Inhibitors of histone deacetylases alter kinetochore assembly by disrupting pericentromeric heterochromatin.
The kinetochore, a multi-protein complex assembled on centromeric chromatin in mitosis, is essential for sister chromosome segregation. We show here that inhibition of histone deacetylation blocks mitotic progression at prometaphase in two human tumor cell lines by interfering with kinetochore assembly. Decreased amounts of hBUB1, CENP-F and the motor protein CENP-E were present on kinetochores of treated cells. These kinetochores failed to nucleate and inefficiently captured microtubules, resulting in activation of the mitotic checkpoint. Addition of histone deacetylase inhibitors prior to the end of S-phase resulted in decreased HP1-beta on pericentromeric heterochromatin in S-phase and G(2), decreased pericentromeric targeting of Aurora B kinase, resulting in decreased premitotic phosphorylation of pericentromeric histone H3(S10) in G(2), followed by assembly of deficient kinetochores in M-phase. HP1-beta, Aurora B and the affected kinetochore proteins all were present at normal levels in treated cells; thus, effects of the inhibitors on mitotic progression do not seem to reflect changes in gene expression. In vitro kinase activity of Aurora B isolated from treated cells was unaffected. We propose that the increased presence in pericentromeric heterochromatin of histone H3 acetylated at K9 is responsible for the mitotic defects resulting from inhibition of histone deacetylation.
Robbins
April R
AR
Laboratory of Cell Biochemistry and Biology, National Institute of Diabetes, Digestive and Kidney Diseases, Bethesda, Maryland 20892, USA. cwharr@helix.nih.gov
Jablonski
Sandra A
SA
Yen
Tim J
TJ
Yoda
Kinya
K
Robey
Rob
R
Bates
Susan E
SE
Sackett
Dan L
DL
D000107:Acetylation; D064107:Aurora Kinase B; D064127:Aurora Kinases; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D002503:Centromere / Q000187:drug effects / Q000502:physiology*; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis / Q000378:metabolism; D020090:Chromosome Segregation / Q000187:drug effects; D047630:Depsipeptides / Q000494:pharmacology; D016195:G2 Phase / Q000187:drug effects / Q000502:physiology; D006570:Heterochromatin / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D018386:Kinetochores / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D008387:Marine Toxins; D008727:Methotrexate / Q000494:pharmacology; D052998:Microcystins; D008840:Microfilament Proteins; D008938:Mitosis / Q000187:drug effects / Q000502:physiology; D010456:Peptides, Cyclic / Q000494:pharmacology; D011494:Protein Kinases / Q000032:analysis; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000032:analysis / Q000187:drug effects / Q000502:physiology; D016196:S Phase / Q000187:drug effects / Q000502:physiology; D008941:Spindle Apparatus / Q000187:drug effects / Q000502:physiology
D002868:Chromosomal Proteins, Non-Histone; D047630:Depsipeptides; D006570:Heterochromatin; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; D007211:Indoles; D008387:Marine Toxins; D052998:Microcystins; D008840:Microfilament Proteins; D010456:Peptides, Cyclic; C068688:centromere protein E; C094772:centromere protein F; D000090266:Chromobox Protein Homolog 5; C012589:trichostatin A; C007293:DAPI; C037654:HC toxin; C087123:romidepsin; D011494:Protein Kinases; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; C584277:BUB1 protein, human; C090464:Bub1 spindle checkpoint protein; D017346:Protein Serine-Threonine Kinases; D006655:Histone Deacetylases; C057862:cyanoginosin LR; D008727:Methotrexate