Phosphorylation-induced rearrangement of the histone H3 NH2-terminal domain during mitotic chromosome condensation.
The NH2-terminal domain (N-tail) of histone H3 has been implicated in chromatin compaction and its phosphorylation at Ser10 is tightly correlated with mitotic chromosome condensation. We have developed one mAb that specifically recognizes histone H3 N-tails phosphorylated at Ser10 (H3P Ab) and another that recognizes phosphorylated and unphosphorylated H3 N-tails equally well (H3 Ab). Immunocytochemistry with the H3P Ab shows that Ser10 phosphorylation begins in early prophase, peaks before metaphase, and decreases during anaphase and telophase. Unexpectedly, the H3 Ab shows stronger immunofluorescence in mitosis than interphase, indicating that the H3 N-tail is more accessible in condensed mitotic chromatin than in decondensed interphase chromatin. In vivo ultraviolet laser cross-linking indicates that the H3 N-tail is bound to DNA in interphase cells and that binding is reduced in mitotic cells. Treatment of mitotic cells with the protein kinase inhibitor staurosporine causes histone H3 dephosphorylation and chromosome decondensation. It also decreases the accessibility of the H3 N-tail to H3 Ab and increases the binding of the N-tail to DNA. These results indicate that a phosphorylation-dependent weakening of the association between the H3 N-tail and DNA plays a role in mitotic chromosome condensation.
Sauvé
D M
DM
Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
Anderson
H J
HJ
Ray
J M
JM
James
W M
WM
Roberge
M
M
D000705:Anaphase; D000911:Antibodies, Monoclonal; D001943:Breast Neoplasms; D002453:Cell Cycle / Q000502:physiology*; D002877:Chromosomes, Human / Q000502:physiology* / Q000648:ultrastructure*; D005260:Female; D006657:Histones / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D007399:Interphase; D008677:Metaphase; D008938:Mitosis; D010766:Phosphorylation; D011418:Prophase; D006656:Protamine Kinase / Q000378:metabolism; D012694:Serine; D013096:Spermine / Q000378:metabolism; D019311:Staurosporine / Q000494:pharmacology; D013692:Telophase; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D006657:Histones; D013096:Spermine; D012694:Serine; D006656:Protamine Kinase; D019311:Staurosporine