{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T22:37:52Z","timestamp":1770331072864,"version":"3.49.0"},"posted":{"date-parts":[[2026,1,13]]},"group-title":"Neuroscience","reference-count":79,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T00:00:00Z","timestamp":1768262400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"NIH","award":["R01NS091719"],"award-info":[{"award-number":["R01NS091719"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,1,13]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Mutations in the\n                  <jats:italic>leucine-rich repeat kinase 2<\/jats:italic>\n                  (\n                  <jats:italic>LRRK2<\/jats:italic>\n                  ) gene cause late-onset, autosomal dominant Parkinson\u2019s disease (PD).\n                  <jats:italic>LRRK2<\/jats:italic>\n                  encodes a multi-domain protein containing a Roc GTPase domain and a serine\/threonine-directed protein kinase domain, with PD-linked mutations known to enhance LRRK2 kinase activity and neuronal toxicity. Our previous studies identified the Golgi protein, ADP-Ribosylation Factor GTPase-Activating Protein 1 (ArfGAP1), as a novel modifier of LRRK2-induced cellular toxicity, where it can serve as a GAP-like protein and a robust kinase substrate of LRRK2. Here, we further explore the phosphorylation of ArfGAP1 by LRRK2 and its functional consequences. LRRK2 mediates the robust phosphorylation of ArfGAP1 within its lipid-sensing ALPS2 motif at residues Ser284, Thr291 and Thr292. We mutated these three phosphorylation sites, either alone or combined, to create hydrophobic phospho-null or charged phospho-mimicking versions of ArfGAP1. We find that modulating ArfGAP1 phosphorylation impairs its normal capacity to induce Golgi fragmentation upon overexpression in neural cells. Blocking phosphorylation impairs ArfGAP1-induced neurite outgrowth inhibition in primary neurons and protects against the neurotoxic effects of PD-linked G2019S LRRK2. ArfGAP1 interactome analysis in neural cells identifies 114 putative interacting proteins with a proportion of these unexpectedly localized to mitochondria, including the outer membrane proteins Voltage-Dependent Anion Channel (VDAC) 1-3. An ArfGAP1 triple phospho-mimic displays an increased interaction with mitochondrial VDACs owing to the redistribution of ArfGAP1 from the\n                  <jats:italic>cis<\/jats:italic>\n                  -Golgi to the cytoplasm. Promoting ArfGAP1 phosphorylation also blocks the formation of Golgi-derived vesicles following mild ER stress. Our data provides evidence for a complex functional interaction between LRRK2 and ArfGAP1 that serves to regulate ArfGAP1 subcellular localization, protein interactions, activity and neuronal toxicity via LRRK2-mediated phosphorylation of its membrane-binding ALPS2 motif. Our findings support additional validation of ArfGAP1 as a putative therapeutic target for modulating\n                  <jats:italic>LRRK2<\/jats:italic>\n                  -linked PD.\n                <\/jats:p>","DOI":"10.64898\/2026.01.12.699049","type":"posted-content","created":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T21:35:14Z","timestamp":1768340114000},"source":"Crossref","is-referenced-by-count":0,"title":["LRRK2 regulates ArfGAP1 membrane localization, activity and neuronal toxicity via phosphorylation within its lipid-sensing ALPS2 motif"],"prefix":"10.64898","author":[{"given":"Md. Shariful","family":"Islam","sequence":"first","affiliation":[{"name":"Department of Neurodegenerative Science, Van Andel Institute, Grand Rapids, Michigan 49503, United States"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valentin","family":"C\u00f3ppola-Segovia","sequence":"additional","affiliation":[{"name":"Department of Neurodegenerative Science, Van Andel Institute, Grand Rapids, Michigan 49503, United States"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessandra","family":"Musso","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Neurodegenerative Research, Brain Mind Institute, Swiss Federal Institute of Technology (EPFL), 1015 Lausanne, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2359-5679","authenticated-orcid":false,"given":"Darren J.","family":"Moore","sequence":"additional","affiliation":[{"name":"Department of Neurodegenerative Science, Van Andel Institute, Grand Rapids, Michigan 49503, United States"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2026020501235524000_2026.01.12.699049v1.1","doi-asserted-by":"publisher","DOI":"10.1002\/mds.26354"},{"key":"2026020501235524000_2026.01.12.699049v1.2","doi-asserted-by":"publisher","DOI":"10.1002\/mds.27115"},{"key":"2026020501235524000_2026.01.12.699049v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(19)30287-X"},{"key":"2026020501235524000_2026.01.12.699049v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2004.10.023"},{"key":"2026020501235524000_2026.01.12.699049v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2004.11.005"},{"key":"2026020501235524000_2026.01.12.699049v1.6","doi-asserted-by":"publisher","DOI":"10.1038\/ng.3955"},{"key":"2026020501235524000_2026.01.12.699049v1.7","doi-asserted-by":"publisher","DOI":"10.1038\/ng.3043"},{"key":"2026020501235524000_2026.01.12.699049v1.8","doi-asserted-by":"publisher","DOI":"10.1002\/mds.23265"},{"key":"2026020501235524000_2026.01.12.699049v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbadis.2008.09.015"},{"key":"2026020501235524000_2026.01.12.699049v1.10","doi-asserted-by":"publisher","DOI":"10.1007\/s11910-015-0564-y"},{"key":"2026020501235524000_2026.01.12.699049v1.11","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(08)70117-0"},{"key":"2026020501235524000_2026.01.12.699049v1.12","doi-asserted-by":"publisher","DOI":"10.1042\/BST20160264"},{"key":"2026020501235524000_2026.01.12.699049v1.13","doi-asserted-by":"crossref","unstructured":"Pfeffer, S.R. and D.R. 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