{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:40:52Z","timestamp":1785426052511,"version":"3.56.0"},"posted":{"date-parts":[[2026,5,24]]},"group-title":"Cell Biology","reference-count":57,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,24]],"date-time":"2026-05-24T00:00:00Z","timestamp":1779580800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R01GM144579"],"award-info":[{"award-number":["R01GM144579"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01GM136717"],"award-info":[{"award-number":["R01GM136717"]}],"id":[{"id":"https:\/\/ror.org\/04q48ey07","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01CA237286"],"award-info":[{"award-number":["R01CA237286"]}],"id":[{"id":"https:\/\/ror.org\/040gcmg81","id-type":"ROR","asserted-by":"publisher"}]},{"award":["P01CA275717"],"award-info":[{"award-number":["P01CA275717"]}],"id":[{"id":"https:\/\/ror.org\/040gcmg81","id-type":"ROR","asserted-by":"publisher"}]},{"award":["REI RR210023"],"award-info":[{"award-number":["REI RR210023"]}],"id":[{"id":"https:\/\/ror.org\/0003xa228","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000090","name":"Congressionally Directed Medical Research Programs","doi-asserted-by":"crossref","award":["BC191160"],"award-info":[{"award-number":["BC191160"]}],"id":[{"id":"10.13039\/100000090","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,24]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>Replication fork reversal alleviates DNA replication stress and maintains genome stability. We previously showed that FBH1 and RAD54L cooperate to promote fork reversal in human cells, and that FBH1-dependent fork reversal requires the branch migration activity of RAD54L. However, the molecular basis of this cooperation remained unclear.<\/jats:p>\n                <jats:p>Here, we identify both a physical and functional interaction between FBH1 and RAD54L. We demonstrate that purified FBH1 and RAD54L interact directly and form a complex at stalled replication forks in cells. Mapping studies revealed that RAD54L Lobe 1 is critical for interaction with the FBH1 2B subdomain. In cells, FBH1-RAD54L complex formation is enhanced in the absence of RAD51. Consistently, purified RAD54L displays a stronger affinity for RAD51 than for FBH1.<\/jats:p>\n                <jats:p>Using biochemical reconstitution assays, we further show that FBH1 and RAD54L promote fork reversal more efficiently together than either protein alone, with maximal reversal observed when FBH1 acts before RAD54L. Collectively, our findings establish RAD54L as an essential functional partner of FBH1 in replication fork reversal and provide mechanistic insight into the sequential coordination of their activities.<\/jats:p>","DOI":"10.64898\/2026.05.22.727240","type":"posted-content","created":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T01:55:14Z","timestamp":1779674114000},"source":"Crossref","is-referenced-by-count":0,"title":["FBH1 and RAD54L directly interact and cooperate to drive replication fork reversal"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4142-8260","authenticated-orcid":false,"given":"Mollie E.","family":"Uhrig","sequence":"first","affiliation":[{"name":"Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, CO, 80523, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Courtney N.","family":"Johnson","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Structural Biology, UT Health San Antonio, San Antonio, TX, 78229, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jordi","family":"Gomez","sequence":"additional","affiliation":[{"name":"Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, CO, 80523, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5430-9797","authenticated-orcid":false,"given":"Alexander V.","family":"Mazin","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Structural Biology, UT Health San Antonio, San Antonio, TX, 78229, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6844-4693","authenticated-orcid":false,"given":"Claudia","family":"Wiese","sequence":"additional","affiliation":[{"name":"Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, CO, 80523, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026052701100935000_2026.05.22.727240v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb2897"},{"key":"2026052701100935000_2026.05.22.727240v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/J.MOLCEL.2022.05.004"},{"key":"2026052701100935000_2026.05.22.727240v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2019.04.027"},{"key":"2026052701100935000_2026.05.22.727240v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.dnarep.2024.103731"},{"key":"2026052701100935000_2026.05.22.727240v1.5","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-062917-011921"},{"key":"2026052701100935000_2026.05.22.727240v1.6","doi-asserted-by":"publisher","DOI":"10.1038\/nrm3935"},{"key":"2026052701100935000_2026.05.22.727240v1.7","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201406099"},{"key":"2026052701100935000_2026.05.22.727240v1.8","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkab344"},{"key":"2026052701100935000_2026.05.22.727240v1.9","doi-asserted-by":"crossref","unstructured":"Liu, W. , Krishnamoorthy, A. , Zhao, R. and Cortez, D. 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