4.4 Article

Noncore components of the fission yeast γ-tubulin complex

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MOLECULAR BIOLOGY OF THE CELL
卷 17, 期 12, 页码 5075-5093

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AMER SOC CELL BIOLOGY
DOI: 10.1091/mbc.E05-11-1009

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  1. Wellcome Trust Funding Source: Medline

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Relatively little is known about the in vivo function of individual components of the eukaryotic gamma-tubulin complex (gamma-Tuc). We identified three genes, gfh1+, mod21+, and mod22+, in a screen for fission yeast mutants affecting microtubule organization. gfh1+ is a previously characterized gamma-TuC protein weakly similar to human gamma-TuC subunit GCP4, whereas mod21+ is novel and shows weak similarity to human gamma-TuC subunit GCP5. We show that mod21p is a bona fide y-TuC protein and that, like gfh1 Delta mutants, mod21 Delta mutants are viable. We find that gfh1 Delta and mod21A mutants have qualitatively normal microtubule nucleation from all types of microtubule-organizing centers (MTOCs) in vivo but quantitatively reduced nucleation from interphase MTOCs, and this is exacerbated by mutations in mod22+. Simultaneous deletion of gfh1p, mod21p, and alp16p, a third nonessential gamma-TuC protein, does not lead to additive defects, suggesting that all three proteins contribute to a single function. Coimmunoprecipitation experiments suggest that gfh1p and alp16p are codependent for association with a small core gamma-TuC, whereas mod21p is more peripherally associated, and that gfh1p and mod21p may form a subcomplex independently of the small gamma-TuC. Interestingly, sucrose gradient analysis suggests that the major form of the gamma-TuC in fission yeast may be a small complex. We propose that gfh1p, mod21p, and alp16 act as facultative noncore components of the fission yeast gamma-TuC and enhance its microtubule-nucleating ability.

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