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© The Rockefeller University Press, 0021-9525/1999//425 $5.00
The Journal of Cell Biology, Volume 145, Number 3, , 1999 425-435

The Maize Homologue of the Cell Cycle Checkpoint Protein MAD2 Reveals Kinetochore Substructure and Contrasting Mitotic and Meiotic Localization Patterns

Hong-Guo Yu^*, Michael G. Muszynski, and R. Kelly Dawe^*,

^* Department of Botany and Department of Genetics, University of Georgia, Athens, Georgia 30602; Pioneer Hi-Bred International, Inc., Johnston, Iowa 50131

We have identified a maize homologue of yeast MAD2, an essential component in the spindle checkpoint pathway that ensures metaphase is complete before anaphase begins. Combined immunolocalization of MAD2 and a recently cloned maize CENPC homologue indicates that MAD2 localizes to an outer domain of the prometaphase kinetochore. MAD2 staining was primarily observed on mitotic kinetochores that lacked attached microtubules; i.e., at prometaphase or when the microtubules were depolymerized with oryzalin. In contrast, the loss of MAD2 staining in meiosis was not correlated with initial microtubule attachment but was correlated with a measure of tension: the distance between homologous or sister kinetochores (in meiosis I and II, respectively). Further, the tension-sensitive 3F3/2 phosphoepitope colocalized, and was lost concomitantly, with MAD2 staining at the meiotic kinetochore. The mechanism of spindle assembly (discussed here with respect to maize mitosis and meiosis) is likely to affect the relative contributions of attachment and tension. We support the idea that MAD2 is attachment-sensitive and that tension stabilizes microtubule attachments.

Key Words: MAD2 • kinetochore • checkpoint • spindle assembly • meiosis

Abbreviations used in this paper: APC, anaphase-promoting complex; EST, expressed sequence tags; MAP, mitogen-activated protein.

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