{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,4,12]],"date-time":"2026-04-12T05:49:00Z","timestamp":1775972940057,"version":"3.50.1"},"posted":{"date-parts":[[2025,5,18]]},"group-title":"Cancer Biology","reference-count":103,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,5,18]],"date-time":"2025-05-18T00:00:00Z","timestamp":1747526400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Cancer Institute","award":["R37CA227656"],"award-info":[{"award-number":["R37CA227656"]}]},{"name":"National Cancer Institute","award":["P30CA177558"],"award-info":[{"award-number":["P30CA177558"]}]},{"name":"Kentucky Pediatric Research Trust Fund"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,5,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Phosphatase of Regenerating Liver 3 (PRL-3) is frequently upregulated in cancer and associated with poor prognosis, yet its oncogenic mechanism remains unresolved. Although traditionally studied for its phosphatase activity, PRL-3 also engages in protein-protein interactions via its catalytic site, notably binding the CNNM magnesium transporters, rendering these functions mutually exclusive. The commonly used C104S mutant disrupts both activities, confounding interpretations of prior studies. To dissect PRL-3\u2019s distinct functions, we employed mutants selectively deficient in phosphatase activity (C104D) or CNNM binding (R138E). In zebrafish models of acute lymphoblastic leukemia (ALL) and rhabdomyosarcoma (RMS), as well as in human cancer cell lines, both wild-type PRL-3 and C104D enhanced tumor initiation, growth, and dissemination, while R138E had no effect. These findings indicate that PRL-3 promotes cancer via non-catalytic mechanisms. To support therapeutic development, we established an in vitro FRET-based assay to screen for inhibitors of the PRL-3:CNNM interaction and validated a cyclic peptide as a positive control. By demonstrating that PRL-3 enhances cancer progression independently of its catalytic activity, this study shifts focus toward targeting its binding functions as a therapeutic strategy.<\/jats:p>\n                <jats:sec>\n                  <jats:title>Graphical Abstract<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" orientation=\"portrait\" fig-type=\"figure\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"654016v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Statement of significance<\/jats:title>\n                  <jats:p>This study demonstrates that PRL-3 drives cancer initiation and progression through a phosphatase-independent mechanism across diverse models, highlighting the need to therapeutically target its non-catalytic protein interactions in future cancer treatment strategies.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.05.14.654016","type":"posted-content","created":{"date-parts":[[2025,5,18]],"date-time":"2025-05-18T23:40:11Z","timestamp":1747611611000},"source":"Crossref","is-referenced-by-count":0,"title":["Phosphatase activity is dispensable for PRL-3-mediated oncogenesis and tumor progression"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6748-3591","authenticated-orcid":false,"given":"Jeffery T.","family":"Jolly","sequence":"first","affiliation":[{"name":"Department of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky"},{"name":"Markey Cancer Center, University of Kentucky"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Majd A.","family":"Al-Hamaly","sequence":"additional","affiliation":[{"name":"Markey Cancer Center, University of Kentucky"},{"name":"Department of Pharmacology and Nutritional Sciences, College of Medicine, University of Kentucky"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Caroline N.","family":"Smith","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky"},{"name":"Markey Cancer Center, University of Kentucky"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H. 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