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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

Microtubules in cps3–81 treated with CIPC.The cps3–81 mutant expressing GFP-Atb2 and Sad1-mCherry, as in Fig. 1A, was treated with DMSO (control) or 260 μM CIPC for 5 hours in EMM medium at 30°C. The bar indicates 5 μm.
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pone.0120109.g003: Microtubules in cps3–81 treated with CIPC.The cps3–81 mutant expressing GFP-Atb2 and Sad1-mCherry, as in Fig. 1A, was treated with DMSO (control) or 260 μM CIPC for 5 hours in EMM medium at 30°C. The bar indicates 5 μm.

Mentions: The cps3–81 mutant which is hypersensitive to CIPC, cannot grow in the presence of 260 μM of the drug [34,46]. Although the cps3+ gene was genetically mapped very closed to the arg1 locus (chromosome III), the gene has not been cloned to date [34]. As shown in Figs. 2B and 3, the MTs in the mutant were misoriented even when the cells were grown in the absence of CIPC. In the medium containing the drug at a lethal concentration (260 μM), the cps3–81 mutant cells exhibited abnormal MT morphology that was slightly more prominent than that observed in the cells grown in the absence of the drug (Figs. 2B and 3). Our examination of the morphology of MTs in the cells failed to demonstrate why the increase in the dose of CIPC kills the cells. Nonetheless, it did indicate that CIPC largely affects the orientation of MTs during interphase in fission yeast.


Fission yeast Scp3 potentially maintains microtubule orientation through bundling.

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

Microtubules in cps3–81 treated with CIPC.The cps3–81 mutant expressing GFP-Atb2 and Sad1-mCherry, as in Fig. 1A, was treated with DMSO (control) or 260 μM CIPC for 5 hours in EMM medium at 30°C. The bar indicates 5 μm.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0120109.g003: Microtubules in cps3–81 treated with CIPC.The cps3–81 mutant expressing GFP-Atb2 and Sad1-mCherry, as in Fig. 1A, was treated with DMSO (control) or 260 μM CIPC for 5 hours in EMM medium at 30°C. The bar indicates 5 μm.
Mentions: The cps3–81 mutant which is hypersensitive to CIPC, cannot grow in the presence of 260 μM of the drug [34,46]. Although the cps3+ gene was genetically mapped very closed to the arg1 locus (chromosome III), the gene has not been cloned to date [34]. As shown in Figs. 2B and 3, the MTs in the mutant were misoriented even when the cells were grown in the absence of CIPC. In the medium containing the drug at a lethal concentration (260 μM), the cps3–81 mutant cells exhibited abnormal MT morphology that was slightly more prominent than that observed in the cells grown in the absence of the drug (Figs. 2B and 3). Our examination of the morphology of MTs in the cells failed to demonstrate why the increase in the dose of CIPC kills the cells. Nonetheless, it did indicate that CIPC largely affects the orientation of MTs during interphase in fission yeast.

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