Ataxin-3 (AT3) is the protein that triggers the inherited neurodegenerative disorder spinocerebellar ataxia type 3 when its polyglutamine (polyQ) stretch close to the C-terminus exceeds a critical length. AT3 consists of the N-terminal globular Josephin domain (JD) and the C-terminal disordered one. It cleaves isopeptide bonds between ubiquitin monomers, an event involved in protein quality control mechanisms. AT3 has been implicated in the pathway that sorts aggregated protein to aggresomes via microtubules, in which dynein and histone deacetylase 6 (HDAC6) also seem to be involved. By taking advantage of small angle X-ray scattering (SAXS) and surface plasmon resonance (SPR), we have investigated the interaction of AT3 with tubulin and HDAC6. Based on SAXS results, the AT3 oligomer, consisting of 6-7 subunits, tightly binds to the tubulin hexameric oligomer in a "parallel" fashion. By SPR analysis we have demonstrated that AT3 binds to tubulin dimer with a 50nM affinity. Binding fits with a Langmuir 1:1 model and involves a single binding interface. Nevertheless, the interaction surface consists of three distinct, discontinuous tubulin-binding regions (TBR), one located in the JD, and the two others in the disordered domain, upstream and downstream of the polyQ stretch. In the absence of any of the three TBRs, the affinity is drastically reduced. By SPR we have also provided the first evidence of direct binding of AT3 to HDAC6, with affinity in the range 0.1-1μM. These results shed light on the interactions among the components of the transport machinery that sorts aggregate protein to the aggresome, and pave the way to in vivo studies aimed at further clarifying their roles.

Bonanomi, M., Mazzucchelli, S., D'Urzo, A., Nardini, M., Konarev, P., Invernizzi, G., et al. (2014). Interactions of ataxin-3 with its molecular partners in the protein machinery that sorts protein aggregates to the aggresome. THE INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY, 51(1), 58-64 [10.1016/j.biocel.2014.03.015].

Interactions of ataxin-3 with its molecular partners in the protein machinery that sorts protein aggregates to the aggresome

BONANOMI, MARCELLA;MAZZUCCHELLI, SERENA;D'URZO, ANNALISA;INVERNIZZI, GAETANO;VANONI, MARCO ERCOLE;REGONESI, MARIA ELENA;TORTORA, PAOLO
2014

Abstract

Ataxin-3 (AT3) is the protein that triggers the inherited neurodegenerative disorder spinocerebellar ataxia type 3 when its polyglutamine (polyQ) stretch close to the C-terminus exceeds a critical length. AT3 consists of the N-terminal globular Josephin domain (JD) and the C-terminal disordered one. It cleaves isopeptide bonds between ubiquitin monomers, an event involved in protein quality control mechanisms. AT3 has been implicated in the pathway that sorts aggregated protein to aggresomes via microtubules, in which dynein and histone deacetylase 6 (HDAC6) also seem to be involved. By taking advantage of small angle X-ray scattering (SAXS) and surface plasmon resonance (SPR), we have investigated the interaction of AT3 with tubulin and HDAC6. Based on SAXS results, the AT3 oligomer, consisting of 6-7 subunits, tightly binds to the tubulin hexameric oligomer in a "parallel" fashion. By SPR analysis we have demonstrated that AT3 binds to tubulin dimer with a 50nM affinity. Binding fits with a Langmuir 1:1 model and involves a single binding interface. Nevertheless, the interaction surface consists of three distinct, discontinuous tubulin-binding regions (TBR), one located in the JD, and the two others in the disordered domain, upstream and downstream of the polyQ stretch. In the absence of any of the three TBRs, the affinity is drastically reduced. By SPR we have also provided the first evidence of direct binding of AT3 to HDAC6, with affinity in the range 0.1-1μM. These results shed light on the interactions among the components of the transport machinery that sorts aggregate protein to the aggresome, and pave the way to in vivo studies aimed at further clarifying their roles.
Articolo in rivista - Articolo scientifico
Small angle X-ray scattering; Ataxin-3; Surface plasmon resonance; Tubulin dimer; HDAC6
English
2014
51
1
58
64
none
Bonanomi, M., Mazzucchelli, S., D'Urzo, A., Nardini, M., Konarev, P., Invernizzi, G., et al. (2014). Interactions of ataxin-3 with its molecular partners in the protein machinery that sorts protein aggregates to the aggresome. THE INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY, 51(1), 58-64 [10.1016/j.biocel.2014.03.015].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/51958
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