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PMID: 10982392 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Bim1p/Yeb1p mediates the Kar9p-dependent cortical attachment of cytoplasmic microtubules.

Molecular biology of the cell ·Vol. 11 ·No. 9 ·2000-09-00 ·Pages 2949-59

Miller RK, Cheng SC, Rose MD

Abstract

In Saccharomyces cerevisiae, positioning of the mitotic spindle depends on the interaction of cytoplasmic microtubules with the cell cortex. In this process, cortical Kar9p in the bud acts as a link between the actin and microtubule cytoskeletons. To identify Kar9p-interacting proteins, a two-hybrid screen was conducted with the use of full-length Kar9p as bait, and three genes were identified: BIM1, STU2, and KAR9 itself. STU2 encodes a component of the spindle pole body. Bim1p is the yeast homologue of the human microtubule-binding protein EB1, which is a binding partner to the adenomatous polyposis coli protein involved in colon cancer. Eighty-nine amino acids within the third quarter of Bim1p was sufficient to confer interaction with Kar9p. The two-hybrid interactions were confirmed with the use of coimmunoprecipitation experiments. Genetic analysis placed Bim1p in the Kar9p pathway for nuclear migration. Bim1p was not required for Kar9p's cortical or spindle pole body localization. However, deletion of BIM1 eliminated Kar9p localization along cytoplasmic microtubules. Furthermore, in the bim1 mutants, the cytoplasmic microtubules no longer intersected the cortical dot of Green Fluorescent Protein-Kar9p. These experiments demonstrate that the interaction of cytoplasmic microtubules with the Kar9p cortical attachment site requires the microtubule-binding protein Bim1p.

MeSH Terms
Amino Acid Sequence Animals Binding Sites Cell Cycle Proteins/chemistry,genetics,metabolism Cytoplasm/physiology,ultrastructure Fungal Proteins/chemistry,genetics,metabolism Genes, Fungal Genotype Humans Microtubule Proteins/chemistry,genetics,metabolism Microtubule-Associated Proteins/chemistry,genetics,metabolism Microtubules/physiology,ultrastructure Molecular Sequence Data Nuclear Proteins/genetics,metabolism Recombinant Proteins/chemistry,metabolism Saccharomyces cerevisiae/genetics,physiology,ultrastructure Saccharomyces cerevisiae Proteins Sequence Alignment Sequence Homology, Amino Acid
Chemicals
BIM1 protein, S cerevisiae Cell Cycle Proteins EB1 microtubule binding proteins Fungal Proteins KAR9 protein, S cerevisiae Microtubule Proteins Microtubule-Associated Proteins Nuclear Proteins Recombinant Proteins STU2 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Miller R K
Department of Molecular Biology, Lewis Thomas Laboratory, Princeton University, Princeton, New Jersey 08544, USA.
Cheng S C
Rose M D
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.11.9.2949
Published
2000-09-00
Pages
2949-59
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC14967
Subset
IM
Grants
NIGMS NIH HHS · R01 GM037739 · United States
NIGMS NIH HHS · GM37739 · United States
External Links
PubMed source DOI: 10.1091/mbc.11.9.2949 PMC Full Text View journal details
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