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PMID: 15492045 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Differential contribution of Bud6p and Kar9p to microtubule capture and spindle orientation in S. cerevisiae.

The Journal of cell biology ·Vol. 167 ·No. 2 ·2004-10-25 ·Pages 231-44

Huisman SM, Bales OA, Bertrand M, Smeets MF, Reed SI, Segal M

Abstract

In Saccharomyces cerevisiae, spindle orientation is controlled by a temporal and spatial program of microtubule (MT)-cortex interactions. This program requires Bud6p/Aip3p to direct the old pole to the bud and confine the new pole to the mother cell. Bud6p function has been linked to Kar9p, a protein guiding MTs along actin cables. Here, we show that Kar9p does not mediate Bud6p functions in spindle orientation. Based on live microscopy analysis, kar9Delta cells maintained Bud6p-dependent MT capture. Conversely, bud6Delta cells supported Kar9p-associated MT delivery to the bud. Moreover, additive phenotypes in bud6Delta kar9Delta or bud6Delta dyn1Delta mutants underscored the separate contributions of Bud6p, Kar9p, and dynein to spindle positioning. Finally, tub2C354S, a mutation decreasing MT dynamics, suppressed a kar9Delta mutation in a BUD6-dependent manner. Thus, Kar9p-independent capture at Bud6p sites can effect spindle orientation provided MT turnover is reduced. Together, these results demonstrate Bud6p function in MT capture at the cell cortex, independent of Kar9p-mediated MT delivery along actin cables.

MeSH Terms
Actins/chemistry,metabolism Cytoskeleton/metabolism Dyneins/physiology Fungal Proteins/chemistry Green Fluorescent Proteins/metabolism Microfilament Proteins/physiology Microscopy, Fluorescence Microtubule Proteins/chemistry Microtubules/ultrastructure Models, Biological Mutation Nuclear Proteins/physiology Phenotype Plasmids/metabolism Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins/physiology Spindle Apparatus Time Factors
Chemicals
Actins BUD6 protein, S cerevisiae Fungal Proteins KAR9 protein, S cerevisiae Microfilament Proteins Microtubule Proteins Nuclear Proteins Saccharomyces cerevisiae Proteins Green Fluorescent Proteins Dyneins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Huisman Stephen M
Department of Genetics, University of Cambridge, Cambridge, CB2 3EH UK.
Bales Olivia A M
Bertrand Marie
Smeets Monique F M A
Reed Steven I
Segal Marisa
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200407167
Published
2004-10-25
Epub
2004-00-18
Pages
231-44
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2172562
Subset
IM
Grants
Wellcome Trust · United Kingdom
NIGMS NIH HHS · R01 GM038328 · United States
NIGMS NIH HHS · R37 GM038328 · United States
External Links
PubMed source DOI: 10.1083/jcb.200407167 PMC Full Text View journal details
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