VINTAGE ANTIQUE JEWELRY

CENP-A exceeds microtubule attachment sites in centromere clusters of both budding and fission yeast (.jcb)

The stoichiometries of kinetochores and their constituent proteins in yeast
and vertebrate cells were determined using the histone H3 variant CENP-A,
known as Cse4 in budding yeast, as a counting standard. One Cse4-containing
nucleosome exists in the centromere (CEN) of each chromosome, so it has been
assumed that each anaphase CEN/kinetochore cluster contains 32 Cse4 molecules.
We report that anaphase CEN clusters instead contained approximately fourfold
more Cse4 in _Saccharomyces cerevisiae_ and ~40-fold more CENP-A (Cnp1) in
_Schizosaccharomyces pombe_ than predicted. These results suggest that the
number of CENP-A molecules exceeds the number of kinetochore-microtubule (MT)
attachment sites on each chromosome and that CENP-A is not the sole
determinant of kinetochore assembly sites in either yeast. In addition, we
show that fission yeast has enough Dam1-DASH complex for ring formation around
attached MTs. The results of this study suggest the need for significant
revision of existing CEN/kinetochore architectural models.

.jcb

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