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Molecular and Cellular Biology, April 1999, p. 2535-2546, Vol. 19, No. 4
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The Schizosaccharomyces pombe dim1+ Gene Interacts with the Anaphase-Promoting Complex or Cyclosome (APC/C) Component lid1+ and Is Required for APC/C Function

Lynne D. Berry,1 Anna Feoktistova,2 Melanie D. Wright,1 and Kathleen L. Gould1,2,*

Howard Hughes Medical Institute2 and Department of Cell Biology,1 Vanderbilt University School of Medicine, Nashville, Tennessee 37232

Received 5 August 1998/Returned for modification 21 September 1998/Accepted 22 December 1998

The Schizosaccharomyces pombe dim1+ gene is required for entry into mitosis and for chromosome segregation during mitosis. To further understand dim1p function, we undertook a synthetic lethal screen with the temperature-sensitive dim1-35 mutant and isolated lid (for lethal in dim1-35) mutants. Here, we describe the temperature-sensitive lid1-6 mutant. At the restrictive temperature of 36°C, lid1-6 mutant cells arrest with a "cut" phenotype similar to that of cut4 and cut9 mutants. An epitope-tagged version of lid1p is a component of a multiprotein ~20S complex; the presence of lid1p in this complex depends upon functional cut9+. lid1p-myc coimmunoprecipitates with several other proteins, including cut9p and nuc2p, and the presence of cut9p in a 20S complex depends upon the activity of lid1+. Further, lid1+ function is required for the multiubiquitination of cut2p, an anaphase-promoting complex or cyclosome (APC/C) target. Thus, lid1p is a component of the S. pombe APC/C. In dim1 mutants, the abundances of lid1p and the APC/C complex decline significantly, and the ubiquitination of an APC/C target is abolished. These data suggest that at least one role of dim1p is to maintain or establish the steady-state level of the APC/C.

* Corresponding author. Mailing address: HHMI and Dept. of Cell Biology, Vanderbilt University School of Medicine, Nashville, TN 37232. Phone: (615) 343-9502. Fax: (615) 343-0723. E-mail: kathy.gould{at}mcmail.vanderbilt.edu.

Molecular and Cellular Biology, April 1999, p. 2535-2546, Vol. 19, No. 4
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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