DPOL_THEFM
ID DPOL_THEFM Reviewed; 1523 AA.
AC P74918;
DT 15-DEC-1998, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1997, sequence version 1.
DT 25-MAY-2022, entry version 127.
DE RecName: Full=DNA polymerase;
DE EC=2.7.7.7 {ECO:0000305|Ref.1};
DE AltName: Full=Pol Tfu {ECO:0000303|PubMed:10644683};
DE Contains:
DE RecName: Full=Homing endonuclease PI-TfuI {ECO:0000303|PubMed:12654995};
DE EC=3.1.11.- {ECO:0000269|Ref.1};
DE AltName: Full=Tfu pol-1 intein {ECO:0000303|PubMed:10644683};
DE Contains:
DE RecName: Full=Homing endonuclease PI-TfuII {ECO:0000303|PubMed:12654995};
DE EC=3.1.11.- {ECO:0000269|Ref.1};
DE AltName: Full=Tfu pol-2 intein {ECO:0000303|PubMed:10644683};
GN Name=pol;
OS Thermococcus fumicolans.
OC Archaea; Euryarchaeota; Thermococci; Thermococcales; Thermococcaceae;
OC Thermococcus.
OX NCBI_TaxID=46540;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ST557;
RA Cambon M., Querellou J.;
RL Submitted (FEB-1996) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP CHARACTERIZATION OF INTEIN ENDONUCLEASE ACTIVITY, FUNCTION, CATALYTIC
RP ACTIVITY, COFACTOR, AND PROTEIN SPLICING.
RC STRAIN=ST557;
RX PubMed=10644683; DOI=10.1074/jbc.275.4.2335;
RA Saves I., Ozanne V., Dietrich J., Masson J.-M.;
RT "Inteins of Thermococcus fumicolans DNA polymerase are endonucleases with
RT distinct enzymatic behaviors.";
RL J. Biol. Chem. 275:2335-2341(2000).
RN [3]
RP NOMENCLATURE.
RX PubMed=12654995; DOI=10.1093/nar/gkg274;
RA Roberts R.J., Belfort M., Bestor T., Bhagwat A.S., Bickle T.A.,
RA Bitinaite J., Blumenthal R.M., Degtyarev S.K., Dryden D.T., Dybvig K.,
RA Firman K., Gromova E.S., Gumport R.I., Halford S.E., Hattman S.,
RA Heitman J., Hornby D.P., Janulaitis A., Jeltsch A., Josephsen J., Kiss A.,
RA Klaenhammer T.R., Kobayashi I., Kong H., Krueger D.H., Lacks S.,
RA Marinus M.G., Miyahara M., Morgan R.D., Murray N.E., Nagaraja V.,
RA Piekarowicz A., Pingoud A., Raleigh E., Rao D.N., Reich N., Repin V.E.,
RA Selker E.U., Shaw P.C., Stein D.C., Stoddard B.L., Szybalski W.,
RA Trautner T.A., Van Etten J.L., Vitor J.M., Wilson G.G., Xu S.Y.;
RT "A nomenclature for restriction enzymes, DNA methyltransferases, homing
RT endonucleases and their genes.";
RL Nucleic Acids Res. 31:1805-1812(2003).
CC -!- FUNCTION: Spliced protein is a DNA polymerase used in PCR reactions.
CC {ECO:0000305|Ref.1}.
CC -!- FUNCTION: PI-TfuI recognizes and cleaves a minimal sequence of 16 base
CC pairs (bp, 5'-TTTTAGGTCGCTATAT-3') cutting after T-8 in supercoiled DNA
CC with either Mn(2+) or Mg(2+) as cofactor. Yields 5'-phosphate and 3'-OH
CC ends. It only cleaves linear DNA with Mn(2+) and requires a 19-bp
CC minimal recognition sequence. {ECO:0000269|PubMed:10644683}.
CC -!- FUNCTION: PI-TfuII is a highly active homing endonuclease using Mg(2+)
CC as cofactor. Its minimal recognition and cleavage site is 21 bp (5'-
CC ACGCGGATACAGACGGCTTTT-3') cutting after -7 on both linear or circular
CC DNA substrates. Yields 5'-phosphate and 3'-OH ends. Its endonuclease
CC activity is strongly inhibited by the 3' digestion product, which
CC remains bound to the enzyme after the cleavage reaction.
CC {ECO:0000269|PubMed:10644683}.
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=2.7.7.7;
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:10644683};
CC Note=For PI-TfuII intein endonuclease. {ECO:0000269|PubMed:10644683};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:10644683};
CC Note=For PI-TfuI intein endonuclease. {ECO:0000269|PubMed:10644683};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Temperature dependence:
CC Optimum temperature is 70 degrees Celsius.;
CC -!- PTM: This protein undergoes a protein self splicing that involves a
CC post-translational excision of the two intervening regions (inteins)
CC followed by peptide ligation. {ECO:0000269|PubMed:10644683}.
CC -!- SIMILARITY: Belongs to the DNA polymerase type-B family. {ECO:0000305}.
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DR EMBL; Z69882; CAA93738.1; -; Genomic_DNA.
DR AlphaFoldDB; P74918; -.
DR REBASE; 4500; PI-TfuI.
DR REBASE; 4501; PI-TfuII.
DR PRIDE; P74918; -.
DR BRENDA; 2.7.7.7; 6299.
DR GO; GO:0003677; F:DNA binding; IEA:UniProtKB-KW.
DR GO; GO:0003887; F:DNA-directed DNA polymerase activity; IEA:UniProtKB-KW.
DR GO; GO:0004519; F:endonuclease activity; IEA:UniProtKB-KW.
DR GO; GO:0000166; F:nucleotide binding; IEA:InterPro.
DR GO; GO:0006260; P:DNA replication; IEA:UniProtKB-KW.
DR GO; GO:0016539; P:intein-mediated protein splicing; IEA:InterPro.
DR GO; GO:0006314; P:intron homing; IEA:UniProtKB-KW.
DR Gene3D; 1.10.132.60; -; 1.
DR Gene3D; 3.10.28.10; -; 2.
DR Gene3D; 3.30.420.10; -; 1.
DR Gene3D; 3.90.1600.10; -; 2.
DR InterPro; IPR006172; DNA-dir_DNA_pol_B.
DR InterPro; IPR006133; DNA-dir_DNA_pol_B_exonuc.
DR InterPro; IPR006134; DNA-dir_DNA_pol_B_multi_dom.
DR InterPro; IPR043502; DNA/RNA_pol_sf.
DR InterPro; IPR042087; DNA_pol_B_thumb.
DR InterPro; IPR023211; DNA_pol_palm_dom_sf.
DR InterPro; IPR003586; Hint_dom_C.
DR InterPro; IPR003587; Hint_dom_N.
DR InterPro; IPR036844; Hint_dom_sf.
DR InterPro; IPR027434; Homing_endonucl.
DR InterPro; IPR006142; INTEIN.
DR InterPro; IPR030934; Intein_C.
DR InterPro; IPR004042; Intein_endonuc.
DR InterPro; IPR006141; Intein_N.
DR InterPro; IPR004860; LAGLIDADG_2.
DR InterPro; IPR012337; RNaseH-like_sf.
DR InterPro; IPR036397; RNaseH_sf.
DR Pfam; PF00136; DNA_pol_B; 2.
DR Pfam; PF03104; DNA_pol_B_exo1; 2.
DR Pfam; PF14528; LAGLIDADG_3; 2.
DR PRINTS; PR00379; INTEIN.
DR SMART; SM00305; HintC; 2.
DR SMART; SM00306; HintN; 2.
DR SMART; SM00486; POLBc; 1.
DR SUPFAM; SSF51294; SSF51294; 2.
DR SUPFAM; SSF53098; SSF53098; 1.
DR SUPFAM; SSF55608; SSF55608; 2.
DR SUPFAM; SSF56672; SSF56672; 2.
DR TIGRFAMs; TIGR01443; intein_Cterm; 2.
DR TIGRFAMs; TIGR01445; intein_Nterm; 1.
DR PROSITE; PS50818; INTEIN_C_TER; 2.
DR PROSITE; PS50819; INTEIN_ENDONUCLEASE; 2.
DR PROSITE; PS50817; INTEIN_N_TER; 2.
PE 1: Evidence at protein level;
KW Autocatalytic cleavage; DNA replication; DNA-binding;
KW DNA-directed DNA polymerase; Endonuclease; Hydrolase; Intron homing;
KW Magnesium; Manganese; Nuclease; Nucleotidyltransferase; Protein splicing;
KW Repeat; Transferase.
FT CHAIN 1..406
FT /note="DNA polymerase, 1st part"
FT /id="PRO_0000007348"
FT CHAIN 407..766
FT /note="Homing endonuclease PI-TfuI"
FT /id="PRO_0000007349"
FT CHAIN 767..900
FT /note="DNA polymerase, 2nd part"
FT /id="PRO_0000007350"
FT CHAIN 901..1282
FT /note="Homing endonuclease PI-TfuII"
FT /id="PRO_0000007351"
FT CHAIN 1283..1523
FT /note="DNA polymerase, 3rd part"
FT /id="PRO_0000007352"
FT DOMAIN 524..665
FT /note="DOD-type homing endonuclease 1"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00273"
FT DOMAIN 1047..1186
FT /note="DOD-type homing endonuclease 2"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00273"
SQ SEQUENCE 1523 AA; 175918 MW; 7A2AC8236BF2E5F5 CRC64;
MILDTDYITE DGRPVIRVFK KENGEFKIEY DRDFEPYIYA LLKDDSAIED VKKITASRHG
TTVRVVRAGK VKKKFLGRPI EVWKLYFTHP QDVPAIRDKI REHPAVVDIY EYDIPFAKRY
LIDKGLIPME GDEELKMLAF DIETLYHEGE EFAEGPILMI SYADEEGARV ITWKKIDLPY
VDVVSTEKEM IKRFLKVVKE KDPDVLITYN GDNFDFAYLK KRSEKLGVKF ILGRDGSEPK
IQRMGDRFAV EVKGRIHFDL YPVIRHTINL PTYTLEAVYE AIFGQPKEKV YAEEIAQAWE
TGEGLERVAR YSMEDAKVTY ELGREFFPME AQLSRLVGQS FWDVSRSSTG NLVEWYLLRK
AYERNELAPN KPSGRELERR RGGYAGGYVK EPERGLWENI AYLDFRCHPA DTKVIVKGKG
VVNISEVREG DYVLGIDGWQ KVQRVWEYDY EGELVNINGL KCTPNHKLPV VRRTERQTAI
RDSLAKSFLT KKVKGKLITT PLFEKIGKIE REDVPEEEIL KGELAGIILA EGTLLRKDVE
YFDSSRGKKR VSHQYRVEIT VGAQEEDFQR RIVYIFERLF GVTPSVYRKK NTNAITFKVA
KKEVYLRVRE IMDGIENLHA PSVLRGFFEG DGSVNKVRKT VVVNQGTNNE WKIEVVSKLL
NKLGIPHRRY TYDYTEREKT MTTHILEIAG RDGLILFQTI VGFISTEKNM ALEEAIRNRE
VNRLENNAFY TLADFTAKTE YYKGKVYDLT LEGTPYYFAN GILTHNSLYP SIIISHNVSP
DTLNREGCGE YDEAPQVGHR FCKDFPGFIP SLLGDLLDER QKVKKHMKAT VDPIEKKLLD
YRQRAIKILA NSFYGYYGYA KARWYCKECA ESVTAWGRQY IETTMREIEE KFGFKVLYAD
SVTGDTEVTI RRNGRIEFVP IEKLFERVDH RVGEKEYCVL GGVEALTLDN RGRLVWKKVP
YVMRHKTDKR IYRVWFTNSW YLDVTEDHSL IGYLNTSKVK PGKPLKERLV EVKPEELGGK
VKSLITPNRP IARTIKANPI AVKLWELIGL LVGDGNWGGQ SNWAKYYVGL SCGLDKAEIE
RKVLNPLREA SVISNYYDKS KKGDVSILSK WLAGFMVKYF KDENGNKAIP SFMFNLPREY
IEAFLRGLFS ADGTVSLRRG IPEIRLTSVN RELSDAVRKL LWLVGVSNSL FTETKPNRYL
EKESGTHSIH VRIKNKHRFA DRIGFLIDRK STKLSENLGG HTNKKRAYKY DFDLVYPRKI
EEITYDGYVY DIEVEGTHRF FANGILVHNT DGFFATIPGA DAETVKKKAR EFLNYINPKL
PGLLELEYEG FYRRGFFVTK KKYAVIDEEG KITTRGLEIV RRDWSEVAKE TQARVLEAIL
RHGDVEEAVR IVKEVTEKLS KYEVPPEKLV IHEQITRELK DYKATGPHVA IAKRLAARGI
KVRPGTVISY IVLKGSGRIG DRTIPFDEFD PTKHRYDAEY YIENQVLPAV ERILKAFGYK
KEDLRYQKTR QVGLGAWLKM GKK
