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A direct interaction between fascin and microtubules contributes to adhesion dynamics and cell migration.

Villari G, Jayo A, Zanet J, Fitch B, Serrels B, Frame M, Stramer BM, Goult BT, Parsons M - J. Cell. Sci. (2015)

Bottom Line: However, potential non-actin bundling roles for fascin remain unknown.Here, we show for the first time that fascin can directly interact with the microtubule cytoskeleton and that this does not depend upon fascin-actin bundling.These findings shed light on new non actin-dependent roles for fascin and might have implications for the design of therapies to target fascin in metastatic disease.

View Article: PubMed Central - PubMed

Affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, Guys Campus, London SE1 1UL, UK.

No MeSH data available.


Related in: MedlinePlus

Fascin–MT binding regulates cell adhesion dynamics and migration. (A) Quantification of focal adhesion surface area coverage of MDA MB 231 FascinKD cells expressing WT or mutant GFP–fascin that were either untreated (UT), treated with NOC for 20 min (NOC) or at 60 min post-NOC washout (60′W). n=45 cells were quantified across three independent experiments. (B) Example migration tracks of MDA MB 231 FascinKD cells expressing WT or mutant GFP–fascin taken from time-lapse movies. (C) Quantification of migration speed as determined from tracks as shown in C. n=60 cells per condition. Mean±s.e.m. values are shown. *P<0.05; **P<0.01 compared to controls (one-way ANOVA).
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JCS175760F4: Fascin–MT binding regulates cell adhesion dynamics and migration. (A) Quantification of focal adhesion surface area coverage of MDA MB 231 FascinKD cells expressing WT or mutant GFP–fascin that were either untreated (UT), treated with NOC for 20 min (NOC) or at 60 min post-NOC washout (60′W). n=45 cells were quantified across three independent experiments. (B) Example migration tracks of MDA MB 231 FascinKD cells expressing WT or mutant GFP–fascin taken from time-lapse movies. (C) Quantification of migration speed as determined from tracks as shown in C. n=60 cells per condition. Mean±s.e.m. values are shown. *P<0.05; **P<0.01 compared to controls (one-way ANOVA).

Mentions: To determine whether these MT-binding differences resulted in adhesion area changes, we quantified adhesion area in cells expressing S39A, S39D, S274A or S274D mutants of fascin. Data demonstrated that S39A, S39D and S274A mutants of fascin were able to support adhesion disassembly in response to NOC washout (Fig. 4A; Fig. S3B). Conversely, cells expressing S274D-fascin showed larger adhesions under both basal conditions and upon NOC washout (Fig. 4A), mimicking the effect seen in cells expressing the MT1-fascin mutant, which also similarly exhibits increased MT binding (Fig. 2). There were no changes in total adhesion protein levels in cells expressing fascin mutants (Fig. S3C). In agreement with the MT re-growth assay data, the adhesion disassembly defect in S274D cells was partially reversed in the S39A/S274D double mutant cells, but not in the S39D/S274D cells (Fig. 4A). These data together further suggest that S274 plays an important role in fascin–MT association and focal adhesion dynamics, potentially through disconnecting fascin from F-actin and promoting direct fascin­–MT binding.Fig. 4.


A direct interaction between fascin and microtubules contributes to adhesion dynamics and cell migration.

Villari G, Jayo A, Zanet J, Fitch B, Serrels B, Frame M, Stramer BM, Goult BT, Parsons M - J. Cell. Sci. (2015)

Fascin–MT binding regulates cell adhesion dynamics and migration. (A) Quantification of focal adhesion surface area coverage of MDA MB 231 FascinKD cells expressing WT or mutant GFP–fascin that were either untreated (UT), treated with NOC for 20 min (NOC) or at 60 min post-NOC washout (60′W). n=45 cells were quantified across three independent experiments. (B) Example migration tracks of MDA MB 231 FascinKD cells expressing WT or mutant GFP–fascin taken from time-lapse movies. (C) Quantification of migration speed as determined from tracks as shown in C. n=60 cells per condition. Mean±s.e.m. values are shown. *P<0.05; **P<0.01 compared to controls (one-way ANOVA).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4696496&req=5

JCS175760F4: Fascin–MT binding regulates cell adhesion dynamics and migration. (A) Quantification of focal adhesion surface area coverage of MDA MB 231 FascinKD cells expressing WT or mutant GFP–fascin that were either untreated (UT), treated with NOC for 20 min (NOC) or at 60 min post-NOC washout (60′W). n=45 cells were quantified across three independent experiments. (B) Example migration tracks of MDA MB 231 FascinKD cells expressing WT or mutant GFP–fascin taken from time-lapse movies. (C) Quantification of migration speed as determined from tracks as shown in C. n=60 cells per condition. Mean±s.e.m. values are shown. *P<0.05; **P<0.01 compared to controls (one-way ANOVA).
Mentions: To determine whether these MT-binding differences resulted in adhesion area changes, we quantified adhesion area in cells expressing S39A, S39D, S274A or S274D mutants of fascin. Data demonstrated that S39A, S39D and S274A mutants of fascin were able to support adhesion disassembly in response to NOC washout (Fig. 4A; Fig. S3B). Conversely, cells expressing S274D-fascin showed larger adhesions under both basal conditions and upon NOC washout (Fig. 4A), mimicking the effect seen in cells expressing the MT1-fascin mutant, which also similarly exhibits increased MT binding (Fig. 2). There were no changes in total adhesion protein levels in cells expressing fascin mutants (Fig. S3C). In agreement with the MT re-growth assay data, the adhesion disassembly defect in S274D cells was partially reversed in the S39A/S274D double mutant cells, but not in the S39D/S274D cells (Fig. 4A). These data together further suggest that S274 plays an important role in fascin–MT association and focal adhesion dynamics, potentially through disconnecting fascin from F-actin and promoting direct fascin­–MT binding.Fig. 4.

Bottom Line: However, potential non-actin bundling roles for fascin remain unknown.Here, we show for the first time that fascin can directly interact with the microtubule cytoskeleton and that this does not depend upon fascin-actin bundling.These findings shed light on new non actin-dependent roles for fascin and might have implications for the design of therapies to target fascin in metastatic disease.

View Article: PubMed Central - PubMed

Affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, Guys Campus, London SE1 1UL, UK.

No MeSH data available.


Related in: MedlinePlus