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Mechanisms of Regulation and Diverse Activities of Tau-Tubulin Kinase (TTBK) Isoforms

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Abstract

Tau-tubulin kinase 1 (TTBK1) is a CNS-specific, kinase that has been implicated in the pathological phosphorylation of tau in Alzheimer’s Disease (AD) and Frontotemporal Dementia (FTD). TTBK1 is a challenging therapeutic target because it shares a highly conserved catalytic domain with its homolog, TTBK2, a ubiquitously expressed kinase genetically linked to the disease spinocerebellar ataxia type 11. The present study attempts to elucidate the functional distinctions between the TTBK isoforms and increase our understanding of them as distinct targets for the treatment of neurodegenerative disease. We demonstrate that in cortical neurons, TTBK1, not TTBK2, is the isoform responsible for tau phosphorylation at epitopes enriched in tauopathies such as Serine 422. In addition, although our elucidation of the crystal structure of the TTBK2 kinase domain indicates almost identical structural similarity with TTBK1, biochemical and cellular assays demonstrate that the enzymatic activity of these two proteins is regulated by a combination of unique extra-catalytic sequences and autophosphorylation events. Finally, we have identified an unbiased list of neuronal interactors and phosphorylation substrates for TTBK1 and TTBK2 that highlight the unique cellular pathways and functional networks that each isoform is involved in. This data address an important gap in knowledge regarding the implications of targeting TTBK kinases and may prove valuable in the development of potential therapies for disease.

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Data Availability

All data and material described in this manuscript are available upon request. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE [1] partner repository with the following dataset identifiers: “Identification of TTBK1 autophosphorylation sites in HEK293 cells” PXD017939. “TTBK1/TTBK2 Phosphoproteomics in mouse cortical neurons” PXD017938. “Identifying interactors through BioID2 in rat cortical neurons” PXD017937.

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Funding

Biogen provided support in the form of salaries for all authors, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘Author Contributions’ section.

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  1. Channa Bao and Bekim Bajrami are Co-first authors and also have contributed equally to the work.

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  1. Biogen, Cambridge, MA, 02134, USA

    Channa Bao, Bekim Bajrami, Douglas J. Marcotte, Jayanth V. Chodaparambil, Hannah M. Kerns, Jaclyn Henderson, Ru Wei, Benbo Gao & Gregory M. Dillon

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  1. Channa Bao
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  2. Bekim Bajrami
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All experiments were conducted in compliance with the rules set forth by the Biogen Institutional Animal Use and Care Committee under protocol number 881 and in accordance with the guidelines established in the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Bao, C., Bajrami, B., Marcotte, D.J. et al. Mechanisms of Regulation and Diverse Activities of Tau-Tubulin Kinase (TTBK) Isoforms. Cell Mol Neurobiol 41, 669–685 (2021). https://doi.org/10.1007/s10571-020-00875-6

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