Graded regulation of microtubule-binding of Tau by the phosphorylation state of the proline-rich region in living neurons
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Abstract
Tau protein is a microtubule-associated protein that plays a crucial role in maintaining neuronal morphology and axonal transport. While phosphorylation is known to regulate Tau-microtubule interactions, the contribution of specific phosphorylation patterns in situ remains poorly understood due to the complexity of the intracellular environment. In this study, we combined fluorescence recovery after photobleaching (FRAP) in primary cultured rat hippocampal neurons with dephosphorylation-mimetic mutations and computational modeling to analyze the effects of phosphorylation on Tau-microtubule interaction. We particularly focused on the proline-rich region, of which phosphorylation has been studied in physiological and pathological perspectives, and generated a dephosphorylation-mimetic Tau mutant by substituting key phosphorylation sites with alanine residues and compared its microtubule-binding dynamics to those of WT-Tau in FRAP experiments. Experimental data, together with simulation-based parameter exploration, revealed that the overall number of non-phosphorylated sites plays a more dominant role than their specific locations in modulating Tau-microtubule affinity. These findings provide new insights into the post-translational regulation of Tau and establish a computational-experimental framework for interrogating intracellular protein dynamics.
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Our recent study suggests that the total number of non-phosphorylated sites in the PRR is more critical for stronger MT-binding of Tau than the status of specific sites in the PRR.
“Furthermore, our recent study suggests that the total number of non-phosphorylated sites in the PRR is more critical for stronger MT-binding of Tau than the status of specific sites in the PRR (43).”
From Active transport by cytoplasmic dynein sustains dendritic MAP2 localization in developing neurons (2026) · cites this paper as [43]
The section ‘The Number of Dephospho-mimetic Sites in PRR2 Gradually Modulates Tau-MT Binding’ says that the number of non-phosphorylated residues matters more than the status of a few critical sites. The study tested dephospho-mimetic sites in PRR2, however, so it does not establish the claim for the broader proline-rich region (PRR). (LLM, conf=0.82)
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Active transport by cytoplasmic dynein sustains dendritic MAP2 localization in developing neurons
Yonemura, Y., Sakai, Y., Nakata, R., Hagita-Tatsumoto, A., Miyasaka, T., & Misonou, H. (2026) · Journal of Biological Chemistry · doi:10.1016/j.jbc.2026.113407
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Nimavard, R. T., Sadeghi, S. A., Colón-Rosado, J. A., Yue, Y., Fang, F., Hagar, H. C., Kuo, M.-H., & Sun, L. (2026) · Journal of Proteome Research · doi:10.1021/acs.jproteome.6c00539
- Authors
- Nakata, R., Torii, T., Miyasaka, T., & Misonou, H.
- Affiliations
- Doshisha University (JP)
- Kitasato University (JP)
- Nihon University (JP)
- Corresponding author
- Misonou, Hiroaki
- Venue
- Neuroscience Research
- Publisher
- Elsevier BV
- Published
- 2026-01
- Volume
- 222
- Pages
- 104987
- ISSN
- 0168-0102 (ISSN-L)
- OpenAlex
- W4416416062
- Cited by
- 3
- References
- 53
- Citations by year
- 2026: 3
- FWCI
- 1.94
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Neuroscience Research (publishedVersion, cc-by-nc-nd)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed (publishedVersion)
- Funders
- Japan Science and Technology Agency (JPMJSP2129)
- Japan Society for the Promotion of Science (23K24207)
- National Institute of General Medical Sciences
- Japan Science Society
- National Institutes of Health (R24 GM137787)
- 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 protein, microtubule binding, proline-rich region, phosphorylation, hippocampal neurons, axonal transport, neuronal morphology, post-translational regulation, protein dynamics, computational modeling
- MeSH terms
- Animals, Cells, Cultured, Hippocampus, Microtubules, Neurons, Phosphorylation, Proline, Protein Binding, tau Proteins, Fluorescence Recovery After Photobleaching, Rats
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