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The thermophilic crenarchaeon  Sulfolobus acidocaldarius  has four DNA polymerases (DNAPs): PolB1, PolB2, PolB3, and Dbh (PolY). Previous  in vitro  studies suggested that PolB1 is the main replicative DNAP of  Sulfolobales  whereas PolB2 and Y-family polymerases Dpo4 ( Saccharolobus solfataricus ) or Dbh are involved in DNA repair and translesion DNA synthesis. On the other hand, there are various opinions about the role of PolB3, which remains to be clearly resolved. In order to examine the roles of the DNAPs of  S. acidocaldarius  through  in vivo  experiments, we constructed  polB2 ,  polB3 , and  dbh  deletion strains and characterized their phenotypes. Efforts to construct a  polB1  deletion strain were not successful; in contrast, it was possible to isolate triple gene-deletion strains lacking  polB2 ,  polB3 , and  dbh . The growth of these strains was nearly the same as that of the parent strains under normal growth conditions. The  polB2 ,  polB3 , and  dbh  single-deletion strains were sensitive to some types of DNA-damaging treatments, but exhibited normal sensitivity to UV irradiation and several other damaging treatments. Overall, the genotype which exhibited the greatest sensitivity to the DNA-damaging treatments we tested was the Δ polB2  Δ polB3  combination, providing the first evidence of overlapping function for these two DNAPs  in vivo . The results of our study strongly suggest that PolB1 is responsible for the DNA replication of both the leading and lagging strands and is sufficient to complete the repair of most DNA damage under normal growth conditions in  S. acidocaldarius .

PolB1 Is Sufficient for DNA Replication and Repair Under Normal Growth Conditions in the Extremely Thermophilic Crenarchaeon Sulfolobus acidocaldarius