The hyperthermophilic euryarchaeota Pyrococcus abyssi likely requires the two DNA polymerases D and B for DNA replication.
DNA polymerases carry out DNA synthesis during DNA replication, DNA recombination and DNA repair. During the past five years, the number of DNA polymerases in both eukarya and bacteria has increased to at least 19 and multiple biological roles have been assigned to many DNA polymerases. Archaea, the third domain of life, on the other hand, have only a subset of the eukaryotic-like DNA polymerases. The diversity among the archaeal DNA polymerases poses the intriguing question of their functional tasks. Here, we focus on the two identified DNA polymerases, the family B DNA polymerase B (PabpolB) and the family D DNA polymerase D (PabpolD) from the hyperthermophilic euryarchaeota Pyrococcus abyssi. Our data can be summarized as follows: (i) both Pabpols are DNA polymerizing enzymes exclusively; (ii) their DNA binding properties as tested in gel shift competition assays indicated that PabpolD has a preference for a primed template; (iii) PabPolD is a primer-directed DNA polymerase independently of the primer composition whereas PabpolB behaves as an exclusively DNA primer-directed DNA polymerase; (iv) PabPCNA is required for PabpolD to perform efficient DNA synthesis but not PabpolB; (v) PabpolD, but not PabpolB, contains strand displacement activity; (vii) in the presence of PabPCNA, however, both Pabpols D and B show strand displacement activity; and (viii) we show that the direct interaction between PabpolD and PabPCNA is DNA-dependent. Our data imply that PabPolD might play an important role in DNA replication likely together with PabpolB, suggesting that archaea require two DNA polymerases at the replication fork.
Henneke
Ghislaine
G
IFREMER, UMR 6197, Laboratoire de Microbiologie et Environnements Extrêmes, DRV/VP/LM2E, BP 70, F-29280 Plouzané, France. ghenneke@ifremer.fr
Flament
Didier
D
Hübscher
Ulrich
U
Querellou
Joël
J
Raffin
Jean-Paul
JP
D017931:DNA Primers / Q000235:genetics; D004261:DNA Replication; D019641:DNA, Archaeal / Q000096:biosynthesis* / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D043903:Pyrococcus abyssi / Q000201:enzymology* / Q000235:genetics; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D013696:Temperature; D013698:Templates, Genetic
D017931:DNA Primers; D019641:DNA, Archaeal; D018809:Proliferating Cell Nuclear Antigen; D012313:RNA; D004259:DNA-Directed DNA Polymerase