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

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

  • Ustinova K Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50 Vestec, Czech Republic.
  • Novakova Z Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50 Vestec, Czech Republic.
  • Saito M Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland.
  • Meleshin M Department of Enzymology, Institute of Biochemistry and Biotechnology, Charles Tanford Protein Center, Martin Luther University, Halle-Wittenberg, 06120 Halle/Saale, Germany.
  • Mikesova J Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50 Vestec, Czech Republic.
  • Kutil Z Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50 Vestec, Czech Republic.
  • Baranova P Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50 Vestec, Czech Republic.
  • Havlinova B Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50 Vestec, Czech Republic.
  • Schutkowski M Department of Enzymology, Institute of Biochemistry and Biotechnology, Charles Tanford Protein Center, Martin Luther University, Halle-Wittenberg, 06120 Halle/Saale, Germany.
  • Matthias P Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland.
  • Barinka C Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 252 50 Vestec, Czech Republic cyril.barinka@ibt.cas.cz.
Show more…
  • 2020-01-19
Published in:
  • The Journal of biological chemistry. - 2020
cytoskeleton
histone deacetylase 6 (HDAC6)
intrinsically disordered protein
microtubule-associated protein (MAP)
post-translational modification
protein motif
protein-protein interaction
structure-function
substrate specificity
total internal reflection fluorescence (TIRF)
tubulin
Acetylation
Amino Acid Sequence
Catalytic Domain
Histone Deacetylase 6
Humans
Microtubules
Protein Binding
Protein Domains
Substrate Specificity
Tubulin
English 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.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/139150
Statistics
Document views: 10 File downloads: