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Eukaryotic Cell, June 2005, p. 1018-1028, Vol. 4, No. 6
1535-9778/05/$08.00+0     doi:10.1128/EC.4.6.1018-1028.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

ACE2, CBK1, and BUD4 in Budding and Cell Separation

Warren P. Voth, Aileen E. Olsen, Mohammed Sbia,{dagger} Karen H. Freedman,{ddagger} and David J. Stillman*

Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, Utah 84132

Received 14 January 2005/ Accepted 6 April 2005

Mutations in the RAM network genes, including CBK1, MOB2, KIC1, HYM1, and TAO3, cause defects in bud site selection, asymmetric apical growth, and mating projections. Additionally, these mutants show altered colony morphology, cell separation defects, and reduced CTS1 expression, phenotypes also seen by mutating the Ace2 transcription factor. We show that an ACE2 multicopy plasmid suppresses the latter three defects of RAM network mutations, demonstrating that Ace2 is downstream of the RAM network and suggesting that these phenotypes are caused by reduced expression of Ace2 target genes. We show that wild-type W303 strains have a bud4 mutation and that combining bud4 with either ace2 or cbk1 in haploids results in altered colony morphology. We describe a timed sedimentation assay that allows quantitation of cytokinesis defects and subtle changes in budding pattern and cell shape. Experiments examining budding patterns and sedimentation rates both show that Ace2 and Cbk1 have independent functions in addition to their common pathway in transcription of genes such as CTS1. SWI5 encodes a transcription factor paralogous to ACE2. Additive effects are seen in cbk1 swi5 strains, and we show that activation of some target genes, such as EGT2, requires either Swi5 or Ace2 with Cbk1. The relative roles and interactions of Ace2, Cbk1, and Bud4 in bud site selection, polarized growth, and cell separation are discussed.

* Corresponding author. Mailing address: Department of Pathology, University of Utah, 30 North 1900 East, Salt Lake City, UT 84132-2501. Phone: (801) 581-5429. Fax: (801) 581-4517. E-mail: david.stillman{at}path.utah.edu.

{dagger} Present address: Department of Oncological Sciences, University of Utah, Salt Lake City, UT 84132.

{ddagger} Present address: University of Wisconsin, Madison, WI 53706.

Eukaryotic Cell, June 2005, p. 1018-1028, Vol. 4, No. 6
1535-9778/05/$08.00+0     doi:10.1128/EC.4.6.1018-1028.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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