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Fission yeast mto2p regulates microtubule nucleation by the centrosomin-related protein mto1p.
- Source :
-
Molecular biology of the cell [Mol Biol Cell] 2005 Jun; Vol. 16 (6), pp. 3040-51. Date of Electronic Publication: 2005 Jan 19. - Publication Year :
- 2005
-
Abstract
- From an insertional mutagenesis screen, we isolated a novel gene, mto2+, involved in microtubule organization in fission yeast. mto2Delta strains are viable but exhibit defects in interphase microtubule nucleation and in formation of the postanaphase microtubule array at the end of mitosis. The mto2Delta defects represent a subset of the defects displayed by cells deleted for mto1+ (also known as mod20+ and mbo1+), a centrosomin-related protein required to recruit the gamma-tubulin complex to cytoplasmic microtubule-organizing centers (MTOCs). We show that mto2p colocalizes with mto1p at MTOCs throughout the cell cycle and that mto1p and mto2p coimmunoprecipitate from cytoplasmic extracts. In vitro studies suggest that mto2p binds directly to mto1p. In mto2Delta mutants, although some aspects of mto1p localization are perturbed, mto1p can still localize to spindle pole bodies and the cell division site and to "satellite" particles on interphase microtubules. In mto1Delta mutants, localization of mto2p to all of these MTOCs is strongly reduced or absent. We also find that in mto2Delta mutants, cytoplasmic forms of the gamma-tubulin complex are mislocalized, and the gamma-tubulin complex no longer coimmunoprecipitates with mto1p from cell extracts. These experiments establish mto2p as a major regulator of mto1p-mediated microtubule nucleation by the gamma-tubulin complex.
- Subjects :
- Antibodies, Monoclonal metabolism
Cell Cycle
Fluorescent Antibody Technique
Glutathione Transferase metabolism
Green Fluorescent Proteins metabolism
Microscopy, Video
Microtubule-Organizing Center metabolism
Microtubules metabolism
Mutagenesis, Insertional
Mutation
Precipitin Tests
Protein Binding
Recombinant Proteins metabolism
Schizosaccharomyces cytology
Schizosaccharomyces genetics
Schizosaccharomyces growth & development
Schizosaccharomyces metabolism
Spindle Apparatus metabolism
Tubulin metabolism
Fungal Proteins genetics
Fungal Proteins metabolism
Gene Expression Regulation, Fungal
Genes, Fungal
Microtubule-Associated Proteins genetics
Microtubule-Associated Proteins metabolism
Schizosaccharomyces pombe Proteins genetics
Schizosaccharomyces pombe Proteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1059-1524
- Volume :
- 16
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Molecular biology of the cell
- Publication Type :
- Academic Journal
- Accession number :
- 15659644
- Full Text :
- https://doi.org/10.1091/mbc.e04-11-1003