{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:36:00Z","timestamp":1785404160831,"version":"3.56.0"},"posted":{"date-parts":[[2025,8,30]]},"group-title":"Biochemistry","reference-count":38,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,8,30]],"date-time":"2025-08-30T00:00:00Z","timestamp":1756512000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R01GM147332"],"award-info":[{"award-number":["R01GM147332"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01GM113164"],"award-info":[{"award-number":["R01GM113164"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["GM103310"],"award-info":[{"award-number":["GM103310"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["OD019994"],"award-info":[{"award-number":["OD019994"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000893","name":"Simons Foundation","doi-asserted-by":"crossref","award":["SF349247"],"award-info":[{"award-number":["SF349247"]}],"id":[{"id":"10.13039\/100000893","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100009344","name":"Agouron Institute","doi-asserted-by":"crossref","award":["F00316"],"award-info":[{"award-number":["F00316"]}],"id":[{"id":"10.13039\/100009344","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,8,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Pathogenic variants in the motor domain of the kinesin-3 motor protein KIF1A cause a range of neurodevelopmental and neurodegenerative conditions collectively termed KIF1A-associated neurological disorder (KAND). Among these, mutations at residue R350 are linked to hereditary spastic paraplegia and altered motor function. Yet, the structural basis for their pathogeny remains unclear. Here, we present high-resolution cryo-electron microscopy (cryo-EM) structures of KIF1A R350G and R350W bound to microtubules in both the apo and AMP-PNP-bound states. We identify a previously unrecognized salt bridge between KIF1A residue R350 and \u03b1-tubulin E415 that is disrupted in both mutants. This loss of electrostatic interaction correlates with increased velocity and reduced processivity, as demonstrated by single-molecule assays. Our results reveal a conserved electrostatic interaction at the motor\u2013microtubule interface that regulates KIF1A\u2019s motility behavior.<\/jats:p>","DOI":"10.1101\/2025.08.30.673262","type":"posted-content","created":{"date-parts":[[2025,8,31]],"date-time":"2025-08-31T05:45:13Z","timestamp":1756619113000},"source":"Crossref","is-referenced-by-count":1,"title":["Pathogenic KIF1A R350 Variants Disrupt A Conserved Kinesin-Tubulin Salt Bridge"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4953-9202","authenticated-orcid":false,"given":"Abhipsa","family":"Shatarupa","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7447-1484","authenticated-orcid":false,"given":"Lu","family":"Rao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3752-9453","authenticated-orcid":false,"given":"Ana B.","family":"Asenjo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8346-5473","authenticated-orcid":false,"given":"Arne","family":"Gennerich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0483-5369","authenticated-orcid":false,"given":"Hernando","family":"Sosa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025090317402788000_2025.08.30.673262v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(91)90391-B"},{"key":"2025090317402788000_2025.08.30.673262v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(95)90538-3"},{"key":"2025090317402788000_2025.08.30.673262v1.3","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.141.2.431"},{"key":"2025090317402788000_2025.08.30.673262v1.4","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0854.2004.00195.x"},{"key":"2025090317402788000_2025.08.30.673262v1.5","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E07-03-0261"},{"key":"2025090317402788000_2025.08.30.673262v1.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.neulet.2011.01.018"},{"key":"2025090317402788000_2025.08.30.673262v1.7","doi-asserted-by":"publisher","DOI":"10.1038\/nn.4213"},{"key":"2025090317402788000_2025.08.30.673262v1.8","doi-asserted-by":"publisher","DOI":"10.1038\/nn.2665"},{"key":"2025090317402788000_2025.08.30.673262v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.xhgg.2021.100026"},{"key":"2025090317402788000_2025.08.30.673262v1.10","doi-asserted-by":"crossref","unstructured":"A. Nair , A. Greeny , R. Rajendran , M. A. Abdelgawad , M. M. Ghoneim , R. P. Raghavan , S. T. Sudevan , B. Mathew , H. Kim , KIF1A-Associated Neurological Disorder: An Overview of a Rare Mutational Disease. Pharmaceuticals (Basel) 16, (2023).","DOI":"10.3390\/ph16020147"},{"key":"2025090317402788000_2025.08.30.673262v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-21509-5"},{"key":"2025090317402788000_2025.08.30.673262v1.12","doi-asserted-by":"crossref","unstructured":"B. G. Budaitis , S. Jariwala , L. Rao , Y. Yue , D. Sept , K. J. Verhey , A. Gennerich , Pathogenic mutations in the kinesin-3 motor KIF1A diminish force generation and movement through allosteric mechanisms. Journal of Cell Biology 220, (2021).","DOI":"10.1083\/jcb.202004227"},{"key":"2025090317402788000_2025.08.30.673262v1.13","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abf1002"},{"key":"2025090317402788000_2025.08.30.673262v1.14","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2113795119"},{"key":"2025090317402788000_2025.08.30.673262v1.15","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-48720-4"},{"key":"2025090317402788000_2025.08.30.673262v1.16","doi-asserted-by":"crossref","first-page":"656","DOI":"10.3390\/biom15050656","article-title":"Distinct Clinical Phenotypes in KIF1A-Associated Neurological Disorders Result from Different Amino Acid Substitutions at the Same Residue in KIF1A","volume":"15","year":"2025","journal-title":"Biomolecules"},{"key":"2025090317402788000_2025.08.30.673262v1.17","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1905690116"},{"key":"2025090317402788000_2025.08.30.673262v1.18","doi-asserted-by":"crossref","unstructured":"K. Chiba , T. Kita , Y. Anazawa , S. Niwa , Insight into the regulation of axonal transport from the study of KIF1A-associated neurological disorder. Journal of Cell Science 136, (2023).","DOI":"10.1242\/jcs.260742"},{"key":"2025090317402788000_2025.08.30.673262v1.19","doi-asserted-by":"publisher","DOI":"10.1038\/ejhg.2011.261"},{"key":"2025090317402788000_2025.08.30.673262v1.20","doi-asserted-by":"publisher","DOI":"10.1042\/BST20221238"},{"key":"2025090317402788000_2025.08.30.673262v1.21","doi-asserted-by":"crossref","unstructured":"C. Aguilera , S. H\u00fcmmer , M. Masanas , E. Gabau , M. Guitart , A. A. Jeyaprakash , M. F. Segura , A. Santamaria , A. Ruiz , The Novel KIF1A Missense Variant (R169T) Strongly Reduces Microtubule Stimulated ATPase Activity and Is Associated With NESCAV Syndrome. Frontiers in Neuroscience 15, (2021).","DOI":"10.3389\/fnins.2021.618098"},{"key":"2025090317402788000_2025.08.30.673262v1.22","doi-asserted-by":"crossref","first-page":"5057","DOI":"10.1007\/s00415-023-11816-w","article-title":"A new genetic cause of spastic ataxia: the p.Glu415Lys variant in TUBA4A","volume":"270","year":"2023","journal-title":"J. Neurol"},{"key":"2025090317402788000_2025.08.30.673262v1.23","doi-asserted-by":"publisher","DOI":"10.1002\/humu.22709"},{"key":"2025090317402788000_2025.08.30.673262v1.24","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-32048-y"},{"key":"2025090317402788000_2025.08.30.673262v1.25","doi-asserted-by":"publisher","DOI":"10.1038\/S41586-024-07487-W"},{"key":"2025090317402788000_2025.08.30.673262v1.26","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.1000072"},{"key":"2025090317402788000_2025.08.30.673262v1.27","doi-asserted-by":"crossref","unstructured":"Terrific Broth. Cold Spring Harbor Protocols 2006, pdb.rec8620 (2006).","DOI":"10.1101\/pdb.rec8620"},{"key":"2025090317402788000_2025.08.30.673262v1.28","doi-asserted-by":"publisher","DOI":"10.1107\/S2052252522010624"},{"key":"2025090317402788000_2025.08.30.673262v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2005.03.010"},{"key":"2025090317402788000_2025.08.30.673262v1.30","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1038\/nmeth.4169","article-title":"cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination","volume":"14","year":"2017","journal-title":"Nature Methods"},{"key":"2025090317402788000_2025.08.30.673262v1.31","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-23581-3"},{"key":"2025090317402788000_2025.08.30.673262v1.32","doi-asserted-by":"publisher","DOI":"10.1002\/pro.3235"},{"key":"2025090317402788000_2025.08.30.673262v1.33","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.20084"},{"key":"2025090317402788000_2025.08.30.673262v1.34","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444904019158"},{"key":"2025090317402788000_2025.08.30.673262v1.35","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2009.07.008"},{"key":"2025090317402788000_2025.08.30.673262v1.36","doi-asserted-by":"publisher","DOI":"10.1002\/pro.3330"},{"key":"2025090317402788000_2025.08.30.673262v1.37","doi-asserted-by":"publisher","DOI":"10.1107\/S2059798318006551"},{"key":"2025090317402788000_2025.08.30.673262v1.38","doi-asserted-by":"publisher","DOI":"10.1107\/S2059798318002425"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.08.30.673262","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T09:27:32Z","timestamp":1768469252000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.08.30.673262"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,30]]},"references-count":38,"URL":"https:\/\/doi.org\/10.1101\/2025.08.30.673262","relation":{},"subject":[],"published":{"date-parts":[[2025,8,30]]},"subtype":"preprint"}}