Genetics, Vol. 157, 1493-1502, April 2001, Copyright © 2001

The Spindle Checkpoint of the Yeast Saccharomyces cerevisiae Requires Kinetochore Function and Maps to the CBF3 Domain

Richard D. Gardnera, Atasi Poddara, Chris Yellmana, Penny A. Tavorminaa,b, M. Cristina Monteagudoa, and Daniel J. Burkea
a Department of Biochemistry and Molecular Genetics, University of Virginia Medical Center, University of Virginia, Charlottesville, Virginia 22908-0733
b Department of Cell Biology, University of Oklahoma Health Sciences Center, University of Oklahoma, Oklahoma City, Oklahoma 73190

Corresponding author: Daniel J. Burke, Department of Biochemistry and Molecular Genetics, University of Virginia Medical Center, Box 899733, Charlottesville, VA 22908-0733., dburke{at}virginia.edu (E-mail)

Communicating editor: M. LICHTEN

We have measured the activity of the spindle checkpoint in null mutants lacking kinetochore activity in the yeast Saccharomyces cerevisiae. We constructed deletion mutants for nonessential genes by one-step gene replacements. We constructed heterozygous deletions of one copy of essential genes in diploid cells and purified spores containing the deletion allele. In addition, we made gene fusions for three essential genes to target the encoded proteins for proteolysis (degron alleles). We determined that Ndc10p, Ctf13p, and Cep3p are required for checkpoint activity. In contrast, cells lacking Cbf1p, Ctf19p, Mcm21p, Slk19p, Cse4p, Mif2p, Mck1p, and Kar3p are checkpoint proficient. We conclude that the kinetochore plays a critical role in checkpoint signaling in S. cerevisiae. Spindle checkpoint activity maps to a discreet domain within the kinetochore and depends on the CBF3 protein complex.




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