The PCNA from Thermococcus fumicolans functionally interacts with DNA polymerase delta

Biochem Biophys Res Commun. 2000 Sep 24;276(2):600-6. doi: 10.1006/bbrc.2000.3481.


We have cloned the gene encoding proliferating cell nuclear antigen (PCNA) from the hyperthermophilic euryarchaeote Thermococcus fumicolans (Tfu). Tfu PCNA contains 250 amino acids with a calculated M(r) of 28,000 and is 26% identical to human PCNA. Next, Tfu PCNA was overexpressed in Escherichia coli and it showed an apparent molecular mass of 33.5 kDa. The purified Tfu PCNA was tested first with recombinant Tfu DNA polymerase I (Tfu pol) and second with calf thymus DNA polymerase delta (pol delta). When tested with the homologous Tfu pol on bacteriophage lambda DNA, large amounts of Tfu PCNA were required to obtain two- to threefold stimulation. Surprisingly, however, Tfu PCNA was much more efficient than human PCNA in stimulating calf thymus pol delta. Our data suggest that PCNA has been functionally conserved not only within eukaryotes but also from hyperthermophilic euryarchaeotes to mammals.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Archaeal Proteins / biosynthesis
  • Archaeal Proteins / chemistry
  • Archaeal Proteins / genetics
  • Archaeal Proteins / pharmacology
  • Cattle
  • Cloning, Molecular
  • DNA Polymerase III / drug effects
  • DNA Polymerase III / metabolism*
  • Enzyme Activation
  • Escherichia coli
  • Humans
  • Molecular Sequence Data
  • Proliferating Cell Nuclear Antigen / biosynthesis
  • Proliferating Cell Nuclear Antigen / chemistry
  • Proliferating Cell Nuclear Antigen / genetics
  • Proliferating Cell Nuclear Antigen / pharmacology*
  • Protein Conformation
  • Recombinant Proteins / drug effects
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Thermococcus / chemistry*
  • Transfection


  • Archaeal Proteins
  • Proliferating Cell Nuclear Antigen
  • Recombinant Proteins
  • DNA Polymerase III