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FANCD2 directly inhibits DNA2 nuclease at stalled replication forks and acts as a RAD51 mediator in strand exchange

Liu, Wenpeng and Roubal, Ivan and Polaczek, Piotr and Meng, Yuan and Choe, Won-chae and Caron, Marie-Christine and Sedgeman, Carl A. and Xi, Yu and Liu, Changwei and Wu, Qiong and Zheng, Li and Masson, Jean-Yves and Shen, Binghui and Campbell, Judith L. (2021) FANCD2 directly inhibits DNA2 nuclease at stalled replication forks and acts as a RAD51 mediator in strand exchange. . (Unpublished) https://resolver.caltech.edu/CaltechAUTHORS:20210712-151534393

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Abstract

FANCD2 protein, a key coordinator and effector of the interstrand crosslink repair pathway, is also required to prevent excessive nascent strand degradation at hydroxyurea induced stalled forks. The mechanisms of the fork protection are not well studied. Here, we purified FANCD2 to study how FANCD2 regulates DNA resection at stalled forks. In vitro, we showed that FANCD2 inhibits fork degradation in two ways: 1) it inhibits DNA2 nuclease activity by directly binding to DNA2. 2) independent of dimerization with FANCI, FANCD2 itself stabilizes RAD51 filaments to inhibit various nucleases, including DNA2. More unexpectedly, FANCD2 acts as a RAD51 mediator to stimulate the strand exchange activity of RAD51, and does so by enhancing ssDNA binding of RAD51. Our work biochemically explains mechanisms by which FANCD2 protects stalled forks and further provides a simple molecular explanation for genetic interactions between FANCD2 and the BRCA2 mediator.


Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription
https://doi.org/10.1101/2021.07.08.450798DOIDiscussion Paper
ORCID:
AuthorORCID
Shen, Binghui0000-0002-4408-407X
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. This version posted July 9, 2021. This work was supported by a CIHR foundation grant (J.-Y.M.) and J.-Y.M. is a FRQS Chair in genome stability; Korean government grants NRF-2017R1A2B2002289 and RF-2018R1A6A1A0302514 for W.C. sabbatical funding; R50CA211397 to L.Z.; R011CA085344 to B.S.; and USPS grant GM123554 to JLC. Author Contributions: Conceptualization, J.C.; Methodology, W.L., I.R., P.P., Y.M., M.C., Y.X., C.L., Q.W., L.Z.; Writing – Original Draft, W.L., P.P., J.C.; Writing – Review and Editing, W.L., L.Z., J.M., B.S, J.C.; Funding Acquisition, W.C., J.M., B.S., J.C; Supervision J.M., B.S., J.C. The authors declare no competing interests.
Funders:
Funding AgencyGrant Number
Canadian Institutes of Health Research (CIHR)UNSPECIFIED
Fonds de recherche du QuébecUNSPECIFIED
National Research Foundation of KoreaNRF-2017R1A2B2002289
National Research Foundation of KoreaNRF-2018R1A6A1A0302514
NIHR50CA211397
NIHR011CA085344
NIHGM123554
Record Number:CaltechAUTHORS:20210712-151534393
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20210712-151534393
Official Citation:FANCD2 directly inhibits DNA2 nuclease at stalled replication forks and acts as a RAD51 mediator in strand exchange. Wenpeng Liu, Ivan Roubal, Piotr Polaczek, Yuan Meng, Won-chae Choe, Marie- Christine Caron, Carl A. Sedgeman, Yu Xi, Changwei Liu, Qiong Wu, Li Zheng, Jean-Yves Masson, Binghui Shen, Judith L. Campbell. bioRxiv 2021.07.08.450798; doi: https://doi.org/10.1101/2021.07.08.450798
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:109775
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:12 Jul 2021 17:36
Last Modified:12 Jul 2021 17:36

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