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Kawasoe, Yoshitaka, Shimokawa, Sakiko, Gillespie, Peter J., Blow, J. Julian 
ORCID: https://orcid.org/0000-0002-9524-5849, Tsurimoto, Toshiki and Takahashi, Tatsuro S.
(2024)
The Atad5 RFC-like complex is the major unloader of proliferating cell nuclear antigen in Xenopus egg extracts.
Journal of Biological Chemistry, 300 (1).
ISSN 0021-9258
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Abstract
Proliferating cell nuclear antigen (PCNA) is a homo-trimeric clamp complex that serves as the molecular hub for various DNA transactions, including DNA synthesis and post-replicative mismatch repair. Its timely loading and unloading are critical for genome stability. PCNA loading is catalyzed by Replication factor C (RFC) and the Ctf18 RFC-like complex (Ctf18-RLC), and its unloading is catalyzed by Atad5/Elg1-RLC. However, RFC, Ctf18-RLC, and even some subcomplexes of their shared subunits are capable of unloading PCNA in vitro, leaving an ambiguity in the division of labor in eukaryotic clamp dynamics. By using a system that specifically detects PCNA unloading, we show here that Atad5-RLC, which accounts for only approximately 3% of RFC/RLCs, nevertheless provides the major PCNA unloading activity in Xenopus egg extracts. RFC and Ctf18-RLC each account for approximately 40% of RFC/RLCs, while immunodepletion of neither Rfc1 nor Ctf18 detectably affects the rate of PCNA unloading in our system. PCNA unloading is dependent on the ATP-binding motif of Atad5, independent of nicks on DNA and chromatin assembly, and inhibited effectively by PCNA-interacting peptides. These results support a model in which Atad5-RLC preferentially unloads DNA-bound PCNA molecules that are free from their interactors.
| Item Type: | Article |
|---|---|
| Additional Information: | Funding information: This work was supported by JSPS KAKENHI grants [17H06935, 19K16042 to Y. K., and 22H04697, 20H05392, 20H03186 to T. S. T.] and Wellcome Trust Senior Investigator Award WT096598MA to J. J. B. |
| Uncontrolled Keywords: | dna replication,xenopus egg extract,clamp loader,mismatch repair,proliferating cell nuclear antigen,replication clamp |
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| Depositing User: | LivePure Connector |
| Date Deposited: | 10 Jun 2024 11:30 |
| Last Modified: | 22 Aug 2024 11:34 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/95447 |
| DOI: | 10.1016/j.jbc.2023.105588 |
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