Two Catalytic Domains Are Required for Protein Deacetylation

Yu Zhang,Benoit Gilquin,Saadi Khochbin,Patrick Matthias

doi:10.1074/jbc.c500241200

Abstract

Histone deacetylase (HDAC)-6 was recently identified as a dual substrate, possibly multisubstrate, deacetylase that can act both on acetylated histone tails and on alpha-tubulin acetylated on Lys40. HDAC-6 is unique among deacetylases in having two hdac domains, and we have used this enzyme as a useful model to dissect the structural requirements for the deacetylation reaction. In this report, we show that both hdac domains are required for the intact deacetylase activity of HDAC-6 in vitro and in vivo. The spatial arrangement of these two domains in HDAC-6 is essential and alteration of the linker region between the two domains severely affects the catalytic activity. Artificial chimeric HDACs, made by replacing the hdac domains in HDAC-6 with corresponding domains from other class II HDACs, show de novo deacetylase activity. Taken together, our results demonstrate for the first time that the spatial arrangement of hdac domains is critical for in vivo deacetylation reaction and may provide a useful model for the development of novel HDAC inhibitors.

Highlights

Histone deacetylase (HDAC)-6 was recently identified as a dual substrate, possibly multisubstrate, deacetylase that can act both on acetylated histone tails and on ␣-tubulin acetylated on Lys40
We have shown previously that each hdac domain in HDAC-6 is sufficient to bind on its own ␤-tubulin, and this interaction is maintained even when the catalytic core is mutated [5]
Mutation analysis revealed that the ER motif regions of Esa1 or Rpd3 are required for histone acetyltransferases (HATs) activity of Esa1 and HDAC activity of Rpd3, respectively [7]

Summary

Introduction

Histone deacetylase (HDAC)-6 was recently identified as a dual substrate, possibly multisubstrate, deacetylase that can act both on acetylated histone tails and on ␣-tubulin acetylated on Lys. HDAC-6 is unique among deacetylases in having two hdac domains, and we have used this enzyme as a useful model to dissect the structural requirements for the deacetylation reaction. We show that both hdac domains are required for the intact deacetylase activity of HDAC-6 in vitro and in vivo. Among the HDACs, HDAC-6 was recently identified as a dual substrate, and possibly multisubstrate, deacetylase that can deacetylate both histone tails and ␣-tubulin Lys in vitro and in vivo [3, 4, 5]. In this report, using various in vitro and in vivo assays we demonstrate that both hdac domains of HDAC-6 are essential for activity of this enzyme and propose that the presence of more than one hdac domain is a general requirement for the deacetylation reaction

Methods

Results

Conclusion