Modulation of Alp4 function in Schizosaccharomyces pombe induces novel phenotypes that imply distinct functions for nuclear and cytoplasmic gamma-tubulin complexes.
The gamma-tubulin complex acts as a nucleation unit for microtubule assembly. It remains unknown, however, how spatial and temporal regulation of the complex activity affects microtubule-mediated cellular processes. Alp4 is one of the essential components of the S. pombe gamma-tubulin complex. We show here that overproduction of a carboxy-terminal form of Alp4 (Alp4C) and its derivatives tagged to a nuclear localization signal or to a nuclear export signal affect localization of gamma-tubulin complexes and induces novel phenotypes that reflect distinct functions of nuclear and cytoplasmic gamma-tubulin complexes. Nuclear Alp4C induces a Wee1-dependent G2 delay, reduces the levels of the gamma-tubulin complex at the spindle pole body, and results in defects in mitotic progression including spindle assembly, cytoplasmic microtubule disassembly, and chromosome segregation. In contrast, cytoplasmic Alp4C induces oscillatory nuclear movement and affects levels of cell polarity markers, Bud6 and Tip1, at the cell ends. These results demonstrate that regulation of nuclear gamma-tubulin complex activity is essential for cell cycle progression through the G2/M boundary and M phase, whereas regulation of cytoplasmic gamma-tubulin complex activity is important for nuclear positioning and cell polarity control during interphase.
Masuda
Hirohisa
H
Cell Biology Group and CREST/JST, Kansai Advanced Research Center, National Institute of Information and Communications Technology, Kobe, 651-2492, Japan. hmasuda@nict.go.jp
Toda
Takashi
T
Miyamoto
Rumi
R
Haraguchi
Tokuko
T
Hiraoka
Yasushi
Y
D018797:Cell Cycle Proteins / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism*; D016195:G2 Phase / Q000502:physiology; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000502:physiology*; D008870:Microtubules / Q000502:physiology; D009154:Mutation; D019913:Nuclear Localization Signals / Q000502:physiology; D009687:Nuclear Proteins / Q000502:physiology; D010641:Phenotype; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D012568:Schizosaccharomyces / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D029702:Schizosaccharomyces pombe Proteins / Q000096:biosynthesis / Q000502:physiology*; D008941:Spindle Apparatus / Q000502:physiology; D013696:Temperature; D014404:Tubulin / Q000502:physiology*
C512340:Alp4 protein, S pombe; C512341:Alp6 protein, S pombe; D018797:Cell Cycle Proteins; D008869:Microtubule-Associated Proteins; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D029702:Schizosaccharomyces pombe Proteins; D014404:Tubulin; C068689:wee1 protein, S pombe; D011505:Protein-Tyrosine Kinases