YLKAALHSLE TSYPELFREK TDSKKKEINL NIFKSNEKTK
YFLDLSGGTA DLKKFVERYK NNYASYYGSV PKQPVIMVLD NDTGPSDLLN FLRNKVKSCP
DDVTEMRKMK YIHVFYNLYI VLTPLSPSGE QTSMEDLFPK DILDIKIDGK KFNKNNDGDS
KTEYGKHIFS MRVVRDKKRK IDFKAFCCIF DAIKDIKEHY KLMLNS
