Mitosis-specific phosphorylation of histone H3 initiates primarily within pericentromeric heterochromatin during G2 and spreads in an ordered fashion coincident with mitotic chromosome condensation.
We have generated and characterized a novel site-specific antibody highly specific for the phosphorylated form of the amino-terminus of histone H3 (Ser10). In this study, we used this antibody to examine in detail the relationship between H3 phosphorylation and mitotic chromosome condensation in mammalian cells. Our results extend previous biochemical studies by demonstrating that mitotic phosphorylation of H3 initiates nonrandomly in pericentromeric heterochromatin in late G2 interphase cells. Following initiation, H3 phosphorylation appears to spread throughout the condensing chromatin and is complete in most cell lines just prior to the formation of prophase chromosomes, in which a phosphorylated, but nonmitotic, chromosomal organization is observed. In general, there is a precise spatial and temporal correlation between H3 phosphorylation and initial stages of chromatin condensation. Dephosphorylation of H3 begins in anaphase and is complete immediately prior to detectable chromosome decondensation in telophase cells. We propose that the singular phosphorylation of the amino-terminus of histone H3 may be involved in facilitating two key functions during mitosis: (1) regulate protein-protein interactions to promote binding of trans-acting factors that "drive" chromatin condensation as cells enter M-phase and (2) coordinate chromatin decondensation associated with M-phase.
Hendzel
M J
MJ
Department of Anatomy and Medical Biochemistry, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada T2N 4N1.
Wei
Y
Y
Mancini
M A
MA
Van Hooser
A
A
Ranalli
T
T
Brinkley
B R
BR
Bazett-Jones
D P
DP
Allis
C D
CD
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D002460:Cell Line; D002503:Centromere / Q000737:chemistry; D016195:G2 Phase / Q000502:physiology*; D006570:Heterochromatin / Q000737:chemistry*; D006657:Histones / Q000032:analysis* / Q000378:metabolism; D007150:Immunohistochemistry; D008938:Mitosis / Q000502:physiology*; D008969:Molecular Sequence Data; D010748:Phosphopeptides / Q000032:analysis*; D010766:Phosphorylation; D011817:Rabbits
D006570:Heterochromatin; D006657:Histones; D010748:Phosphopeptides