A novel proteomic approach identifies new interaction partners for proliferating cell nuclear antigen.
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 proteins.
Meslet-Cladiére
Laurence
L
CNRS UMR 8621 U. Paris-Sud, 91405 Orsay Cedex, France.
Norais
Cédric
C
Kuhn
Joëlle
J
Briffotaux
Julien
J
Sloostra
Jerry W
JW
Ferrari
Elena
E
Hübscher
Ulrich
U
Flament
Didier
D
Myllykallio
Hannu
H
D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D011088:DNA Ligases / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair; D045585:Flap Endonucleases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics / Q000378:metabolism*; D020543:Proteome; D043903:Pyrococcus abyssi / Q000201:enzymology; D016914:Ribonuclease H / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
D019843:Archaeal Proteins; D010455:Peptides; D018809:Proliferating Cell Nuclear Antigen; D020543:Proteome; D045585:Flap Endonucleases; C091139:ribonuclease HII; D016914:Ribonuclease H; D011088:DNA Ligases