Abstract
Expression of green fluorescent protein (GFP) linked to an actin binding domain is a commonly used method for live cell imaging of the actin cytoskeleton. One of these chimeric proteins is GFP-mTalin (GFP fused to the actin binding domain of mouse talin). Although it has been demonstrated that GFP-mTalin colocalizes with the actin cytoskeleton, its effect on actin dynamics and cell expansion has not been studied in detail. We created Arabidopsis (Arabidopsis thaliana) plants harboring alcohol inducible GFP-mTalin constructs to assess the effect of GFP-mTalin expression in vivo. We focused on the growing root hair as this is a model cell for studying cell expansion and root hair tip growth that requires a highly dynamic and polar actin cytoskeleton. We show that alcohol inducible expression of GFP-mTalin in root hairs causes severe defects in actin organization, resulting in either the termination of growth, cell death, and/or changes in cell shape. Fluorescence recovery after photobleaching experiments demonstrate that the interaction of GFP-mTalin and actin filaments is highly dynamic. To assess how GFP-mTalin affects actin dynamics we performed cosedimentation assays of GFP-mTalin with actin on its own or in the presence of the actin modulating protein, actin depolymerizing factor. We show that that GFP-mTalin does not affect actin polymerization but that it does inhibit the actin depolymerizing activity of actin depolymerizing factor. These observations demonstrate that GFP-mTalin can affect cell expansion, actin organization, and the interaction of actin binding proteins with actin.
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Claims 7
Claim · model-written summary
Induced GFP-mTalin expression caused severe root hair actin organization defects.
- Taxon
- Arabidopsis
Arabidopsist0 - Organ / stage
- root hairse0
- Treatment
- alcohol inducible expression of GFP-mTaline0
- Gene
- GFP-mTaline0
- Outcome
- actin organizatione0
- Direction
- severe defectse0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“alcohol inducible expression of GFP-mTalin in root hairs causes severe defects in actin organization”
t0“Arabidopsis”
Claim · model-written summary
Induced GFP-mTalin expression terminated root hair growth.
- Taxon
- ArabidopsisArabidopsist0
- Organ / stage
- root hairse0
- Treatment
- alcohol inducible expression of GFP-mTaline0
- Gene
- GFP-mTaline0
- Outcome
- termination of growthe0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“alcohol inducible expression of GFP-mTalin in root hairs causes severe defects in actin organization, resulting in either the termination of growth, cell death, and/or changes in cell shape.”
t0“Arabidopsis”
Claim · model-written summary
Induced GFP-mTalin expression caused root hair cell death.
- Taxon
- ArabidopsisArabidopsist0
- Organ / stage
- root hairse0
- Treatment
- alcohol inducible expression of GFP-mTaline0
- Gene
- GFP-mTaline0
- Outcome
- cell deathe0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“alcohol inducible expression of GFP-mTalin in root hairs causes severe defects in actin organization, resulting in either the termination of growth, cell death, and/or changes in cell shape.”
t0“Arabidopsis”
Claim · model-written summary
Induced GFP-mTalin expression changed root hair cell shape.
- Taxon
- ArabidopsisArabidopsist0
- Organ / stage
- root hairse0
- Treatment
- alcohol inducible expression of GFP-mTaline0
- Gene
- GFP-mTaline0
- Outcome
- cell shapee0
- Direction
- changese0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“alcohol inducible expression of GFP-mTalin in root hairs causes severe defects in actin organization, resulting in either the termination of growth, cell death, and/or changes in cell shape.”
t0“Arabidopsis”
Claim · model-written summary
GFP-mTalin interacted dynamically with actin filaments.
- Taxon
- ArabidopsisArabidopsist0
- Gene
- GFP-mTaline0
- Outcome
- interaction of GFP-mTalin and actin filamentse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the interaction of GFP-mTalin and actin filaments is highly dynamic.”
t0“Arabidopsis”
Claim · model-written summary
GFP-mTalin did not affect actin polymerization in the assay.
- Taxon
- ArabidopsisArabidopsist0
- Treatment
- GFP-mTaline0
- Outcome
- actin polymerizatione0
- Direction
- does not affecte0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GFP-mTalin does not affect actin polymerization”
t0“Arabidopsis”
Claim · model-written summary
GFP-mTalin inhibited actin depolymerizing factor activity in the assay.
- Taxon
- ArabidopsisArabidopsist0
- Treatment
- GFP-mTaline1
- Gene
- actin depolymerizing factore0
- Outcome
- actin depolymerizing activity of actin depolymerizing factore0
- Direction
- inhibite0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“it does inhibit the actin depolymerizing activity of actin depolymerizing factor.”
e1“GFP-mTalin does not affect actin polymerization”
t0“Arabidopsis”
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