In Saccharomyces cerevisiae, Bud8p and Bud9p are homologous plasma membrane glycoproteins that appear to mark the distal and proximal cell poles, respectively, as potential sites for budding in the bipolar pattern. Here we provide evidence that Bud8p is delivered to the presumptive bud site (and thence to the distal pole of the bud) just before bud emergence, and that Bud9p is delivered to the bud side of the mother-bud neck (and thence to the proximal pole of the daughter cell) after activation of the mitotic exit network, just before cytokinesis. Like the delivery of Bud8p, that of Bud9p is actin dependent; unlike the delivery of Bud8p, that of Bud9p is also septin dependent. Interestingly, although the transcription of BUD8 and BUD9 appears to be cell cycle regulated, the abundance of BUD8 mRNA peaks in G2/M and that of BUD9 mRNA peaks in late G1, suggesting that the translation and/or delivery to the cell surface of each protein is delayed and presumably also cell cycle regulated. The importance of time of transcription in localization is supported by promoter-swap experiments: expression of Bud8p from the BUD9 promoter leads to its localization predominantly to the sites typical for Bud9p, and vice versa. Moreover, expression of Bud8p from the BUD9 promoter fails to rescue the budding-pattern defect of a bud8 mutant but fully rescues that of a bud9 mutant. However, although expression of Bud9p from the BUD8 promoter fails to rescue a bud9 mutant, it also rescues only partially the budding-pattern defect of a bud8 mutant, suggesting that some feature(s) of the Bud8p protein is also important for Bud8p function. Experiments with chimeric proteins suggest that the critical element(s) is somewhere in the extracytoplasmic domain of Bud8p.
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4 March 2002
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March 04 2002
The role of cell cycle–regulated expression in the localization of spatial landmark proteins in yeast
Laura R. Schenkman,
Laura R. Schenkman
1Department of Biology, University of North Carolina, Chapel Hill, NC 27599
2Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, NC 27599
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Carlo Caruso,
Carlo Caruso
1Department of Biology, University of North Carolina, Chapel Hill, NC 27599
2Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, NC 27599
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Nicolas Pagé,
Nicolas Pagé
3Department of Biology, McGill University, Montreal H3A 1B1, Canada
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John R. Pringle
John R. Pringle
1Department of Biology, University of North Carolina, Chapel Hill, NC 27599
2Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, NC 27599
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Laura R. Schenkman
1Department of Biology, University of North Carolina, Chapel Hill, NC 27599
2Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, NC 27599
Carlo Caruso
1Department of Biology, University of North Carolina, Chapel Hill, NC 27599
2Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, NC 27599
Nicolas Pagé
3Department of Biology, McGill University, Montreal H3A 1B1, Canada
John R. Pringle
1Department of Biology, University of North Carolina, Chapel Hill, NC 27599
2Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, NC 27599
Address correspondence to John R. Pringle, Dept. of Biology, CB #3280, 421 Fordham Hall, University of North Carolina, Chapel Hill, NC 27599. Tel.: (919) 962-2293. Fax: (919) 962-0320. E-mail: [email protected]
The online version of this article contains supplemental material.
N. Pagé's present address is Swiss Institute for Experimental Cancer Research, 1066 Epalinges/VD, Switzerland.
*
Abbreviations used in this paper: GFP, green fluorescent protein; Lat A, Latrunculin A.
Received:
July 10 2001
Revision Received:
January 18 2002
Accepted:
January 23 2002
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2002
J Cell Biol (2002) 156 (5): 829–841.
Article history
Received:
July 10 2001
Revision Received:
January 18 2002
Accepted:
January 23 2002
Citation
Laura R. Schenkman, Carlo Caruso, Nicolas Pagé, John R. Pringle; The role of cell cycle–regulated expression in the localization of spatial landmark proteins in yeast . J Cell Biol 4 March 2002; 156 (5): 829–841. doi: https://doi.org/10.1083/jcb.200107041
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