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Characterization of a Bacillus subtilis 64-kDa DNA polymerase X potentially involved in DNA repair
Authors:Baños Benito  Lázaro José M  Villar Laurentino  Salas Margarita  de Vega Miguel
Institution:Instituto de Biología Molecular “Eladio Viñuela” (CSIC), Centro de Biología Molecular “Severo Ochoa” (CSIC-UAM), C/Nicolás Cabrera 1, Cantoblanco, 28049 Madrid, Spain
Abstract:Bacillus subtilis gene yshC encodes a 64-kDa family X DNA polymerase (PolXBs), which contains all the critical residues involved in DNA and nucleotide binding as well as those responsible for catalysis of DNA polymerization, conserved in most family X members. Biochemical analyses of the purified enzyme indicate that PolXBs is a monomeric and strictly template-directed DNA polymerase, preferentially acting on DNA structures containing gaps from one to a few nucleotides and bearing a phosphate group at the 5' end of the downstream DNA. The fact that PolXBs is able to conduct filling of a single-nucleotide gap, allowing further sealing of the resulting nick by a DNA ligase, points to a putative role in base excision repair during the B. subtilis life cycle.
Keywords:BSA  bovine serum albumin  TdT  terminal deoxynucleotidyl transferase  PolX  family X of DNA polymerases  BER  base excision repair  HhH  helix-hairpin-helix  dRP  5&prime  -deoxyribose 5&prime  -phosphate  ORF  open reading frame  EDTA  ethylenediaminetetraacetic acid  dNMP  deoxyribonucleoside 5&prime  -monophosphate  ddNTP  2&prime  -3&prime  -dideoxynucleoside 5&prime  -triphosphate
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