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Genetic suppression of a phosphomimic myosin II identifies system-level factors that promote myosin II cleavage furrow accumulation.

Ren Y, West-Foyle H, Surcel A, Miller C, Robinson DN - Mol. Biol. Cell (2014)

Bottom Line: How myosin II localizes to the cleavage furrow in Dictyostelium and metazoan cells remains largely unknown despite significant advances in understanding its regulation.Finally, an engineered myosin II with a longer lever arm (2xELC), producing a highly mechanosensitive motor, could also partially suppress the intragenic 3xAsp.Overall, myosin II accumulation is the result of multiple parallel and partially redundant pathways that comprise a cellular contractility control system.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.

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Depletion of rmd1 mRNA reduced GFP-myosin II cleavage furrow accumulation. (A) Micrographs show GFP-myosin II localization at the cleavage furrow cortex of WT control (myoII::GFP-myosin II; pLD1) and rmd1hp (myoII::GFP-myosin II; rmd1hp) cells. Scale bar, 10 μm. (B) Bar graph shows that the GFP-myosin II intensity at the cleavage furrow cortex is reduced in the rmd1hp cells compared with control (cell lines are the same as in A). Sample sizes are displayed in the bar graphs. Measurements are derived from 36 total control cells and 47 total rmd1hp cells. The differences between control and rmd1hp are significant (ANOVA, p < 0.0001). (C) Micrographs show myosin II accumulation at the cleavage furrow of cells compressed by agarose overlay. Scale bar, 10 μm. (D) WT control and rmd1hp cells showed comparable cleavage furrow accumulation when mechanical stress was applied. Because the data were similar across all stages of cytokinesis, the data were combined into a single group for each genotype. The inset shows the relative rmd1 mRNA levels in WT control and rmd1hp cells, which were used for the analyses in A–D.
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Figure 6: Depletion of rmd1 mRNA reduced GFP-myosin II cleavage furrow accumulation. (A) Micrographs show GFP-myosin II localization at the cleavage furrow cortex of WT control (myoII::GFP-myosin II; pLD1) and rmd1hp (myoII::GFP-myosin II; rmd1hp) cells. Scale bar, 10 μm. (B) Bar graph shows that the GFP-myosin II intensity at the cleavage furrow cortex is reduced in the rmd1hp cells compared with control (cell lines are the same as in A). Sample sizes are displayed in the bar graphs. Measurements are derived from 36 total control cells and 47 total rmd1hp cells. The differences between control and rmd1hp are significant (ANOVA, p < 0.0001). (C) Micrographs show myosin II accumulation at the cleavage furrow of cells compressed by agarose overlay. Scale bar, 10 μm. (D) WT control and rmd1hp cells showed comparable cleavage furrow accumulation when mechanical stress was applied. Because the data were similar across all stages of cytokinesis, the data were combined into a single group for each genotype. The inset shows the relative rmd1 mRNA levels in WT control and rmd1hp cells, which were used for the analyses in A–D.

Mentions: bData for late-stage furrows from Figure 6B.


Genetic suppression of a phosphomimic myosin II identifies system-level factors that promote myosin II cleavage furrow accumulation.

Ren Y, West-Foyle H, Surcel A, Miller C, Robinson DN - Mol. Biol. Cell (2014)

Depletion of rmd1 mRNA reduced GFP-myosin II cleavage furrow accumulation. (A) Micrographs show GFP-myosin II localization at the cleavage furrow cortex of WT control (myoII::GFP-myosin II; pLD1) and rmd1hp (myoII::GFP-myosin II; rmd1hp) cells. Scale bar, 10 μm. (B) Bar graph shows that the GFP-myosin II intensity at the cleavage furrow cortex is reduced in the rmd1hp cells compared with control (cell lines are the same as in A). Sample sizes are displayed in the bar graphs. Measurements are derived from 36 total control cells and 47 total rmd1hp cells. The differences between control and rmd1hp are significant (ANOVA, p < 0.0001). (C) Micrographs show myosin II accumulation at the cleavage furrow of cells compressed by agarose overlay. Scale bar, 10 μm. (D) WT control and rmd1hp cells showed comparable cleavage furrow accumulation when mechanical stress was applied. Because the data were similar across all stages of cytokinesis, the data were combined into a single group for each genotype. The inset shows the relative rmd1 mRNA levels in WT control and rmd1hp cells, which were used for the analyses in A–D.
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Related In: Results  -  Collection

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Figure 6: Depletion of rmd1 mRNA reduced GFP-myosin II cleavage furrow accumulation. (A) Micrographs show GFP-myosin II localization at the cleavage furrow cortex of WT control (myoII::GFP-myosin II; pLD1) and rmd1hp (myoII::GFP-myosin II; rmd1hp) cells. Scale bar, 10 μm. (B) Bar graph shows that the GFP-myosin II intensity at the cleavage furrow cortex is reduced in the rmd1hp cells compared with control (cell lines are the same as in A). Sample sizes are displayed in the bar graphs. Measurements are derived from 36 total control cells and 47 total rmd1hp cells. The differences between control and rmd1hp are significant (ANOVA, p < 0.0001). (C) Micrographs show myosin II accumulation at the cleavage furrow of cells compressed by agarose overlay. Scale bar, 10 μm. (D) WT control and rmd1hp cells showed comparable cleavage furrow accumulation when mechanical stress was applied. Because the data were similar across all stages of cytokinesis, the data were combined into a single group for each genotype. The inset shows the relative rmd1 mRNA levels in WT control and rmd1hp cells, which were used for the analyses in A–D.
Mentions: bData for late-stage furrows from Figure 6B.

Bottom Line: How myosin II localizes to the cleavage furrow in Dictyostelium and metazoan cells remains largely unknown despite significant advances in understanding its regulation.Finally, an engineered myosin II with a longer lever arm (2xELC), producing a highly mechanosensitive motor, could also partially suppress the intragenic 3xAsp.Overall, myosin II accumulation is the result of multiple parallel and partially redundant pathways that comprise a cellular contractility control system.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.

Show MeSH
Related in: MedlinePlus