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| Mesiet Cladiere, L; Norais, Cédric; Kuhn, Joelle; Briffotaux, Julien; Sloostra, Jerry W.; Ferrari, Elena; Hubscher, Ulrich; Flament, Didier; Myllykallio, Hannu. |
| During DNA replication and repair, many proteins bind to and dissociate in a highly specific and ordered manner from proliferating cell nuclear antigen (PCNA). We describe a combined approach of in silico searches at the genome level and combinatorial peptide synthesis to investigate the binding properties of hundreds of short PCNA-interacting peptides (PIP-peptides) to archaeal and eukaryal PCNAs. Biological relevance of our combined approach was demonstrated by identification an inactive complex of Pyrococcus abyssi ribonuclease HII with PCNA. Furthermore we show that PIP-peptides interact with PCNA largely in a sequence independent manner. Our experimental approach also identified many so far unidentified PCNA interacting peptides in a number of human... |
| Tipo: Text |
Palavras-chave: Ribonuclease HII; Combinatorial peptide synthesis; PCNA. |
| Ano: 2007 |
URL: http://archimer.ifremer.fr/doc/2007/publication-3297.pdf |
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| Henneke, Ghislaine; Raffin, Jean-paul; Ferrari, Elena; Jonsson, Zophonías; Dietrich, Jacques; Hubscher, Ulrich. |
| 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.... |
| Tipo: Text |
Palavras-chave: DNA polymerase delta; Conservation; Thermococcus fumicolans; Archaea; Hyperthermophile; PCNA. |
| Ano: 2000 |
URL: http://archimer.ifremer.fr/doc/2000/publication-659.pdf |
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