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Vol. 17, Issue 7, 3267-3280, July 2006

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Role of Cell Cycle-regulated Expression in the Localized Incorporation of Cell Wall Proteins in Yeast

Gertien J. Smits^*, Laura R. Schenkman^,, Stanley Brul^*, John R. Pringle^,,, and Frans M. Klis^*

*Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, University of Amsterdam, 1018 WV Amsterdam, The Netherlands; and Department of Biology and Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, NC 27599

Submitted August 9, 2005; Revised April 18, 2006; Accepted April 21, 2006
Monitoring Editor: Anthony Bretscher

The yeast cell wall is an essential organelle that protects the cell from mechanical damage and antimicrobial peptides, participates in cell recognition and adhesion, and is important for the generation and maintenance of normal cell shape. We studied the localization of three covalently bound cell wall proteins in Saccharomyces cerevisiae. Tip1p was found only in mother cells, whereas Cwp2p was incorporated in small-to-medium–sized buds. When the promoter regions of TIP1 and CWP2 (responsible for transcription in early G₁ and S/G₂ phases, respectively) were exchanged, the localization patterns of Tip1p and Cwp2p were reversed, indicating that the localization of cell wall proteins can be completely determined by the timing of transcription during the cell cycle. The third protein, Cwp1p, was incorporated into the birth scar, where it remained for several generations. However, we could not detect any role of Cwp1p in strengthening the birth scar wall or any functional interaction with the proteins that mark the birth scar pole as a potential future budding site. Promoter-exchange experiments showed that expression in S/G₂ phase is necessary but not sufficient for the normal localization of Cwp1p. Studies of mutants in which septum formation is perturbed indicate that the normal asymmetric localization of Cwp1p also depends on the normal timing of septum formation, composition of the septum, or both.

The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

Present address: Department of Genetics, Stanford University Medical Center, Stanford, CA 94305-5120.

Address correspondence to: Gertien Smits ( gertien{at}science.uva.nl)

Abbreviations used: CFW, calcofluor white; DOC, sodium deoxycholate; GFP, green fluorescent protein; GPI, glycosylphosphatidyl-inositol; ORF, open reading frame; TCA, trichloroacetic acid