Visualizing histone modifications in living cells: spatiotemporal dynamics of H3 phosphorylation during interphase.
Posttranslational histone modifications regulate both gene expression and genome integrity. Despite the dynamic nature of these modifications, appropriate real-time monitoring systems are lacking. In this study, we developed a method to visualize histone modifications in living somatic cells and preimplantation embryos by loading fluorescently labeled specific Fab antibody fragments. The technique was used to study histone H3 Ser10 (H3S10) phosphorylation, which occurs during chromosome condensation in mitosis mediated by the aurora B kinase. In aneuploid cancer cells that frequently missegregate chromosomes, H3S10 is phosphorylated just before the chromosomes condense, whereas aurora B already accumulates in nuclei during S phase. In contrast, in nontransformed cells, phosphorylated H3S10 foci appear for a few hours during interphase, and transient exposure to an aurora B-selective inhibitor during this period induces chromosome missegregation. These results suggest that, during interphase, moderate aurora B activity or H3S10 phosphorylation is required for accurate chromosome segregation. Visualizing histone modifications in living cells will facilitate future epigenetic and cell regulation studies.
Hayashi-Takanaka
Yoko
Y
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
Yamagata
Kazuo
K
Nozaki
Naohito
N
Kimura
Hiroshi
H
D000818:Animals; D064107:Aurora Kinase B; D064127:Aurora Kinases; D001755:Blastocyst / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects; D020090:Chromosome Segregation; D005347:Fibroblasts / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching; D006367:HeLa Cells; D006657:Histones / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D007399:Interphase / Q000187:drug effects; D051379:Mice; D018613:Microscopy, Confocal; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D054645:Protein Phosphatase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D012694:Serine; D013997:Time Factors
D006657:Histones; D007140:Immunoglobulin Fab Fragments; D047428:Protein Kinase Inhibitors; D012694:Serine; C576921:AURKB protein, human; C576924:Aurkb protein, mouse; D064107:Aurora Kinase B; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; D054645:Protein Phosphatase 1