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Deacetylation of α-tubulin and cortactin is required for HDAC6 to trigger ciliary disassembly.

Ran J, Yang Y, Li D, Liu M, Zhou J - Sci Rep (2015)

Bottom Line: Ciliary dysfunction is associated with a variety of diseases known as ciliopathies.Overexpression of HDAC6 decreases the levels of acetylated α-tubulin and cortactin without affecting the expression or localization of known ciliary regulators.These findings provide mechanistic insight into the ciliary role of HDAC6 and underscore the importance of reversible acetylation in regulating ciliary homeostasis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin 300071, China.

ABSTRACT
Cilia play important roles in sensing extracellular signals and directing fluid flow. Ciliary dysfunction is associated with a variety of diseases known as ciliopathies. Histone deacetylase 6 (HDAC6) has recently emerged as a major driver of ciliary disassembly, but little is known about the downstream players. Here we provide the first evidence that HDAC6-mediated deacetylation of α-tubulin and cortactin is critical for its induction of ciliary disassembly. HDAC6 is localized in the cytoplasm and enriched at the centrosome and basal body. Overexpression of HDAC6 decreases the levels of acetylated α-tubulin and cortactin without affecting the expression or localization of known ciliary regulators. We also find that overexpression of α-tubulin or cortactin or their acetylation-deficient mutants enhances the ability of HDAC6 to induce ciliary disassembly. In addition, acetylation-mimicking mutants of α-tubulin and cortactin counteract HDAC6-induced ciliary disassembly. Furthermore, HDAC6 stimulates actin polymerization, and inhibition of actin polymerization abolishes the activity of HDAC6 to trigger ciliary disassembly. These findings provide mechanistic insight into the ciliary role of HDAC6 and underscore the importance of reversible acetylation in regulating ciliary homeostasis.

No MeSH data available.


HDAC6 stimulates actin polymerization.(A) Immunofluorescence images of RPE-1 cells transfected with GFP vector or GFP-HDAC6, untreated or treated with tubacin (2 μM) or vehicle control, serum-starved for 24 hours, and stained with phalloidin and DAPI. Scale bar, 5 μm. (B) Experiments were performed as in A, and F-actin intensity was quantified. **P < 0.01, ***P < 0.001; ns, not significant. Error bars indicate SEM.
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f8: HDAC6 stimulates actin polymerization.(A) Immunofluorescence images of RPE-1 cells transfected with GFP vector or GFP-HDAC6, untreated or treated with tubacin (2 μM) or vehicle control, serum-starved for 24 hours, and stained with phalloidin and DAPI. Scale bar, 5 μm. (B) Experiments were performed as in A, and F-actin intensity was quantified. **P < 0.01, ***P < 0.001; ns, not significant. Error bars indicate SEM.

Mentions: Since HDAC6 deacetylates cortactin and thereby increases the interaction of cortactin with F-actin20, we speculated that HDAC6 might promote actin polymerization. To test this possibility, RPE1 cells were transfected with GFP-HDAC6, and F-actin was stained with phalloidin. We found that transfection of GFP-HDAC6 significantly increased F-actin intensity, as compared with transfection with GFP vector (Fig. 8A,B). In addition, the HDAC6 inhibitor tubacin suppressed HDAC6-induced increase of F-actin staining (Fig. 8A,B). These data indicate that HDAC6 overexpression stimulates actin polymerization.


Deacetylation of α-tubulin and cortactin is required for HDAC6 to trigger ciliary disassembly.

Ran J, Yang Y, Li D, Liu M, Zhou J - Sci Rep (2015)

HDAC6 stimulates actin polymerization.(A) Immunofluorescence images of RPE-1 cells transfected with GFP vector or GFP-HDAC6, untreated or treated with tubacin (2 μM) or vehicle control, serum-starved for 24 hours, and stained with phalloidin and DAPI. Scale bar, 5 μm. (B) Experiments were performed as in A, and F-actin intensity was quantified. **P < 0.01, ***P < 0.001; ns, not significant. Error bars indicate SEM.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526867&req=5

f8: HDAC6 stimulates actin polymerization.(A) Immunofluorescence images of RPE-1 cells transfected with GFP vector or GFP-HDAC6, untreated or treated with tubacin (2 μM) or vehicle control, serum-starved for 24 hours, and stained with phalloidin and DAPI. Scale bar, 5 μm. (B) Experiments were performed as in A, and F-actin intensity was quantified. **P < 0.01, ***P < 0.001; ns, not significant. Error bars indicate SEM.
Mentions: Since HDAC6 deacetylates cortactin and thereby increases the interaction of cortactin with F-actin20, we speculated that HDAC6 might promote actin polymerization. To test this possibility, RPE1 cells were transfected with GFP-HDAC6, and F-actin was stained with phalloidin. We found that transfection of GFP-HDAC6 significantly increased F-actin intensity, as compared with transfection with GFP vector (Fig. 8A,B). In addition, the HDAC6 inhibitor tubacin suppressed HDAC6-induced increase of F-actin staining (Fig. 8A,B). These data indicate that HDAC6 overexpression stimulates actin polymerization.

Bottom Line: Ciliary dysfunction is associated with a variety of diseases known as ciliopathies.Overexpression of HDAC6 decreases the levels of acetylated α-tubulin and cortactin without affecting the expression or localization of known ciliary regulators.These findings provide mechanistic insight into the ciliary role of HDAC6 and underscore the importance of reversible acetylation in regulating ciliary homeostasis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin 300071, China.

ABSTRACT
Cilia play important roles in sensing extracellular signals and directing fluid flow. Ciliary dysfunction is associated with a variety of diseases known as ciliopathies. Histone deacetylase 6 (HDAC6) has recently emerged as a major driver of ciliary disassembly, but little is known about the downstream players. Here we provide the first evidence that HDAC6-mediated deacetylation of α-tubulin and cortactin is critical for its induction of ciliary disassembly. HDAC6 is localized in the cytoplasm and enriched at the centrosome and basal body. Overexpression of HDAC6 decreases the levels of acetylated α-tubulin and cortactin without affecting the expression or localization of known ciliary regulators. We also find that overexpression of α-tubulin or cortactin or their acetylation-deficient mutants enhances the ability of HDAC6 to induce ciliary disassembly. In addition, acetylation-mimicking mutants of α-tubulin and cortactin counteract HDAC6-induced ciliary disassembly. Furthermore, HDAC6 stimulates actin polymerization, and inhibition of actin polymerization abolishes the activity of HDAC6 to trigger ciliary disassembly. These findings provide mechanistic insight into the ciliary role of HDAC6 and underscore the importance of reversible acetylation in regulating ciliary homeostasis.

No MeSH data available.