The PCNA from Thermococcus fumicolans functionally interacts with DNA polymerase delta

Type Article
Date 2000-09
Language English
Author(s) Henneke Ghislaine, Raffin Jean-Paul, Ferrari Elena, Jonsson Zophonías, Dietrich Jacques, Hubscher Ulrich
Affiliation(s) IFREMER, Lab Biotechnol Microorganismes Hydrothermaux, DRV, VP, F-29280 Plouzane, France.
Univ Zurich, Inst Vet Biochem, CH-8057 Zurich, Switzerland.
Source Biochemical and Biophysical Research Communications (0006-291X) (Elsevier), 2000-09 , Vol. 276 , N. 2 , P. 600-606
DOI 10.1006/bbrc.2000.3481
WOS© Times Cited 16
Keyword(s) DNA polymerase delta, Conservation, Thermococcus fumicolans, Archaea, Hyperthermophile, PCNA
Abstract 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.
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Henneke Ghislaine, Raffin Jean-Paul, Ferrari Elena, Jonsson Zophonías, Dietrich Jacques, Hubscher Ulrich (2000). The PCNA from Thermococcus fumicolans functionally interacts with DNA polymerase delta. Biochemical and Biophysical Research Communications, 276(2), 600-606. Publisher's official version : https://doi.org/10.1006/bbrc.2000.3481 , Open Access version : https://archimer.ifremer.fr/doc/00000/659/