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MBC in Press, published online ahead of print May 14, 2008
Mol. Biol. Cell 10.1091/mbc.E08-02-0148

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Submitted on February 13, 2008
Revised on May 2, 2008
Accepted on May 7, 2008

Intrinsic and CDK-dependent Control of Spindle Pole Body Duplication in Budding Yeast

Laura A. Simmons Kovacs,* Christine L. Nelson,{dagger} and Steven B. Haase*

*Department of Biology, Duke University, Durham, NC 27708-0338; {dagger}Biochemistry Department, Duke University Medical Center, Durham, NC 27710

Monitoring Editor: Kerry S. Bloom

Centrosome duplication must be tightly controlled so that duplication occurs only once each cell cycle. Accumulation of multiple centrosomes can result in the assembly of a multi-polar spindle and lead to chromosome mis-segregation and genomic instability. In metazoans, a centrosome-intrinsic mechanism prevents reduplication until centriole disengagement. Mitotic cyclin/CDKs prevent reduplication of the budding yeast centrosome, called a spindle pole body (SPB) in late S phase and G2/M, but the mechanism remains unclear. How SPB reduplication is prevented early in the cell cycle is also not understood. Here we show that, similar to metazoans, an SPB-intrinsic mechanism prevents reduplication early in the cell cycle. We also show that mitotic cyclins can inhibit SPB duplication when expressed before satellite assembly in early G1, but not later in G1, after the satellite had assembled. Moreover, electron microscopy revealed that SPBs do not assemble a satellite in cells expressing Clb2 in early G1. Finally, we demonstrate that Clb2 must localize to the cytoplasm in order to inhibit SPB duplication, suggesting the possibility for direct CDK inhibition of satellite components. These two mechanisms, intrinsic and extrinsic control by CDK, evoke two-step system that prevents SPB reduplication throughout the cell cycle.

Address correspondence to: Steven B. Haase (shaase{at}duke.edu)






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