This study identifies tubacin, a small-molecule inhibitor of histone deacetylase 6 (HDAC6)-mediated α-tubulin deacetylation. Tubacin selectively inhibits α-tubulin deacetylation without affecting histone acetylation, gene expression, or cell-cycle progression. It targets the catalytic domain of HDAC6, which is responsible for tubulin deacetylation. Tubacin treatment decreases cell motility but does not stabilize microtubules. Overexpression of HDAC6 disrupts the localization of p53, a protein involved in microtubule-Golgi interactions. These findings highlight the role of α-tubulin acetylation in microtubule-associated protein localization and suggest that HDAC6 inhibitors like tubacin may have therapeutic applications as antimetastatic and antiangiogenic agents. The study also shows that HDAC6 is a key player in α-tubulin acetylation, which is important for cell motility and processes such as protein trafficking. Tubacin's ability to selectively inhibit HDAC6-mediated α-tubulin deacetylation provides a new tool for studying the effects of deacetylase inhibitors on the cytoskeleton and chromatin. The results suggest that increasing α-tubulin acetylation may be important for the antimetastatic and antiangiogenic properties of HDAC inhibitors, while decreasing α-tubulin acetylation may contribute to the reduced acetylation levels observed in neurodegenerative disorders. The study also demonstrates that HDAC inhibitors like tubacin can be used to uncouple the effects of deacetylase inhibitors on the cytoskeleton and chromatin, allowing for new applications of deacetylase inhibitors with increased selectivity.This study identifies tubacin, a small-molecule inhibitor of histone deacetylase 6 (HDAC6)-mediated α-tubulin deacetylation. Tubacin selectively inhibits α-tubulin deacetylation without affecting histone acetylation, gene expression, or cell-cycle progression. It targets the catalytic domain of HDAC6, which is responsible for tubulin deacetylation. Tubacin treatment decreases cell motility but does not stabilize microtubules. Overexpression of HDAC6 disrupts the localization of p53, a protein involved in microtubule-Golgi interactions. These findings highlight the role of α-tubulin acetylation in microtubule-associated protein localization and suggest that HDAC6 inhibitors like tubacin may have therapeutic applications as antimetastatic and antiangiogenic agents. The study also shows that HDAC6 is a key player in α-tubulin acetylation, which is important for cell motility and processes such as protein trafficking. Tubacin's ability to selectively inhibit HDAC6-mediated α-tubulin deacetylation provides a new tool for studying the effects of deacetylase inhibitors on the cytoskeleton and chromatin. The results suggest that increasing α-tubulin acetylation may be important for the antimetastatic and antiangiogenic properties of HDAC inhibitors, while decreasing α-tubulin acetylation may contribute to the reduced acetylation levels observed in neurodegenerative disorders. The study also demonstrates that HDAC inhibitors like tubacin can be used to uncouple the effects of deacetylase inhibitors on the cytoskeleton and chromatin, allowing for new applications of deacetylase inhibitors with increased selectivity.