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Fission yeast Scp3 potentially maintains microtubule orientation through bundling.

Ozaki K, Chikashige Y, Hiraoka Y, Matsumoto T - PLoS ONE (2015)

Bottom Line: In this study, we investigated the function of Scp3 along with the effect of CIPC in the fission yeast Schizosaccharomyces pombe.Functional analysis suggested that Scp3 functions independently from Ase1, a protein largely required for the bundling of the mitotic spindle.These results suggested that multiple systems are independently involved to ensure microtubule orientation by bundling in fission yeast.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Biostudies, Kyoto University, Kyoto, Kyoto, Japan.

ABSTRACT
Microtubules play important roles in organelle transport, the maintenance of cell polarity and chromosome segregation and generally form bundles during these processes. The fission yeast gene scp3+ was identified as a multicopy suppressor of the cps3-81 mutant, which is hypersensitive to isopropyl N-3-chlorophenylcarbamate (CIPC), a poison that induces abnormal multipolar spindle formation in higher eukaryotes. In this study, we investigated the function of Scp3 along with the effect of CIPC in the fission yeast Schizosaccharomyces pombe. Microscopic observation revealed that treatment with CIPC, cps3-81 mutation and scp3+ gene deletion disturbed the orientation of microtubules in interphase cells. Overexpression of scp3+ suppressed the abnormal orientation of microtubules by promoting bundling. Functional analysis suggested that Scp3 functions independently from Ase1, a protein largely required for the bundling of the mitotic spindle. A strain lacking the ase1+ gene was more sensitive to CIPC, with the drug affecting the integrity of the mitotic spindle, indicating that CIPC has a mitotic target that has a role redundant with Ase1. These results suggested that multiple systems are independently involved to ensure microtubule orientation by bundling in fission yeast.

No MeSH data available.


Related in: MedlinePlus

Abnormal microtubule orientation in Δscp3.(A) The ∆scp3 strain expressing GFP-Atb2 and Sad1-mCherry from the native promoters was treated with DMSO or 260 μM CIPC for 5 hours in EMM medium at 30°C. The bar indicates 5 μm. (B) Microtubule orientation was analyzed as in Fig. 2B. (C) Nonparametric Mann-Whitney U test of (B). The red lines are the median.
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pone.0120109.g005: Abnormal microtubule orientation in Δscp3.(A) The ∆scp3 strain expressing GFP-Atb2 and Sad1-mCherry from the native promoters was treated with DMSO or 260 μM CIPC for 5 hours in EMM medium at 30°C. The bar indicates 5 μm. (B) Microtubule orientation was analyzed as in Fig. 2B. (C) Nonparametric Mann-Whitney U test of (B). The red lines are the median.

Mentions: The scp3+ gene was replaced with the ura4+ gene in a diploid strain, and a stable Ura4+ diploid (scp3+::ura4+/+) was examined by tetrad analysis. Most of the tetrads produced four viable spores in which the Ura4 marker segregated into 2:2, indicating that the scp3+ gene is not essential for viability. Observation of MTs in a strain deleted for the scp3+ gene (Δscp3) revealed the misorientation of MTs (Fig. 5A). As shown in Fig. 5B and 5C, the drug did not cause any additional effects on the orientation of MT in the Δscp3 strain, suggesting that the drug might target Scp3.


Fission yeast Scp3 potentially maintains microtubule orientation through bundling.

Ozaki K, Chikashige Y, Hiraoka Y, Matsumoto T - PLoS ONE (2015)

Abnormal microtubule orientation in Δscp3.(A) The ∆scp3 strain expressing GFP-Atb2 and Sad1-mCherry from the native promoters was treated with DMSO or 260 μM CIPC for 5 hours in EMM medium at 30°C. The bar indicates 5 μm. (B) Microtubule orientation was analyzed as in Fig. 2B. (C) Nonparametric Mann-Whitney U test of (B). The red lines are the median.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4359140&req=5

pone.0120109.g005: Abnormal microtubule orientation in Δscp3.(A) The ∆scp3 strain expressing GFP-Atb2 and Sad1-mCherry from the native promoters was treated with DMSO or 260 μM CIPC for 5 hours in EMM medium at 30°C. The bar indicates 5 μm. (B) Microtubule orientation was analyzed as in Fig. 2B. (C) Nonparametric Mann-Whitney U test of (B). The red lines are the median.
Mentions: The scp3+ gene was replaced with the ura4+ gene in a diploid strain, and a stable Ura4+ diploid (scp3+::ura4+/+) was examined by tetrad analysis. Most of the tetrads produced four viable spores in which the Ura4 marker segregated into 2:2, indicating that the scp3+ gene is not essential for viability. Observation of MTs in a strain deleted for the scp3+ gene (Δscp3) revealed the misorientation of MTs (Fig. 5A). As shown in Fig. 5B and 5C, the drug did not cause any additional effects on the orientation of MT in the Δscp3 strain, suggesting that the drug might target Scp3.

Bottom Line: In this study, we investigated the function of Scp3 along with the effect of CIPC in the fission yeast Schizosaccharomyces pombe.Functional analysis suggested that Scp3 functions independently from Ase1, a protein largely required for the bundling of the mitotic spindle.These results suggested that multiple systems are independently involved to ensure microtubule orientation by bundling in fission yeast.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Biostudies, Kyoto University, Kyoto, Kyoto, Japan.

ABSTRACT
Microtubules play important roles in organelle transport, the maintenance of cell polarity and chromosome segregation and generally form bundles during these processes. The fission yeast gene scp3+ was identified as a multicopy suppressor of the cps3-81 mutant, which is hypersensitive to isopropyl N-3-chlorophenylcarbamate (CIPC), a poison that induces abnormal multipolar spindle formation in higher eukaryotes. In this study, we investigated the function of Scp3 along with the effect of CIPC in the fission yeast Schizosaccharomyces pombe. Microscopic observation revealed that treatment with CIPC, cps3-81 mutation and scp3+ gene deletion disturbed the orientation of microtubules in interphase cells. Overexpression of scp3+ suppressed the abnormal orientation of microtubules by promoting bundling. Functional analysis suggested that Scp3 functions independently from Ase1, a protein largely required for the bundling of the mitotic spindle. A strain lacking the ase1+ gene was more sensitive to CIPC, with the drug affecting the integrity of the mitotic spindle, indicating that CIPC has a mitotic target that has a role redundant with Ase1. These results suggested that multiple systems are independently involved to ensure microtubule orientation by bundling in fission yeast.

No MeSH data available.


Related in: MedlinePlus