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Regulation of Rho-GEF Rgf3 by the arrestin Art1 in fission yeast cytokinesis.

Davidson R, Laporte D, Wu JQ - Mol. Biol. Cell (2014)

Bottom Line: Using an Rgf3 conditional mutant and mislocalization experiments, we found that Art1 and Rgf3 are interdependent for localization to the division site.As expected, active Rho1 levels at the division site are reduced in art1∆ and rgf3 mutant cells.Taken together, these data reveal that the arrestin family protein Art1 regulates the protein levels and localization of the Rho-GEF Rgf3, which in turn modulates active Rho1 levels during fission yeast cytokinesis.

View Article: PubMed Central - PubMed

Affiliation: Graduate Program of Molecular, Cellular, and Developmental Biology, The Ohio State University, Columbus, OH 43210 Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210.

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Art1 affects Rgf3 localization and protein levels. (A and B) Micrographs (A) and quantifications (B) of Rgf3-mECitrine in wt (JW2748), art1∆ (JW5232), 3nmt1-art1 (JW4889), and Art1-mECitrine (JW2694). (A) Arrows mark septa and arrowheads mark the contractile ring. (B) The numbers of Art1 and Rgf3 molecules globally in the whole cell and locally in mature but full-sized contractile rings. (C) Art1 affects Rgf3 protein levels. Anti-Myc antibody was used to detect Rgf3-13Myc (asterisks) in cell extracts from rgf3-13Myc (wt, JW2421) and rgf3-13Myc art1∆ (JW4816) strains grown in YE5S (top), or from rgf3-13Myc (wt, JW2421) and rgf3-13Myc 3nmt1-art1 (JW5129) strains induced in EMM5S for 24 h (bottom). The relative ratios of Rgf3 protein levels are given (n = 2). Scale bar: 5 μm.
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Figure 4: Art1 affects Rgf3 localization and protein levels. (A and B) Micrographs (A) and quantifications (B) of Rgf3-mECitrine in wt (JW2748), art1∆ (JW5232), 3nmt1-art1 (JW4889), and Art1-mECitrine (JW2694). (A) Arrows mark septa and arrowheads mark the contractile ring. (B) The numbers of Art1 and Rgf3 molecules globally in the whole cell and locally in mature but full-sized contractile rings. (C) Art1 affects Rgf3 protein levels. Anti-Myc antibody was used to detect Rgf3-13Myc (asterisks) in cell extracts from rgf3-13Myc (wt, JW2421) and rgf3-13Myc art1∆ (JW4816) strains grown in YE5S (top), or from rgf3-13Myc (wt, JW2421) and rgf3-13Myc 3nmt1-art1 (JW5129) strains induced in EMM5S for 24 h (bottom). The relative ratios of Rgf3 protein levels are given (n = 2). Scale bar: 5 μm.

Mentions: Because Art1 and Rgf3 interact with each other and Rgf3 plays a role in Art1 localization, it is interesting to know whether Art1 is involved in Rgf3 localization. In art1∆ cells, Rgf3 localization to the contractile ring was essentially abolished, and the intensity of Rgf3 in the cytoplasm was also reduced compared with wt (Figure 4A). However, a weak Rgf3 signal at the division site was detected in cells with a complete septum (Figure 4A, arrows). In contrast, Rgf3 intensities in the cytoplasm and the division site obviously increased when Art1 was overexpressed (Figure 4A, right).


Regulation of Rho-GEF Rgf3 by the arrestin Art1 in fission yeast cytokinesis.

Davidson R, Laporte D, Wu JQ - Mol. Biol. Cell (2014)

Art1 affects Rgf3 localization and protein levels. (A and B) Micrographs (A) and quantifications (B) of Rgf3-mECitrine in wt (JW2748), art1∆ (JW5232), 3nmt1-art1 (JW4889), and Art1-mECitrine (JW2694). (A) Arrows mark septa and arrowheads mark the contractile ring. (B) The numbers of Art1 and Rgf3 molecules globally in the whole cell and locally in mature but full-sized contractile rings. (C) Art1 affects Rgf3 protein levels. Anti-Myc antibody was used to detect Rgf3-13Myc (asterisks) in cell extracts from rgf3-13Myc (wt, JW2421) and rgf3-13Myc art1∆ (JW4816) strains grown in YE5S (top), or from rgf3-13Myc (wt, JW2421) and rgf3-13Myc 3nmt1-art1 (JW5129) strains induced in EMM5S for 24 h (bottom). The relative ratios of Rgf3 protein levels are given (n = 2). Scale bar: 5 μm.
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Related In: Results  -  Collection

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Figure 4: Art1 affects Rgf3 localization and protein levels. (A and B) Micrographs (A) and quantifications (B) of Rgf3-mECitrine in wt (JW2748), art1∆ (JW5232), 3nmt1-art1 (JW4889), and Art1-mECitrine (JW2694). (A) Arrows mark septa and arrowheads mark the contractile ring. (B) The numbers of Art1 and Rgf3 molecules globally in the whole cell and locally in mature but full-sized contractile rings. (C) Art1 affects Rgf3 protein levels. Anti-Myc antibody was used to detect Rgf3-13Myc (asterisks) in cell extracts from rgf3-13Myc (wt, JW2421) and rgf3-13Myc art1∆ (JW4816) strains grown in YE5S (top), or from rgf3-13Myc (wt, JW2421) and rgf3-13Myc 3nmt1-art1 (JW5129) strains induced in EMM5S for 24 h (bottom). The relative ratios of Rgf3 protein levels are given (n = 2). Scale bar: 5 μm.
Mentions: Because Art1 and Rgf3 interact with each other and Rgf3 plays a role in Art1 localization, it is interesting to know whether Art1 is involved in Rgf3 localization. In art1∆ cells, Rgf3 localization to the contractile ring was essentially abolished, and the intensity of Rgf3 in the cytoplasm was also reduced compared with wt (Figure 4A). However, a weak Rgf3 signal at the division site was detected in cells with a complete septum (Figure 4A, arrows). In contrast, Rgf3 intensities in the cytoplasm and the division site obviously increased when Art1 was overexpressed (Figure 4A, right).

Bottom Line: Using an Rgf3 conditional mutant and mislocalization experiments, we found that Art1 and Rgf3 are interdependent for localization to the division site.As expected, active Rho1 levels at the division site are reduced in art1∆ and rgf3 mutant cells.Taken together, these data reveal that the arrestin family protein Art1 regulates the protein levels and localization of the Rho-GEF Rgf3, which in turn modulates active Rho1 levels during fission yeast cytokinesis.

View Article: PubMed Central - PubMed

Affiliation: Graduate Program of Molecular, Cellular, and Developmental Biology, The Ohio State University, Columbus, OH 43210 Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210.

Show MeSH
Related in: MedlinePlus