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

J 2020

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

USTINOVA, Kseniya; Zora NOVAKOVA; Makoto SAITO; Marat MELESHIN; Jana MIKESOVA et al.

Základní údaje

Originální název

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

Autoři

USTINOVA, Kseniya; Zora NOVAKOVA; Makoto SAITO; Marat MELESHIN; Jana MIKESOVA; Zsofia KUTIL; Petra BARANOVA; Barbora HAVLINOVA; Mike SCHUTKOWSKI; Patrick MATTHIAS a Cyril BARINKA

Vydání

Journal of Biological Chemistry, Bethesda, Amer. Soc. Biochem. Mol. Biol. 2020, 0021-9258

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 5.157

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:90043/20:00139203

Organizační jednotka

CIISB

Klíčová slova anglicky

structure-function; substrate specificity; protein-protein interaction; microtubule-associated protein (MAP); tubulin; intrinsically disordered protein; histone deacetylase 6 (HDAC6); protein motif

Štítky

ne MU, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 28. 3. 2025 16:39, Mgr. Eva Dubská

Anotace

V originále

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.

Návaznosti

90043, velká výzkumná infrastruktura

Název: CIISB

Citovat

USTINOVA, Kseniya; Zora NOVAKOVA; Makoto SAITO; Marat MELESHIN; Jana MIKESOVA; Zsofia KUTIL; Petra BARANOVA; Barbora HAVLINOVA; Mike SCHUTKOWSKI; Patrick MATTHIAS a Cyril BARINKA. The disordered N-terminus of HDAC6 is a microtubule-binding domain critical for efficient tubulin deacetylation. Journal of Biological Chemistry. Bethesda: Amer. Soc. Biochem. Mol. Biol., 2020, roč. 295, č. 9, s. 2614-2628. ISSN 0021-9258. Dostupné z: https://doi.org/10.1074/jbc.RA119.011243.

@article{2487288,
   author = {Ustinova, Kseniya and Novakova, Zora and Saito, Makoto and Meleshin, Marat and Mikesova, Jana and Kutil, Zsofia and Baranova, Petra and Havlinova, Barbora and Schutkowski, Mike and Matthias, Patrick and Barinka, Cyril},
   article_location = {Bethesda},
   article_number = {9},
   doi = {https://doi.org/10.1074/jbc.RA119.011243},
   keywords = {structure-function; substrate specificity; protein-protein interaction; microtubule-associated protein (MAP); tubulin; intrinsically disordered protein; histone deacetylase 6 (HDAC6); protein motif},
   language = {eng},
   issn = {0021-9258},
   journal = {Journal of Biological Chemistry},
   title = {The disordered N-terminus of HDAC6 is a microtubule-binding domain critical for efficient tubulin deacetylation},
   url = {https://www.sciencedirect.com/science/article/pii/S0021925817490004?via%3Dihub},
   volume = {295},
   year = {2020}
}

TY  - JOUR
ID  - 2487288
AU  - Ustinova, Kseniya - Novakova, Zora - Saito, Makoto - Meleshin, Marat - Mikesova, Jana - Kutil, Zsofia - Baranova, Petra - Havlinova, Barbora - Schutkowski, Mike - Matthias, Patrick - Barinka, Cyril
PY  - 2020
TI  - The disordered N-terminus of HDAC6 is a microtubule-binding domain critical for efficient tubulin deacetylation
JF  - Journal of Biological Chemistry
VL  - 295
IS  - 9
SP  - 2614-2628
EP  - 2614-2628
PB  - Amer. Soc. Biochem. Mol. Biol.
SN  - 00219258
KW  - structure-function
KW  - substrate specificity
KW  - protein-protein interaction
KW  - microtubule-associated protein (MAP)
KW  - tubulin
KW  - intrinsically disordered protein
KW  - histone deacetylase 6 (HDAC6)
KW  - protein motif
UR  - https://www.sciencedirect.com/science/article/pii/S0021925817490004?via%3Dihub
N2  - 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.
ER  -

USTINOVA, Kseniya; Zora NOVAKOVA; Makoto SAITO; Marat MELESHIN; Jana MIKESOVA; Zsofia KUTIL; Petra BARANOVA; Barbora HAVLINOVA; Mike SCHUTKOWSKI; Patrick MATTHIAS a Cyril BARINKA. The disordered N-terminus of HDAC6 is a microtubule-binding domain critical for efficient tubulin deacetylation. \textit{Journal of Biological Chemistry}. Bethesda: Amer. Soc. Biochem. Mol. Biol., 2020, roč.~295, č.~9, s.~2614-2628. ISSN~0021-9258. Dostupné z: https://doi.org/10.1074/jbc.RA119.011243.

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