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Plant Cell Advance Online Publication
Published on January 19, 2005; 10.1105/tpc.104.028522

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Received October 11, 2004
Accepted November 12, 2004

Phosphoserines on Maize CENTROMERIC HISTONE H3 and Histone H3 Demarcate the Centromere and Pericentromere during Chromosome Segregation

Xiaolan Zhang 1, Xuexian Li 1, Joshua B. Marshall 1, Cathy X. Zhong 1, and R. Kelly Dawe 2*

1 Department of Plant Biology, University of Georgia, Athens, Georgia 30602
2 Department of Plant Biology, University of Georgia, Athens, Georgia 30602; Department of Genetics, University of Georgia, Athens, Georgia 30602

* To whom correspondence should be addressed. E-mail: kelly{at}plantbio.uga.edu.

We have identified and characterized a 17- to 18-kD Ser50-phosphorylated form of maize (Zea mays) CENTROMERIC HISTONE H3 (phCENH3-Ser50). Immunostaining in both mitosis and meiosis indicates that CENH3-Ser50 phosphorylation begins in prophase/diplotene, increases to a maximum at prometaphase-metaphase, and drops during anaphase. Dephosphorylation is precipitous (approximately sixfold) at the metaphase-anaphase transition, suggesting a role in the spindle checkpoint. Although phCENH3-Ser50 lies within a region that lacks homology to any other known histone, its closest counterpart is the phospho-Ser28 residue of histone H3 (phH3-Ser28). CENH3-Ser50 and H3-Ser28 are phosphorylated with nearly identical kinetics, but the former is restricted to centromeres and the latter to pericentromeres. Opposing centromeres separate in prometaphase, whereas the phH3-Ser28-marked pericentromeres remain attached and coalesce into a well-defined tether that binds the centromeres together. We propose that a centromere-initiated wave of histone phosphorylation is an early step in defining the two major structural domains required for chromosome segregation: centromere (alignment, motility) and pericentromere (cohesion).






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