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PreprintCited by 9Alzheimer's disease research and treatmentsMicrotubule and mitosis dynamicsNeuroinflammation and Neurodegeneration Mechanisms

A pathological phosphorylation pattern enhances tau cooperativity on microtubules and facilitates tau filament assembly

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Fan, Xiangyu, Okada, Kyoko, Lin, Henry, Ori-McKenney, Kassandra M., McKenney, Richard J.·2025·Communications Biology
DOI 10.1101/2025.01.29.635117PMC PMC13714281
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Abstract

Phosphorylation plays a crucial role in both normal and disease processes involving the microtubule-associated protein tau. Physiologically, phosphorylation regulates tau’s subcellular localization within neurons and is involved in fetal development and animal hibernation. However, abnormal phosphorylation of tau is linked to the formation of neurofibrillary tangles (NFTs) in various human tauopathies. Interestingly, the patterns of tau phosphorylation are similar in both normal and abnormal processes, leaving unclear whether phosphorylated tau retains its functional role in normal processes. The relationship between tau phosphorylation and NFT assembly in tauopathies is also still debated. To address these questions, we investigated the effects of tau phosphorylation on microtubule binding, cooperative protein envelope formation, and NFT filament assembly relevant to tauopathies. Consistent with previous results, our findings show that tau phosphorylation decreases tau’s overall affinity for microtubules, but we reveal that phosphorylation more dramatically impacts the cooperativity between tau molecules during tau envelope formation along microtubules. Additionally, we observed that the specific pattern of phosphorylation, rather than overall phosphorylation level, strongly impacts the assembly of tau filaments in vitro . Our results reveal new insights into how tau phosphorylation impacts tau’s physiological roles on microtubules and its pathoconversion into NFTs.

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  • Tau expressed in HEK cells is phosphorylated by endogenous kinases at 58 residues, with <15% occupancy, indicating substantial heterogeneity among tau molecules.

    “For example, tau expressed in HEK cells is phosphorylated by endogenous kinases at 58 residues, with <15% occupancy, indicating substantial heterogeneity among tau molecules (Fan et al., 2025).”

    From Tau hyperphosphorylation impairs cooperative binding to microtubules and perturbs organelle trafficking in neurons (2026) · cites this paper as Fan et al., 2025

    The Characterization section (p. 4) reports 52 phosphorylated sites in HEK tau, not 58; 58 is the count for Sf9 tau. The same section attributes phosphorylation in these cells to endogenous kinases. On p. 5, the paper says occupancy is below approximately 15% at most sites, not all sites, which is consistent with variation among tau molecules. (LLM, conf=0.92)

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  1. [1]1. Weingarten, M.D., Lockwood, A. H., Hwo, S. Y ., &amp; Kirschner, M. W. A protein factor essential for microtubule assembly. Proceedings of the National Academy of Sciences of the United States of America 72, 1858-1862 (1975).
  2. [2]2. Cleveland, D.W., Hwo, S. Y ., &amp; Kirschner, M. W. . Puri2cation of tau, a microtubuleassociated protein that induces assembly of microtubules from puri2ed tubulin. Journal of molecular biology 116, 207-225 (1977).
  3. [5]5. Kimura, T. et al. Microtubule-associated protein tau is essential for long-term depression in the hippocampus. Philosophical Transactions of the Royal Society B: Biological Sciences 369(2014).
  4. [9]9. Violet, M. et al. A major role for Tau in neuronal DNA and RNA protection in vivo under physiological and hyperthermic conditions. Front Cell Neurosci 8, 84 (2014).
  5. [12]12. Portillo, M. et al. SIRT6-CBP-dependent nuclear Tau accumulation and its role in protein synthesis. Cell Rep 35, 109035 (2021).
  6. [11]11. Sotiropoulos, I. et al. Atypical, non-standard functions of the microtubule associated Tau protein. Acta Neuropathol Commun 5, 91 (2017).
  7. [16]16. Beaudet, D., Berger, C.L. &amp; Hendricks, A.G. The types and numbers of kinesins and dyneins transporting endocytic cargoes modulate their motility and response to tau. J Biol Chem 300, 107323 (2024).
  8. [21]21. Kosik, K.S., Joachim, C. L., &amp; Selkoe, D. J. Microtubule-associated protein tau (tau) is a major antigenic component of paired helical 2laments in Alzheimer disease. Proceedings of the National Academy of Sciences of the United States of America 83, 4044-4048 (1986).
  9. [20]20. Grundke-Iqbal, I., Iqbal, K., Tung, Y . C., Quinlan, M., Wisniewski, H. M., &amp; Binder, L. I. Abnormal phosphorylation of the microtubule-associated protein tau (tau) in Alzheimer cytoskeletal pathology. Proceedings of the National Academy of Sciences of the United States of America 83, 4913-4917 (1986).
  10. [22]22. Wood, J.G., Mirra, S. S., Pollock, N. J., &amp; Binder, L. I. Neuro2brillary tangles of Alzheimer disease share antigenic determinants with the axonal microtubule-associated protein tau (tau). Proceedings of the National Academy of Sciences of the United States of America 83, 4040-4043 (1986).
  11. [25]25. Pollock, N.J., Mirra, S. S., Binder, L. I., Hansen, L. A., &amp; Wood, J. G. Filamentous aggregates in Pick's disease, progressive supranuclear palsy, and Alzheimer's disease share antigenic determinants with microtubule-associated protein, tau. Lancet (London, England) 2, 1211 (1986).
  12. [30]30. Drewes, G., Trinczek, B., Illenberger, S., Biernat, J., Schmitt-Ulms, G., Meyer, H. E., Mandelkow, E. M., &amp; Mandelkow, E. Microtubule-associated protein/microtubule af2nityregulating kinase (p110mark). A novel protein kinase that regulates tau-microtubule interactions and dynamic instability by phosphorylation at the Alzheimer-speci2c site serine 262. The Journal of biological chemistry 270, 7679-7688 (1995).
  13. [29]29. Zhang, W. et al. Novel tau 2lament fold in corticobasal degeneration. Nature 580, 283287 (2020).
  14. [36]36. Despres, C. et al. Identi2cation of the Tau phosphorylation pattern that drives its aggregation. Proceedings of the National Academy of Sciences 114, 9080-9085 (2017).
  15. [37]37. Jeganathan, S. et al. Proline-directed Pseudo-phosphorylation at AT8 and PHF1 Epitopes Induces a Compaction of the Paperclip Folding of Tau and Generates a Pathological (MC1) Conformation. Journal of Biological Chemistry 283, 32066-32076 (2008).
  16. [42]42. Su, B. et al. Physiological regulation of tau phosphorylation during hibernation. Journal of Neurochemistry 105, 2098-2108 (2008).
  17. [41]41. Arendt, T ., Stieler, J., Strijkstra, A. M., Hut, R. A., Rüdiger, J., Van der Zee, E. A., Harkany, T ., Holzer, M., &amp; Härtig, W. Reversible paired helical 2lament-like phosphorylation of tau is an adaptive process associated with neuronal plasticity in hibernating animals. The Journal of neuroscience : the of2cial journal of the Society for Neuroscience 23, 6972-6981 (2003).
  18. [44]44. Brion, J.P ., Smith, C., Couck, A. M., Gallo, J. M., &amp; Anderton, B. H. Developmental changes in tau phosphorylation: fetal tau is transiently phosphorylated in a manner similar to paired helical 2lament-tau characteristic of Alzheimer's disease. Journal of neurochemistry 61, 2071-2080 (1993).
  19. [48]48. Yu, Y. et al. Developmental regulation of tau phosphorylation, tau kinases, and tau phosphatases. Journal of Neurochemistry 108, 1480-1494 (2009).
  20. [53]53. Mietelska-Porowska, A., Wasik, U., Goras, M., Filipek, A. &amp; Niewiadomska, G. T au Protein Modi2cations and Interactions: Their Role in Function and Dysfunction. International Journal of Molecular Sciences 15, 4671-4713 (2014).
  21. [56]56. Siahaan, V. et al. T au phosphorylation impedes functionality of protective tau envelopes. BioRxiv (2024).
  22. [60]60. Hasegawa, M., Morishima-Kawashima, M., Takio, K., Suzuki, M., Titani, K., &amp; Ihara, Y . Protein sequence and mass spectrometric analyses of tau in the Alzheimer's disease brain. The Journal of biological chemistry 267, 17047-17054 (1992).
  23. [62]62. Hanger, D.P ., Betts, J. C., Loviny, T. L., Blackstock, W. P ., &amp; Anderton, B. H. New phosphorylation sites identi2ed in hyperphosphorylated tau (paired helical 2lament-tau) from Alzheimer's disease brain using nanoelectrospray mass spectrometry. Journal of neurochemistry 71, 2465-2476 (1998).
  24. [61]61. Morishima-Kawashima, M., Hasegawa, M., Takio, K., Suzuki, M., Yoshida, H., Titani, K., &amp; Ihara, Y. Proline-directed and non-proline-directed phosphorylation of PHF-tau. The Journal of biological chemistry 207, 823-829 (1995).
  25. [63]63. Hanger, D.P. et al. Novel Phosphorylation Sites in Tau from Alzheimer Brain Support a Role for Casein Kinase 1 in Disease Pathogenesis. Journal of Biological Chemistry 282, 23645-23654 (2007).
  26. [66]66. Köpke, E., Tung, Y . C., Shaikh, S., Alonso, A. C., Iqbal, K., &amp; Grundke-Iqbal, I. Microtubuleassociated protein tau. Abnormal phosphorylation of a non-paired helical 2lament pool in Alzheimer disease. The Journal of biological chemistry 268, 24374-24384 (1993).
  27. [69]69. Cho, J.H., &amp; Johnson, G. V. Primed phosphorylation of tau at Thr231 by glycogen synthase kinase 3beta (GSK3beta) plays a critical role in regulating tau's ability to bind and stabilize microtubules. Journal of neurochemistry 88, 349-358 (2004).
  28. [71]71. Tepper, K. et al. Oligomer Formation of Tau Protein Hyperphosphorylated in Cells. Journal of Biological Chemistry 289, 34389-34407 (2014).
  29. [75]75. Pollack, S.J. et al. Paired Helical Filament-Forming Region of Tau (297-391) InVuences Endogenous Tau Protein and Accumulates in Acidic Compartments in Human Neuronal Cells. Journal of Molecular Biology 432, 4891-4907 (2020).
  30. [76]76. Kaniyappan, S. et al. FRET-based Tau seeding assay does not represent prion-like templated assembly of Tau 2laments. Mol Neurodegener 15, 39 (2020).
  31. [78]78. Run, X. et al. Anesthesia Induces Phosphorylation of T au. Journal of Alzheimer's Disease 16, 619-626 (2009).
  32. [79]79. Planel, E. et al. Anesthesia Leads to Tau Hyperphosphorylation through Inhibition of Phosphatase Activity by Hypothermia. The Journal of Neuroscience 27, 3090-3097 (2007).
  33. [80]80. Alonso , A., Zaidi, T ., Novak, M., Grundke-Iqbal, I., &amp; Iqbal, K. Hyperphosphorylation induces self-assembly of tau into tangles of paired helical 2laments/straight 2laments. Proceedings of the National Academy of Sciences of the United States of America 98, 6923-6928 (2001).
  34. [87]87. Gyparaki, M.T. et al. Tau forms oligomeric complexes on microtubules that are distinct from tau aggregates. Proceedings of the National Academy of Sciences 118(2021).
  35. [86]86. Dickey, C.A. et al. The high-af2nity HSP90-CHIP complex recognizes and selectively degrades phosphorylated tau client proteins. Journal of Clinical Investigation 117, 648658 (2007).
  36. [88]88. Haj-Yahya, M. et al. Site-Speci2c Hyperphosphorylation Inhibits, Rather than Promotes, Tau Fibrillization, Seeding Capacity, and Its Microtubule Binding. Angewandte Chemie International Edition 59, 4059-4067 (2020).
  37. [90]90. Wegmann, S. et al. Tau protein liquid-liquid phase separation can initiate tau aggregation. The EMBO Journal 37(2018).
  38. [89]89. Ambadipudi, S., Biernat, J., Riedel, D., Mandelkow, E. &amp; Zweckstetter, M. Liquid-liquid phase separation of the microtubule-binding repeats of the Alzheimer-related protein Tau. Nat Commun 8, 275 (2017).
  39. [91]91. Kanaan, N.M., Hamel, C., Grabinski, T . &amp; Combs, B. Liquid-liquid phase separation induces pathogenic tau conformations in vitro. Nat Commun 11, 2809 (2020).
  40. [93]93. Monroy, B.Y. et al. Competition between microtubule-associated proteins directs motor transport. Nat Commun 9, 1487 (2018).
  41. [92]92. Hochmair, J. et al. Molecular crowding and RNA synergize to promote phase separation, microtubule interaction, and seeding of T au condensates. EMBO J 41, e108882 (2022).

Cited by

Sorted by citations

Authors
Fan, X., Okada, K., Lin, H., Ori-McKenney, K. M., & McKenney, R. J.
Affiliations
  • Henan University (CN)
  • University of California, Davis (US)
Corresponding author
McKenney, Richard J.
ORCID
Venue
Communications Biology
Publisher
openRxiv
Published
2025-01-31
Volume
9
Issue
1
ISSN
2399-3642 (ISSN-L)
DOI
10.1101/2025.01.29.635117
PMC
PMC13714281
OpenAlex
W4407021530
Cited by
9
References
0
Citations by year
2026: 8 · 2025: 1
FWCI
36.09
Type
journal article
Language
en
Open access
Licence
Locations
  • Communications Biology (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc)
  • Research Square (acceptedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by-nc)
  • PubMed Central (submittedVersion, cc-by)
Funders
  • Alzheimer's Association (23AARGD-1022898)
  • American Heart Association (25PRE1362890)
  • University of California, Davis
  • National Institute on Aging (P30AG072972)
Topics
  • Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
Keywords
tau phosphorylation, microtubule binding, neurofibrillary tangles, Alzheimer's disease, microtubule-associated proteins

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