Biochemical and Biophysical Research Communications
Effects of downregulated HDAC6 expression on the proliferation of lung cancer cells
Section snippets
Materials and methods
Reagents. Mouse anti-α-tubulin (B-5-1-2) and anti-acetylated (Ac-) α-tubulin (6-11B-1) antibodies were purchased from Sigma–Aldrich. Rabbit anti-detyrosinated (Glu-) α-tubulin antibodies were a generous gift from Dr. T.H. MacRae (Department of Biology, Dalhousie University, Canada). Mouse anti-ubiquitin (P4D1), anti-EGFR (528, for immunoprecipitation), rabbit anti-EGFR (1005, for Western blotting), anti-PDGFRα, (951), anti-HDAC6 (H-300), anti-cyclin A (H-432), anti-CDK2 (M2), and anti-CDK4
HDAC6 regulates the polymerization of microtubules
Using a pharmacologic approach, we previously demonstrated that HDAC6 promotes instability of a dynamic pool of MTs [2]. To confirm this using an alternative approach, we first examined the polymerized/depolymerized ratio of MTs in cells in which HDAC6 expression was stably knocked down (HDAC6-KD cells). As shown in Fig. 1A, the level of Ac-α-tubulin was significantly increased in HDAC6-KD cells compared to control cells (pS cells), whereas cells stably expressing wild-type HDAC6 (WT OP cells)
Acknowledgments
This work was supported in part by the CREST Research Project, the Japan Science and Technology Corporation, and a Grant-in-Aid for Scientific Research on Priority Area “Cancer” from the Ministry of Education, Culture, Sports, Science and Technology of Japan. The laboratory of S.K. is supported by CLARA cancéropôle (EpiPro, EpiMed).
We thank Dr. T.H. MacRae (Dalhousie University) for supplying the rabbit anti-Glu-α-tubulin antibodies and Dr. S. Rousseaux (S.K. Laboratory) for critically reading
References (26)
- et al.
Microtubule acetylation promotes kinesin-1 binding and transport
Curr. Biol.
(2006) - et al.
HDAC6 regulates Hsp90 acetylation and chaperone-dependent activation of glucocorticoid receptor
Mol. Cell
(2005) - et al.
Inhibition of histone deacetylase 6 acetylates and disrupts the chaperone function of heat shock protein 90: a novel basis of antileukemia activity of histone deacetylase inhibitors
J. Biol. Chem.
(2005) - et al.
The platelet-derived growth factor receptor α is destabilized by geldanamycins in cancer cells
J. Biol. Chem.
(2007) - et al.
Class II histone deacetylases play pivotal roles in heat shock protein 90-mediated proteasomal degradation of vascular endothelial growth factor receptors
Biochem. Biophys. Res. Commun.
(2008) - et al.
The deacetylase HDAC6 regulates aggresome formation and cell viability in response to misfolded protein stress
Cell
(2003) - et al.
Gleevec (STI-571) inhibits lung cancer cell growth (A549) and potentiates the cisplatin effect in vitro
Mol. Cancer
(2003) - et al.
Sensitivity of mature ErbB2 to geldanamycin is conferred by its kinase domain and is mediated by the chaperone protein Hsp90
J. Biol. Chem.
(2001) - et al.
HDAC6 is a microtubule-associated deacetylase
Nature
(2002) - et al.
In vivo destabilization of dynamic microtubules by HDAC6-mediated deacetylation
EMBO J.
(2002)
HDAC-6 interacts with and deacetylates tubulin and microtubules in vivo
EMBO J.
Posttranslational modification and microtubule stability
J. Cell Biol.
Microtubules containing acetylated α-tubulin in mammalian cells in culture
J. Cell Biol.
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Present address: Department of Bio-Science, Nagahama Institute of Bio-Science and Technology, Nagahama, Shiga 526-0829, Japan.