{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T04:35:52Z","timestamp":1787978152324,"version":"build-2784847793"},"reference-count":51,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,2,26]],"date-time":"2025-02-26T00:00:00Z","timestamp":1740528000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NINDS","award":["Q-2097-20220331"],"award-info":[{"award-number":["Q-2097-20220331"]}]},{"name":"NINDS","award":["R01 NS105874"],"award-info":[{"award-number":["R01 NS105874"]}]},{"name":"NINDS","award":["R01 GM122763"],"award-info":[{"award-number":["R01 GM122763"]}]},{"DOI":"10.13039\/100000928","name":"Welch Foundation","doi-asserted-by":"publisher","award":["Q-2097-20220331"],"award-info":[{"award-number":["Q-2097-20220331"]}],"id":[{"id":"10.13039\/100000928","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000928","name":"Welch Foundation","doi-asserted-by":"publisher","award":["R01 NS105874"],"award-info":[{"award-number":["R01 NS105874"]}],"id":[{"id":"10.13039\/100000928","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000928","name":"Welch Foundation","doi-asserted-by":"publisher","award":["R01 GM122763"],"award-info":[{"award-number":["R01 GM122763"]}],"id":[{"id":"10.13039\/100000928","id-type":"DOI","asserted-by":"publisher"}]},{"name":"NIGMS","award":["Q-2097-20220331"],"award-info":[{"award-number":["Q-2097-20220331"]}]},{"name":"NIGMS","award":["R01 NS105874"],"award-info":[{"award-number":["R01 NS105874"]}]},{"name":"NIGMS","award":["R01 GM122763"],"award-info":[{"award-number":["R01 GM122763"]}]},{"name":"NIH","award":["Q-2097-20220331"],"award-info":[{"award-number":["Q-2097-20220331"]}]},{"name":"NIH","award":["R01 NS105874"],"award-info":[{"award-number":["R01 NS105874"]}]},{"name":"NIH","award":["R01 GM122763"],"award-info":[{"award-number":["R01 GM122763"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Tau is a microtubule-associated protein that undergoes liquid\u2013liquid phase separation (LLPS) to form condensates under physiological conditions, facilitating microtubule stabilization and intracellular transport. LLPS has also been implicated in pathological Tau aggregation, which contributes to tauopathies such as Alzheimer\u2019s disease. While LLPS is known to promote Tau aggregation, the relationship between Tau\u2019s structural states and its phase separation behavior remains poorly defined. Here, we examine how oligomerization modulates Tau LLPS and uncover key distinctions between monomeric, oligomeric, and amyloidogenic Tau species. Using dynamic light scattering and fluorescence microscopy, we monitored oligomer formation over time and assessed oligomeric Tau\u2019s ability to undergo LLPS. We found that Tau monomers readily phase separate and form condensates. As oligomerization progresses, Tau\u2019s propensity to undergo LLPS diminishes, with oligomers still being able to phase separate, albeit with reduced efficiency. Interestingly, oligomeric Tau is recruited into condensates formed with 0-day-aged Tau, with this recruitment depending on the oligomer state of maturation. Early-stage, Thioflavin T (ThT)-negative oligomers co-localize with 0-day-aged Tau condensates, whereas ThT-positive oligomers resist condensate recruitment entirely. This study highlights a dynamic interplay between Tau LLPS and aggregation, providing insight into how Tau\u2019s structural and oligomeric states influence its pathological and functional roles. These findings underscore the need to further explore LLPS as a likely modulator of Tau pathogenesis and distinct pathogenic oligomers as viable therapeutic targets in tauopathies.<\/jats:p>","DOI":"10.3390\/biom15030336","type":"journal-article","created":{"date-parts":[[2025,2,26]],"date-time":"2025-02-26T06:15:33Z","timestamp":1740550533000},"page":"336","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Tau Oligomers Resist Phase Separation"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8493-0015","authenticated-orcid":false,"given":"Lathan","family":"Lucas","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, TX 77030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8160-5607","authenticated-orcid":false,"given":"Phoebe S.","family":"Tsoi","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, TX 77030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3175-5700","authenticated-orcid":false,"given":"Josephine C.","family":"Ferreon","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, TX 77030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8538-1732","authenticated-orcid":false,"given":"Allan Chris M.","family":"Ferreon","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, TX 77030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1152\/physrev.1995.75.4.835","article-title":"Role of microtubule-associated proteins in the control of microtubule assembly","volume":"75","author":"Maccioni","year":"1995","journal-title":"Physiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Dehmelt, L., and Halpain, S. (2005). The MAP2\/Tau family of microtubule-associated proteins. Genome Biol., 6.","DOI":"10.1186\/gb-2004-6-1-204"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Barbier, P., Zejneli, O., Martinho, M., Lasorsa, A., Belle, V., Smet-Nocca, C., Tsvetkov, P.O., Devred, F., and Landrieu, I. (2019). Role of tau as a microtubule-associated protein: Structural and functional aspects. Front. Aging Neurosci., 11.","DOI":"10.3389\/fnagi.2019.00204"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.pneurobio.2008.03.002","article-title":"Microtubule-associated protein tau in development, degeneration and protection of neurons","volume":"85","author":"Wang","year":"2008","journal-title":"Prog. Neurobiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1038\/nrn.2015.1","article-title":"Tau in physiology and pathology","volume":"17","author":"Wang","year":"2016","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1007\/s00401-017-1707-9","article-title":"Roles of tau protein in health and disease","volume":"133","author":"Guo","year":"2017","journal-title":"Acta Neuropathol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1038\/s41580-024-00753-9","article-title":"Cellular and pathological functions of tau","volume":"25","author":"Naguib","year":"2024","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.bbadis.2004.09.008","article-title":"Tau pathology in Alzheimer disease and other tauopathies","volume":"1739","author":"Iqbal","year":"2005","journal-title":"Biochim. Biophys. Acta (BBA)-Mol. Basis Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"656","DOI":"10.2174\/156720510793611592","article-title":"Tau in Alzheimer disease and related tauopathies","volume":"7","author":"Iqbal","year":"2010","journal-title":"Curr. Alzheimer Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1007\/s10571-017-0574-1","article-title":"Tau proteins and tauopathies in Alzheimer\u2019s disease","volume":"38","author":"Chong","year":"2018","journal-title":"Cell. Mol. Neurobiol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Tsoi, P.S., Quan, M.D., Ferreon, J.C., and Ferreon, A.C.M. (2023). Aggregation of disordered proteins associated with neurodegeneration. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24043380"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"591","DOI":"10.2174\/156720508786898442","article-title":"Tau oligomerization: A role for tau aggregation intermediates linked to neurodegeneration","volume":"5","author":"Sahara","year":"2008","journal-title":"Curr. Alzheimer Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.1111\/j.1471-4159.2009.06511.x","article-title":"Tau oligomers and aggregation in Alzheimer\u2019s disease","volume":"112","year":"2010","journal-title":"J. Neurochem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.1021\/cn500148z","article-title":"The role of tau oligomers in the onset of Alzheimer\u2019s disease neuropathology","volume":"5","author":"DeRosa","year":"2014","journal-title":"ACS Chem. Neurosci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3856","DOI":"10.1021\/bi061359o","article-title":"Granular tau oligomers as intermediates of tau filaments","volume":"46","author":"Maeda","year":"2007","journal-title":"Biochemistry"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"33298","DOI":"10.1074\/jbc.M305207200","article-title":"Microtubule-dependent oligomerization of tau: Implications for physiological tau function and tauopathies","volume":"278","author":"Makrides","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1111\/jnc.13604","article-title":"Oligomerization of the microtubule-associated protein tau is mediated by its N-terminal sequences: Implications for normal and pathological tau action","volume":"137","author":"Feinstein","year":"2016","journal-title":"J. Neurochem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e2021461118","DOI":"10.1073\/pnas.2021461118","article-title":"Tau forms oligomeric complexes on microtubules that are distinct from tau aggregates","volume":"118","author":"Gyparaki","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Iqbal, K., del C. Alonso, A., Gong, C.X., Khatoon, S., Pei, J.J., Wang, J.Z., and Grundke-Iqbal, I. (1998). Mechanisms of Neurofibrillary Degeneration and the Formation of Neurofibrillary Tangles, Springer.","DOI":"10.1007\/978-3-7091-6467-9_15"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1774","DOI":"10.1529\/biophysj.105.070136","article-title":"Structure of core domain of fibril-forming PHF\/Tau fragments","volume":"90","author":"Inouye","year":"2006","journal-title":"Biophys. J."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1083","DOI":"10.1007\/s10571-006-9083-3","article-title":"Intrinsically disordered proteins in the neurodegenerative processes: Formation of tau protein paired helical filaments and their analysis","volume":"26","author":"Skrabana","year":"2006","journal-title":"Cell. Mol. Neurobiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e98049","DOI":"10.15252\/embj.201798049","article-title":"Tau protein liquid\u2013liquid phase separation can initiate tau aggregation","volume":"37","author":"Wegmann","year":"2018","journal-title":"EMBO J."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zhang, X., Lin, Y., Eschmann, N.A., Zhou, H., Rauch, J.N., Hernandez, I., Guzman, E., Kosik, K.S., and Han, S. (2017). RNA stores tau reversibly in complex coacervates. PLoS Biol., 15.","DOI":"10.1101\/111245"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2304","DOI":"10.1016\/j.celrep.2017.08.042","article-title":"Local nucleation of microtubule bundles through tubulin concentration into a condensed Tau phase","volume":"20","author":"Braun","year":"2017","journal-title":"Cell Rep."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1038\/s41467-017-00480-0","article-title":"Liquid-liquid phase separation of the microtubule-binding re- peats of the Alzheimer-related protein Tau","volume":"8","author":"Ambadipudi","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Ferreon, J.C., Jain, A., Choi, K.J., Tsoi, P.S., MacKenzie, K.R., Jung, S.Y., and Ferreon, A.C. (2018). Acetylation disfavors tau phase separation. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19051360"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e42571","DOI":"10.7554\/eLife.42571","article-title":"Narrow equilibrium window for complex coacervation of tau and RNA under cellular conditions","volume":"8","author":"Lin","year":"2018","journal-title":"eLife"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1007\/s12551-020-00680-x","article-title":"Biological phase separation: Cell biology meets biophysics","volume":"12","author":"Yoshizawa","year":"2020","journal-title":"Biophys. Rev."},{"key":"ref_29","first-page":"3766","article-title":"Liquid-liquid phase separation (LLPS) in cellular physiology and tumor biology","volume":"11","author":"Peng","year":"2021","journal-title":"Am. J. Cancer Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1016\/j.tips.2021.07.003","article-title":"Liquid\u2013liquid phase separation: A principal organizer of the cell\u2019s biochemical activity architecture","volume":"42","author":"Zhang","year":"2021","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Nesterov, S.V., Ilyinsky, N.S., and Uversky, V.N. (2021). Liquid-liquid phase separation as a common organizing principle of intracellular space and biomembranes providing dynamic adaptive responses. Biochim. Biophys. Acta (BBA)-Mol. Cell Res., 1868.","DOI":"10.1016\/j.bbamcr.2021.119102"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5579","DOI":"10.1038\/s41467-021-25761-7","article-title":"Liquid condensation of reprogramming factor KLF4 with DNA provides a mechanism for chromatin organization","volume":"12","author":"Sharma","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"737","DOI":"10.1038\/s41556-022-00896-x","article-title":"NANOG prion-like assembly mediates DNA bridging to facilitate chromatin reorganization and activation of pluripotency","volume":"24","author":"Choi","year":"2022","journal-title":"Nat. Cell Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1078","DOI":"10.1038\/s41556-019-0375-5","article-title":"Microtubules gate tau condensation to spatially regulate microtubule functions","volume":"21","author":"Tan","year":"2019","journal-title":"Nat. Cell Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/j.semcdb.2019.06.007","article-title":"Biomolecular condensation of the microtubule-associated protein tau","volume":"99","author":"Wegmann","year":"2020","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e108882","DOI":"10.15252\/embj.2021108882","article-title":"Molecular crowding and RNA synergize to promote phase separation, microtubule interaction, and seeding of Tau condensates","volume":"41","author":"Hochmair","year":"2022","journal-title":"EMBO J."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"31882","DOI":"10.1073\/pnas.2012460117","article-title":"Regulatory mechanisms of tau protein fibrillation under the conditions of liquid\u2013liquid phase separation","volume":"117","author":"Boyko","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Lin, Y., Fichou, Y., Longhini, A.P., Llanes, L.C., Yin, P., Bazan, G.C., Kosik, K.S., and Han, S. (2021). Liquid-liquid phase separation of tau driven by hydrophobic interaction facilitates fibrillization of tau. J. Mol. Biol., 433.","DOI":"10.1016\/j.jmb.2020.166731"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1016\/j.tcb.2022.01.011","article-title":"Tau liquid\u2013liquid phase separation in neurodegenerative diseases","volume":"32","author":"Boyko","year":"2022","journal-title":"Trends Cell Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1694","DOI":"10.1021\/bi0357006","article-title":"Tau paired helical filaments from Alzheimer\u2019s disease brain and assembled in vitro are based on \u03b2-structure in the core domain","volume":"43","author":"Barghorn","year":"2004","journal-title":"Biochemistry"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1948","DOI":"10.1021\/bi00859a010","article-title":"Spectroscopic determination of tryptophan and tyrosine in proteins","volume":"6","author":"Edelhoch","year":"1967","journal-title":"Biochemistry"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/0003-2697(89)90602-7","article-title":"Calculation of protein extinction coefficients from amino acid sequence data","volume":"182","author":"Gill","year":"1989","journal-title":"Anal. Biochem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"13357","DOI":"10.1021\/bi048666j","article-title":"Thermodynamics of denaturant-induced unfolding of a protein that exhibits variable two-state denaturation","volume":"43","author":"Ferreon","year":"2004","journal-title":"Biochemistry"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1002\/cm.21772","article-title":"Big tau: What, how, where and why","volume":"81","author":"Fischer","year":"2024","journal-title":"Cytoskeleton"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"20318","DOI":"10.1074\/jbc.M114.554725","article-title":"Stages and conformations of the Tau repeat domain during aggregation and its effect on neuronal toxicity","volume":"289","author":"Kumar","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"4231","DOI":"10.1038\/s41467-021-24362-8","article-title":"Co-factor-free aggregation of tau into seeding-competent RNA-sequestering amyloid fibrils","volume":"12","author":"Chakraborty","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1999","DOI":"10.2147\/IJN.S494104","article-title":"Temperature-Dependent Aggregation of Tau Protein Is Attenuated by Native PLGA Nanoparticles Under in vitro Conditions","volume":"20","author":"Paul","year":"2025","journal-title":"Int. J. Nanomed."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"11054","DOI":"10.1074\/jbc.AC119.009198","article-title":"Liquid\u2013liquid phase separation of tau protein: The crucial role of electrostatic interactions","volume":"294","author":"Boyko","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"7445","DOI":"10.1073\/pnas.0802036105","article-title":"Complementary dimerization of microtubule-associated tau protein: Implications for microtubule bundling and tau-mediated pathogenesis","volume":"105","author":"Rosenberg","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Castro, T.G., Munteanu, F.D., and Cavaco-Paulo, A. (2019). Electrostatics of tau protein by molecular dynamics. Biomolecules, 9.","DOI":"10.3390\/biom9030116"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"6822","DOI":"10.1021\/acs.biochem.8b01051","article-title":"A chemical chaperone decouples TDP-43 disordered domain phase separation from fibrillation","volume":"57","author":"Choi","year":"2018","journal-title":"Biochemistry"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/15\/3\/336\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:42:40Z","timestamp":1760028160000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/15\/3\/336"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,26]]},"references-count":51,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["biom15030336"],"URL":"https:\/\/doi.org\/10.3390\/biom15030336","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,26]]}}}