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PMID: 15356260 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Interactions among Rax1p, Rax2p, Bud8p, and Bud9p in marking cortical sites for bipolar bud-site selection in yeast.

Molecular biology of the cell ·Vol. 15 ·No. 11 ·2004-11-00 ·Pages 5145-57

Kang PJ, Angerman E, Nakashima K, Pringle JR, Park HO

Abstract

In the budding yeast Saccharomyces cerevisiae, selection of the bud site determines the axis of polarized cell growth and eventual oriented cell division. Bud sites are selected in specific patterns depending on cell type. These patterns appear to depend on distinct types of marker proteins in the cell cortex; in particular, the bipolar budding of diploid cells depends on persistent landmarks at the birth-scar-distal and -proximal poles that involve the proteins Bud8p and Bud9p, respectively. Rax1p and Rax2p also appear to function specifically in bipolar budding, and we report here a further characterization of these proteins and of their interactions with Bud8p and Bud9p. Rax1p and Rax2p both appear to be integral membrane proteins. Although commonly used programs predict different topologies for Rax2p, glycosylation studies indicate that it has a type I orientation, with its long N-terminal domain in the extracytoplasmic space. Analysis of rax1 and rax2 mutant budding patterns indicates that both proteins are involved in selecting bud sites at both the distal and proximal poles of daughter cells as well as near previously used division sites on mother cells. Consistent with this, GFP-tagged Rax1p and Rax2p were both observed at the distal pole as well as at the division site on both mother and daughter cells; localization to the division sites was persistent through multiple cell cycles. Localization of Rax1p and Rax2p was interdependent, and biochemical studies showed that these proteins could be copurified from yeast. Bud8p and Bud9p could also be copurified with Rax1p, and localization studies provided further evidence of interactions. Localization of Rax1p and Rax2p to the bud tip and distal pole depended on Bud8p, and normal localization of Bud8p was partially dependent on Rax1p and Rax2p. Although localization of Rax1p and Rax2p to the division site did not appear to depend on Bud9p, normal localization of Bud9p appeared largely or entirely dependent on Rax1p and Rax2p. Taken together, the results indicate that Rax1p and Rax2p interact closely with each other and with Bud8p and Bud9p in the establishment and/or maintenance of the cortical landmarks for bipolar budding.

MeSH Terms
Binding Sites Cell Division Cell Membrane/metabolism Cell Polarity Cell Proliferation Cloning, Molecular Cytoplasm/metabolism Fungal Proteins/physiology Gene Deletion Glycoproteins/metabolism Green Fluorescent Proteins/metabolism Membrane Glycoproteins/metabolism Membrane Proteins Mutation Plasmids/metabolism Protein Binding Protein Structure, Tertiary Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins/metabolism,physiology
Chemicals
BUD8 protein, S cerevisiae BUD9 protein, S cerevisiae Fungal Proteins Glycoproteins Membrane Glycoproteins Membrane Proteins RAX1 protein, S cerevisiae Rax2 protein, S cerevisiae Saccharomyces cerevisiae Proteins Green Fluorescent Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Kang Pil Jung
Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210-1292, USA.
Angerman Elizabeth
Nakashima Kenichi
Pringle John R
Park Hay-Oak
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e04-07-0600
Published
2004-11-00
Epub
2004-00-08
Pages
5145-57
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC524791
Subset
IM
Grants
NIGMS NIH HHS · GM56997 · United States
NIGMS NIH HHS · R37 GM031006 · United States
NIGMS NIH HHS · GM31006 · United States
NIGMS NIH HHS · R01 GM056997 · United States
NIGMS NIH HHS · R01 GM031006 · United States
NIGMS NIH HHS · R01 GM056997-05 · United States
External Links
PubMed source DOI: 10.1091/mbc.e04-07-0600 PMC Full Text View journal details
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