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J. Cell Biol.,
Volume 141, Number 5, June 1, 1998 1193-1205
* Department of Cell Biology, University of Virginia, Charlottesville, Virginia 22908; In yeast, the Mad2 protein is required for the
M phase arrest induced by microtubule inhibitors, but
the protein is not essential under normal culture conditions. We tested whether the Mad2 protein participates
in regulating the timing of anaphase onset in mammalian cells in the absence of microtubule drugs. When
microinjected into living prophase or prometaphase
PtK1 cells, anti-Mad2 antibody induced the onset of
anaphase prematurely during prometaphase, before the
chromosomes had assembled at the metaphase plate.
Anti-Mad2 antibody-injected cells completed all aspects of anaphase including chromatid movement to
the spindle poles and pole-pole separation. Identical
results were obtained when primary human keratinocytes were injected with anti-Mad2 antibody. These
studies suggest that Mad2 protein function is essential
for the timing of anaphase onset in somatic cells at each
mitosis. Thus, in mammalian somatic cells, the spindle
checkpoint appears to be a component of the timing
mechanism for normal mitosis, blocking anaphase onset until all chromosomes are aligned at the metaphase
plate.
Section of Biochemistry, Molecular and
Cell Biology, Cornell University, Ithaca, New York 14853; and § Department of Physiology, University of California,
San Francisco, California 94143
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