ID DPO4_CALS4 Reviewed; 384 AA. AC P58965; Q8RD00; DT 02-AUG-2002, integrated into UniProtKB/Swiss-Prot. DT 02-AUG-2002, sequence version 1. DT 02-OCT-2024, entry version 119. DE RecName: Full=DNA polymerase IV {ECO:0000255|HAMAP-Rule:MF_01113}; DE Short=Pol IV {ECO:0000255|HAMAP-Rule:MF_01113}; DE EC=2.7.7.7 {ECO:0000255|HAMAP-Rule:MF_01113, ECO:0000269|PubMed:37683741}; DE AltName: Full=Translesion synthesis DNA polymerase TTEDbh {ECO:0000303|PubMed:37683741}; DE Short=TLS DNA polymerase TTEDbh {ECO:0000303|PubMed:37683741}; GN Name=dinB {ECO:0000255|HAMAP-Rule:MF_01113}; GN OrderedLocusNames=TTE0254 {ECO:0000303|PubMed:37683741}; OS Caldanaerobacter subterraneus subsp. tengcongensis (strain DSM 15242 / JCM OS 11007 / NBRC 100824 / MB4) (Thermoanaerobacter tengcongensis). OC Bacteria; Bacillota; Clostridia; Thermoanaerobacterales; OC Thermoanaerobacteraceae; Caldanaerobacter. OX NCBI_TaxID=273068; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=DSM 15242 / JCM 11007 / NBRC 100824 / MB4; RX PubMed=11997336; DOI=10.1101/gr.219302; RA Bao Q., Tian Y., Li W., Xu Z., Xuan Z., Hu S., Dong W., Yang J., Chen Y., RA Xue Y., Xu Y., Lai X., Huang L., Dong X., Ma Y., Ling L., Tan H., Chen R., RA Wang J., Yu J., Yang H.; RT "A complete sequence of the T. tengcongensis genome."; RL Genome Res. 12:689-700(2002). RN [2] RP X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS) IN COMPLEX WITH MAGNESIUM, FUNCTION, RP CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, RP DOMAIN, MUTAGENESIS OF ASP-9; MET-10; ALA-12; PHE-13; PHE-14; ARG-38; RP SER-42; THR-43; TYR-46; ARG-49; ALA-56; ASP-103; LYS-147; LYS-154; GLY-180; RP GLY-182; SER-185; ILE-240; GLY-241; THR-245; PHE-290; LYS-297; THR-298; RP ARG-327 AND LEU-328, AND 3D-STRUCTURE MODELING IN COMPLEX WITH DOUBLE RP STRANDED DNA. RX PubMed=37683741; DOI=10.1016/j.ijbiomac.2023.126770; RA Tian L.F., Gao H., Yang S., Liu Y.P., Li M., Xu W., Yan X.X.; RT "Structure and function of extreme TLS DNA polymerase TTEDbh from RT Thermoanaerobacter tengcongensis."; RL Int. J. Biol. Macromol. 253:126770-126770(2023). CC -!- FUNCTION: Poorly processive, error-prone DNA polymerase involved in CC translesion repair and untargeted mutagenesis. Copies undamaged DNA at CC stalled replication forks, which arise in vivo from mismatched or CC misaligned primer ends. These misaligned primers can be extended by CC PolIV. Exhibits no 3'-5' exonuclease (proofreading) activity (By CC similarity). Involved in translesional synthesis. Primer extension CC fidelity in vitro is temperature-dependent. Inserts a correct base CC opposite templating bases at 70 degrees Celsius, but at 37 degrees CC Celsius in addition to correct base pairing, base transitions, CC transversions and frameshifts can occur. Preferably forms erroneous CC base pairs C:T. Bypasses 8-oxo-dG oxidative damage by incorporating CC dATP or dCTP opposite of the damaged DNA template site at both CC temperatures in vitro (PubMed:37683741). {ECO:0000250|UniProtKB:Q47155, CC ECO:0000255|HAMAP-Rule:MF_01113, ECO:0000269|PubMed:37683741}. 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; Evidence={ECO:0000255|HAMAP- CC Rule:MF_01113, ECO:0000269|PubMed:37683741}; CC -!- COFACTOR: CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; CC Evidence={ECO:0000255|HAMAP-Rule:MF_01113, CC ECO:0000269|PubMed:37683741}; CC Note=Binds 1 magnesium ion per subunit. {ECO:0000255|HAMAP- CC Rule:MF_01113, ECO:0000269|PubMed:37683741}; CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Temperature dependence: CC Optimum temperature is around 70 degrees Celsius for the DNA CC polymerase activity. Active also at 25 degrees Celsius, but CC polymerase activity increases with increasing temperature. Activity CC decreases with temperatures higher than 70 degrees Celsius. CC {ECO:0000303|PubMed:37683741}; CC -!- SUBUNIT: Monomer. {ECO:0000255|HAMAP-Rule:MF_01113, CC ECO:0000269|PubMed:37683741}. CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_01113}. CC -!- DOMAIN: The catalytic core consists of residues from subdomains of the CC finger (11-75), palm (77-161), thumb (163-224) and little finger (240- CC 335). {ECO:0000305|PubMed:37683741}. CC -!- SIMILARITY: Belongs to the DNA polymerase type-Y family. CC {ECO:0000255|HAMAP-Rule:MF_01113, ECO:0000303|PubMed:37683741}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; AE008691; AAM23550.1; -; Genomic_DNA. DR RefSeq; WP_009609988.1; NZ_JANUCV010000001.1. DR PDB; 7YLL; X-ray; 2.60 A; A=1-384. DR PDBsum; 7YLL; -. DR AlphaFoldDB; P58965; -. DR SMR; P58965; -. DR STRING; 273068.TTE0254; -. DR KEGG; tte:TTE0254; -. DR eggNOG; COG0389; Bacteria. DR HOGENOM; CLU_012348_1_2_9; -. DR OrthoDB; 9808813at2; -. DR Proteomes; UP000000555; Chromosome. DR GO; GO:0005737; C:cytoplasm; ISS:UniProtKB. DR GO; GO:0003887; F:DNA-directed DNA polymerase activity; IDA:UniProtKB. DR GO; GO:0003690; F:double-stranded DNA binding; IDA:UniProtKB. DR GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB. DR GO; GO:0032356; F:oxidized DNA binding; IDA:UniProtKB. DR GO; GO:0000731; P:DNA synthesis involved in DNA repair; IDA:UniProtKB. DR GO; GO:0006261; P:DNA-templated DNA replication; IDA:UniProtKB. DR GO; GO:0042276; P:error-prone translesion synthesis; IDA:UniProtKB. DR CDD; cd03586; PolY_Pol_IV_kappa; 1. DR Gene3D; 3.30.70.270; -; 1. DR Gene3D; 3.40.1170.60; -; 1. DR Gene3D; 1.10.150.20; 5' to 3' exonuclease, C-terminal subdomain; 1. DR Gene3D; 3.30.1490.100; DNA polymerase, Y-family, little finger domain; 1. DR HAMAP; MF_01113; DNApol_IV; 1. DR InterPro; IPR043502; DNA/RNA_pol_sf. DR InterPro; IPR036775; DNA_pol_Y-fam_lit_finger_sf. DR InterPro; IPR017961; DNA_pol_Y-fam_little_finger. DR InterPro; IPR050116; DNA_polymerase-Y. DR InterPro; IPR022880; DNApol_IV. DR InterPro; IPR053848; IMS_HHH_1. DR InterPro; IPR043128; Rev_trsase/Diguanyl_cyclase. DR InterPro; IPR001126; UmuC. DR PANTHER; PTHR11076:SF33; DNA POLYMERASE KAPPA; 1. DR PANTHER; PTHR11076; DNA REPAIR POLYMERASE UMUC / TRANSFERASE FAMILY MEMBER; 1. DR Pfam; PF00817; IMS; 1. DR Pfam; PF11799; IMS_C; 1. DR Pfam; PF21999; IMS_HHH_1; 1. DR SUPFAM; SSF56672; DNA/RNA polymerases; 1. DR SUPFAM; SSF100879; Lesion bypass DNA polymerase (Y-family), little finger domain; 1. DR PROSITE; PS50173; UMUC; 1. PE 1: Evidence at protein level; KW 3D-structure; Cytoplasm; DNA damage; DNA repair; DNA replication; KW DNA-binding; DNA-directed DNA polymerase; Magnesium; Metal-binding; KW Mutator protein; Nucleotidyltransferase; Reference proteome; Transferase. FT CHAIN 1..384 FT /note="DNA polymerase IV" FT /id="PRO_0000173960" FT DOMAIN 5..182 FT /note="UmuC" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01113" FT ACT_SITE 104 FT /evidence="ECO:0000255|HAMAP-Rule:MF_01113" FT BINDING 9 FT /ligand="Mg(2+)" FT /ligand_id="ChEBI:CHEBI:18420" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01113, FT ECO:0000269|PubMed:37683741, ECO:0007744|PDB:7YLL" FT BINDING 10 FT /ligand="Mg(2+)" FT /ligand_id="ChEBI:CHEBI:18420" FT /evidence="ECO:0000269|PubMed:37683741, FT ECO:0007744|PDB:7YLL" FT BINDING 103 FT /ligand="Mg(2+)" FT /ligand_id="ChEBI:CHEBI:18420" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01113, FT ECO:0000269|PubMed:37683741, ECO:0007744|PDB:7YLL" FT SITE 14 FT /note="Substrate discrimination" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01113" FT MUTAGEN 9 FT /note="D->A: Nearly complete loss of primer extension FT activity. No formation of correct or incorrect base pairs FT with the DNA template at 37 degrees Celsius. No FT incorporation of dATP, dGTP, dCTP, dTTP or dNTP mixes FT opposite 8-oxo-dG to bypass oxidative damage of the DNA FT template at neither 37 nor 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 10 FT /note="M->A: Increased primer extension activity compared FT to wild-type. Increased formation of incorrect base pairs FT with the DNA template at 37 degrees Celsius. Increased FT incorporation of dNTPs, especially dTTP, opposite 8-oxo-dG FT to bypass oxidative damage of the DNA template at 37 FT degrees Celsius, but shows no difference from wild-type at FT 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 12 FT /note="A->E: Decreased primer extension activity compared FT to wild-type. Forms correct, but not incorrect base pairs FT with the DNA template at 37 degrees Celsius. Can only FT incorporate dATP opposite 8-oxo-dG to bypass oxidative FT damage of the DNA template at 37 degrees Celsius, but FT complete loss of the bypass ability at 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 13 FT /note="F->A: Increased primer extension activity compared FT to wild-type. Increased formation of incorrect base pairs FT with the DNA template at 37 degrees Celsius. No difference FT from wild-type on the incorporation of dNTPs opposite 8- FT oxo-dG to bypass oxidative damage of the DNA template at 37 FT degrees Celsius, but partial loss of the bypass ability at FT 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 14 FT /note="F->A: Decreased primer extension activity compared FT to wild-type. Forms correct, but not incorrect base pairs FT with the DNA template at 37 degrees Celsius. Can only FT incorporate dATP opposite 8-oxo-dG to bypass oxidative FT damage of the DNA template at 37 degrees Celsius, but FT complete loss of the bypass ability at 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 38 FT /note="R->A: 20-fold lower dsDNA-binding compared to wild- FT type. Bypasses 8-oxo-dG oxidative damage of the DNA FT template at 37 degrees Celsius, but complete loss of the FT bypass ability at 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 42 FT /note="S->A: Decreased primer extension activity compared FT to wild-type. Loss of dCTP incorporation opposite adenine, FT but retained ability to form other incorrect base pairs FT with the DNA template at 37 degrees Celsius. No difference FT from wild-type on the incorporation of dNTPs opposite 8- FT oxo-dG to bypass oxidative damage of the DNA template at 37 FT degrees Celsius, but partial loss of the bypass ability at FT 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 43 FT /note="T->A: Decreased primer extension activity compared FT to wild-type. Partially retained formation of incorrect FT base pairs with the DNA template at 37 degrees Celsius. No FT difference from wild-type on the incorporation of dNTPs FT opposite 8-oxo-dG to bypass oxidative damage of the DNA FT template at 37 degrees Celsius, but partial loss of the FT bypass ability at 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 46 FT /note="Y->A: Decreased primer extension activity compared FT to wild-type. Forms correct, but not incorrect base pairs FT with the DNA template at 37 degrees Celsius. No difference FT from wild-type on the incorporation of dNTPs opposite 8- FT oxo-dG to bypass oxidative damage of the DNA template at 37 FT degrees Celsius, but complete loss of the bypass ability at FT 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 49 FT /note="R->A: Decreased primer extension activity compared FT to wild-type. Forms correct, but not incorrect base pairs FT with the DNA template at 37 degrees Celsius. Can only FT incorporate dATP opposite 8-oxo-dG to bypass oxidative FT damage of the DNA template at 37 degrees Celsius, but FT complete loss of the bypass ability at 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 56 FT /note="A->E: 20-fold lower dsDNA-binding compared to wild- FT type. Bypasses 8-oxo-dG oxidative damage of the DNA FT template at 37 degrees Celsius, but partial loss of the FT bypass ability at 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 103 FT /note="D->A: Decreased primer extension activity compared FT to wild-type. Forms correct base pairs to an extent with FT adenine, cytosine and thymine of the DNA template at 37 FT degrees Celsius. No incorporation of dATP, dGTP, dCTP, dTTP FT or dNTP mixes opposite 8-oxo-dG to bypass oxidative damage FT of the DNA template at neither 37 nor 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 147 FT /note="K->A: 20-fold lower dsDNA-binding compared to wild- FT type." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 154 FT /note="K->A: Significantly decreased primer extension FT activity compared to wild-type. Forms correct, but not FT incorrect base pairs with the DNA template at 37 degrees FT Celsius. No incorporation of dATP, dGTP, dCTP, dTTP or dNTP FT mixes opposite 8-oxo-dG to bypass oxidative damage of the FT DNA template at neither 37 nor 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 180 FT /note="G->V: 20-fold lower dsDNA-binding compared to wild- FT type." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 182 FT /note="G->E: Significantly decreased dsDNA-binding compared FT to wild-type." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 185 FT /note="S->A: Significantly decreased dsDNA-binding compared FT to wild-type." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 240 FT /note="I->A: 20-fold lower dsDNA-binding compared to wild- FT type; when associated with E-241." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 241 FT /note="G->E: 20-fold lower dsDNA-binding compared to wild- FT type; when associated with A-240." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 245 FT /note="T->A: Significantly decreased dsDNA-binding compared FT to wild-type." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 290 FT /note="F->A: Significantly decreased dsDNA-binding compared FT to wild-type. Bypasses 8-oxo-dG oxidative damage of the DNA FT template at 37 degrees Celsius, but partial loss of the FT bypass ability at 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 297 FT /note="K->A: 20-fold lower dsDNA-binding compared to wild- FT type; when associated with A-298." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 298 FT /note="T->A: 20-fold lower dsDNA-binding compared to wild- FT type; when associated with A-297." FT /evidence="ECO:0000269|PubMed:37683741" FT MUTAGEN 327 FT /note="R->A: Significantly decreased dsDNA-binding compared FT to wild-type. Bypasses 8-oxo-dG oxidative damage of the DNA FT template at 37 degrees Celsius, but complete loss of the FT bypass ability at 70 degrees Celsius." FT /evidence="ECO:0000269|PubMed:37683741" SQ SEQUENCE 384 AA; 43916 MW; 01C214516AD9AABB CRC64; MKRKIIHVDM DAFFASIEQQ DNPEYRGKPV IVGGLSGRGV VSTCSYEARK YGIHSAMPMY MAKKLCPQGI FLPVRRKRYE EVSEQIFRIL YDITPFVEPV SIDEAYLDVT HVDKNPEDIA LEIKKRVKDA TGLTVSVGIS YNKFLAKLAS DWNKPDGLMV ITEDMVPEIL KPLPVTKVHG IGEKSAEKLR SIGIETVEDL LKLPQENLIE LFGKTGVEIY NRIRGIDERP VETMREIKSI GKEKTLEKDT KNKELLIQHL KEFSEIVSEE LIKERLYCRT VTVKIKTADF AVHTKSKTVD KYIRFSEDIY EVAKGILEEW KLEQYVRLIG LSVSNLSPVK YEQLSFLDKR LVKVIKAGNL AEEINKRIGK KIIKKGSELL KDNK //