| 1. |
Ryo Fujisawa, Eiji Ohashi, Kouji Hirota, Toshiki Tsurimoto, Human CTF18-RFC clamp-loader complexed with non-synthesising DNA polymerase ϵ efficiently loads the PCNA sliding clamp, Nucleic acids research, 10.1093/nar/gkx096, 45, 8, 4550-4563, 2017.01, The alternative proliferating-cell nuclear antigen (PCNA)-loader CTF18-RFC forms a stable complex with DNA polymerase ϵ (Polϵ). We observed that, under near-physiological conditions, CTF18- RFC alone loaded PCNA inefficiently, but loaded it efficiently when complexed with Polϵ. During efficient PCNA loading, CTF18-RFC and Polϵ assembled at a 3ϵ primer-template junction cooperatively, and directed PCNA to the loading site. Site-specific photo-crosslinking of directly interacting proteins at the primer-template junction showed similar cooperative binding, in which the catalytic N-terminal portion of Polϵ acted as the major docking protein. In the PCNA-loading intermediate with ATPαS, binding of CTF18 to the DNA structures increased, suggesting transient access of CTF18-RFC to the primer terminus. Polϵ placed in DNA synthesis mode using a substrate DNA with a deoxidised 3ϵ primer end did not stimulate PCNA loading, suggesting that DNA synthesis and PCNA loading are mutually exclusive at the 3ϵ primer-template junction. Furthermore, PCNA and CTF18-RFC-Polϵ complex engaged in stable trimeric assembly on the template DNA and synthesised DNA efficiently. Thus, CTF18-RFC appears to be involved in leading-strand DNA synthesis through its interaction with Polϵ, and can load PCNA onto DNA when Polϵ is not in DNA synthesis mode to restore DNA synthesis.. |
| 2. |
Okimoto H, Tanaka S, Araki H, Ohashi E, Tsurimoto T., Conserved interaction of Ctf18-RFC with DNA polymerase ε is critical for maintenance of genome stability in Saccharomyces cerevisiae., Genes to Cells, 21: 482-491, 2016.05. |
| 3. |
Takeishi Y, Iwaya-Omi R, Ohashi E, Tsurimoto T, Intramolecular binding of the Rad9 C terminus in the checkpoint clamp Rad9-Hus1-Rad1 is closely linked with its DNA binding., Journal of Biological Chemistry, 19923-19932, 290: 19923-19932, 2015.08. |
| 4. |
Ohashi E, Takeishi Y, Ueda S, Tsurimoto T, Interaction between Rad9-Hus1-Rad1 and TopBP1 activates ATR-ATRIP and promotes TopBP1 recruitment to sites of UV-damage., DNA repair, 1-11, 21: 1-11, 2014.09. |
| 5. |
Ueda S, Takeishi Y, Ohashi E, Tsurimoto T., Two serine phosphorylation sites in the C-terminus of Rad9 are critical for 9-1-1 binding to TopBP1 and activation of the DNA damage checkpoint response in HeLa cells., Genes to Cells, 807-816, 17: 807-816, 2012.10. |
| 6. |
Takeishi Y, Ohashi E, Ogawa K, Masai H, Obuse C, Tsurimoto T., Casein kinase 2-dependent phosphorylation of human Rad9 mediates the interaction between human Rad9-Hus1-Rad1 complex and TopBP1., Genes to Cells, 15: 761-771, 2010.06. |
| 7. |
Ohashi E, Hanafusa T, Kamei K, Song I, Tomida J, Hashimoto H, Vaziri C, Ohmori H., Identification of a novel REV1-interacting motif necessary for DNA polymerase kappa function., Genes to Cells, 14:101-11, 2009.02. |
| 8. |
Ohashi E, Murakumo Y, Kanjo N, Akagi J, Masutani C, Hanaoka F, Ohmori H., Interaction of hREV1 with three human Y-family DNA polymerases., Genes to Cells, 523-531, 9: 523-531, 2004.06. |
| 9. |
Ohashi E, Bebenek K, Matsuda T, Feaver WJ, Gerlach VL, Friedberg EC, Ohmori H, Kunkel TA., Fidelity and processivity of DNA synthesis by DNA polymerase kappa, the product of the human DINB1 gene., Journal of Biological Chemistry, 39678-39684, 275: 39678-39684, 2000.12. |
| 10. |
Ohashi E, Ogi T, Kusumoto R, Iwai S, Masutani C, Hanaoka F, Ohmori H., Error-prone bypass of certain DNA lesions by the human DNA polymerase kappa., Genes and Development, 1589-1594, 14: 1589-1594, 2000.07. |
