The disordered N-terminus of HDAC6 is a microtubule-binding domain critical for efficient tubulin deacetylation

0524504 - BTÚ 2021 RIV US eng J - Journal Article
Ustinova, Kseniya - Nováková, Zora - Saito, M. - Meleshin, M. - Mikesova, Jana - Kutil, Zsofia - Baranová, Petra - Havlínová, Barbora - Schutkowski, M. - Matthias, P. - Bařinka, Cyril
The disordered N-terminus of HDAC6 is a microtubule-binding domain critical for efficient tubulin deacetylation.
Journal of Biological Chemistry. Roč. 295, č. 9 (2020), s. 2614-2628. ISSN 0021-9258. E-ISSN 1083-351X
R&D Projects: GA MŠMT(CZ) LM2015062; GA ČR GA15-19640S; GA MŠMT(CZ) LM2015043; GA MŠMT(CZ) ED1.1.00/02.0109
Institutional support: RVO:86652036
Keywords : structure-function * protein-protein interaction * substrate specificity
OECD category: Biochemistry and molecular biology
Impact factor: 5.157, year: 2020
Method of publishing: Open access
https://www.jbc.org/content/295/9/2614.full#ack-1

Histone deacetylase 6 (HDAC6) is a multidomain cytosolic enzyme having tubulin deacetylase activity that has been unequivocally assigned to the second of the tandem catalytic domains. However, virtually no information exists on the contribution of other HDAC6 domains on tubulin recognition. Here, using recombinant protein expression, site-directed mutagenesis, fluorimetric and biochemical assays, microscale thermophoresis, and total internal reflection fluorescence microscopy, we identified the N-terminal, disordered region of HDAC6 as a microtubule-binding domain and functionally characterized it to the single-molecule level. We show that the microtubule-binding motif spans two positively charged patches comprising residues Lys-32 to Lys-58. We found that HDAC6-microtubule interactions are entirely independent of the catalytic domains and are mediated by ionic interactions with the negatively charged microtubule surface. Importantly, a crosstalk between the microtubule-binding domain and the deacetylase domain was critical for recognition and efficient deacetylation of free tubulin dimers both in vitro and in vivo. Overall, our results reveal that recognition of substrates by HDAC6 is more complex than previously appreciated and that domains outside the tandem catalytic core are essential for proficient substrate deacetylation.
Permanent Link: http://hdl.handle.net/11104/0308861