Histone deacetylase inhibitors all induce p21 but differentially cause tubulin acetylation, mitotic arrest, and cytotoxicity.
By preventing deacetylation of histones, histone deacetylase inhibitors (HDIs) transcriptionally induce p21. Here we show that the HDIs sodium butyrate (Bu), trichostatin A (TSA) and depsipeptide (FR901228) all induced p21, but only TSA and FR901228 caused mitotic arrest (in addition to arrest in G1 and G2). The ability to cause mitotic arrest correlated with the higher cytotoxicity of these compounds. Although causing mitotic arrest, TSA and FR901228 (unlike paclitaxel) did not affect tubulin polymerization. Unlike FR9012208, TSA caused acetylation of tubulin at lysine 40; both soluble tubulin and microtubules were acetylated. Whereas the induction of p21 reached a maximum by 8 h, tubulin was maximally acetylated after only 1 h of TSA treatment. Tubulin acetylation was detectable after treatment with 12-25 ng/ml TSA although acetylation plateaued at 50 ng/ml TSA, coinciding with G2-M arrest, appearance of cells with a sub-2N DNA content, poly(ADP-ribose) polymerase cleavage, and rapid cell death. We conclude that HDIs have differential effects on non-histone deacetylases and that rapid acetylation of tubulin caused by TSA is a marker of nontranscriptional effects of TSA.
Blagosklonny
Mikhail V
MV
Brander Cancer Research Institute, New York Medical College, Valhalla, New York 10595, USA. M_Blagosklonny@NYMC.EDU
Robey
Robert
R
Sackett
Dan L
DL
Du
Litong
L
Traganos
Frank
F
Darzynkiewicz
Zbigniew
Z
Fojo
Tito
T
Bates
Susan E
SE
D018686:Anesthetics, Intravenous / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis; D002087:Butyrates / Q000494:pharmacology; D002453:Cell Cycle; D002460:Cell Line; D004396:Coloring Agents / Q000494:pharmacology; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D047630:Depsipeptides; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D016195:G2 Phase; D018922:HL-60 Cells; D056572:Histone Deacetylase Inhibitors; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D015151:Immunoblotting; D019169:Jurkat Cells; D008938:Mitosis; D010455:Peptides / Q000494:pharmacology; D010456:Peptides, Cyclic; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012978:Sodium Oxybate / Q000494:pharmacology; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D013997:Time Factors; D014158:Transcription, Genetic; D014404:Tubulin / Q000378:metabolism; D014407:Tumor Cells, Cultured
D018686:Anesthetics, Intravenous; D000900:Anti-Bacterial Agents; D000903:Antibiotics, Antineoplastic; D002087:Butyrates; C493956:CDKN1A protein, human; D004396:Coloring Agents; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D047630:Depsipeptides; D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D010455:Peptides; D010456:Peptides, Cyclic; D011500:Protein Synthesis Inhibitors; D013778:Tetrazolium Salts; D013844:Thiazoles; D014404:Tubulin; C012589:trichostatin A; D012978:Sodium Oxybate; D004247:DNA; C087123:romidepsin; C022616:thiazolyl blue