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Physical interaction between proliferating cell nuclear antigen and replication factor C from Pyrococcus furiosus
Author(s) -
Matsumiya Shigeki,
Ishino Sonoko,
Ishino Yoshizumi,
Morikawa Kosuke
Publication year - 2002
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1046/j.1365-2443.2002.00572.x
Subject(s) - pyrococcus furiosus , proliferating cell nuclear antigen , biology , dna replication , dna polymerase , dna clamp , dna , replication factor c , protein subunit , microbiology and biotechnology , biophysics , biochemistry , reverse transcriptase , gene , eukaryotic dna replication , archaea , rna
Background: Proliferating cell nuclear antigen (PCNA), which is recognized as a DNA polymerase processivity factor, has direct interactions with various proteins involved in the important genetic information processes in Eukarya. We determined the crystal structure of PCNA from the hyperthermophilic archaeon, Pyrococcus furiosus ( Pfu PCNA) at 2.1 Å resolution, and found that the toroidal ring‐shaped structure, which consists of homotrimeric molecules, is highly conserved between the Eukarya and Archaea. This allowed us to examine its interaction with the loading factor at the atomic level. Results: The replication factor C (RFC) is known as the loading factor of PCNA on to the DNA strand. P. furiosus RFC ( Pfu RFC) has a PCNA binding motif (PIP‐box) at the C‐terminus of the large subunit (RFCL). An 11 residue‐peptide containing a PIP‐box sequence of RFCL inhibited the PCNA‐dependent primer extension ability of P. furiosus Pol I in a concentration‐dependent manner. To understand the molecular interaction mechanism of PCNA with PCNA binding proteins, we solved the crystal structure of Pfu PCNA complexed with the PIP‐box peptide. The interaction mode of the two molecules is remarkably similar to that of human PCNA and a peptide containing the PIP‐box of p21 WAF1/CIP1 . Moreover, the PIP‐box binding may have some effect on the stability of the ring structure of Pfu PCNA by some domain shift. Conclusions: Our structural analysis on Pfu PCNA suggests that the interaction mode of the PIP‐box with PCNA is generally conserved among the PCNA interacting proteins and that the functional meaning of the interaction via the PIP‐box possibly depends on each protein. A movement of the C‐terminal region of the PCNA monomer by PIP‐box binding may cause the PCNA ring to be more rigid, suitable for its functions.