Source 1primary paper2013Cytoskeleton
Toolkit/photoactivatable diaphanous autoregulatory domain
photoactivatable diaphanous autoregulatory domain
Also known as: caged diaphanous auto-regulatory domain, optogenetic technique for the activation of diaphanous-related formins
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The photoactivatable diaphanous autoregulatory domain is an optogenetic fusion of the Avena sativa Phototropin1 LOV2 domain to an isolated mDia1 diaphanous autoregulatory domain. In blue light, this caged construct rapidly activates endogenous diaphanous-related formins, whereas it is inactive in the dark.
Usefulness & Problems
Why this is useful
This tool enables acute optical control of endogenous diaphanous-related formin activity using blue light. In the reported study, photoactivation induced filopodia and lamellipodia formation and increased F-actin along existing stress fibers within minutes, supporting perturbation of actin cytoskeletal dynamics with temporal precision.
Source:
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
Source:
demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous-related formin function in cells
Problem solved
It addresses the problem of rapidly and reversibly activating endogenous diaphanous-related formins without constitutive activity in the dark. The reported application was to probe how formin-driven F-actin assembly affects stress fiber architecture and contractility.
Source:
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
activation of endogenous diaphanous-related forminsallosteric switchinglight-gated uncagingTechniques
No technique tags yet.
Target processes
recombinationInput: Light
Implementation Constraints
The tool is implemented as a fusion between the Avena sativa Phototropin1 LOV2 domain and an isolated mDia1 diaphanous autoregulatory domain. Its active input is blue light, and the reported mode of action is activation of endogenous diaphanous-related formins rather than delivery of an exogenous catalytic effector. The supplied evidence does not specify construct architecture details beyond the fusion partners, nor expression system or delivery method.
The evidence provided comes from a single 2013 Cytoskeleton study, so validation breadth is limited. The supplied evidence does not report quantitative activation kinetics, reversibility, spectral tuning beyond blue light, or performance across multiple cell types or organisms.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Photo-activation of the tool induced filopodia and lamellipodia formation and increased F-actin along existing stress fibers within minutes.
Using an F-actin reporter, we observed filopodia and lamellipodia formation as well as a steady increase in F-actin along existing stress fibers, starting within minutes of photo-activation.
response onset time within minutes
Photo-activation of the tool induced filopodia and lamellipodia formation and increased F-actin along existing stress fibers within minutes.
Using an F-actin reporter, we observed filopodia and lamellipodia formation as well as a steady increase in F-actin along existing stress fibers, starting within minutes of photo-activation.
response onset time within minutes
Photo-activation of the tool induced filopodia and lamellipodia formation and increased F-actin along existing stress fibers within minutes.
Using an F-actin reporter, we observed filopodia and lamellipodia formation as well as a steady increase in F-actin along existing stress fibers, starting within minutes of photo-activation.
response onset time within minutes
Photo-activation of the tool induced filopodia and lamellipodia formation and increased F-actin along existing stress fibers within minutes.
Using an F-actin reporter, we observed filopodia and lamellipodia formation as well as a steady increase in F-actin along existing stress fibers, starting within minutes of photo-activation.
response onset time within minutes
Photo-activation of the tool induced filopodia and lamellipodia formation and increased F-actin along existing stress fibers within minutes.
Using an F-actin reporter, we observed filopodia and lamellipodia formation as well as a steady increase in F-actin along existing stress fibers, starting within minutes of photo-activation.
response onset time within minutes
Photo-activation of the tool induced filopodia and lamellipodia formation and increased F-actin along existing stress fibers within minutes.
Using an F-actin reporter, we observed filopodia and lamellipodia formation as well as a steady increase in F-actin along existing stress fibers, starting within minutes of photo-activation.
response onset time within minutes
Photo-activation of the tool induced filopodia and lamellipodia formation and increased F-actin along existing stress fibers within minutes.
Using an F-actin reporter, we observed filopodia and lamellipodia formation as well as a steady increase in F-actin along existing stress fibers, starting within minutes of photo-activation.
response onset time within minutes
The caged diaphanous autoregulatory domain was inactive in the dark and rapidly activated endogenous diaphanous-related formins in blue light.
This "caged" diaphanous auto-regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous-related formins.
The caged diaphanous autoregulatory domain was inactive in the dark and rapidly activated endogenous diaphanous-related formins in blue light.
This "caged" diaphanous auto-regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous-related formins.
The caged diaphanous autoregulatory domain was inactive in the dark and rapidly activated endogenous diaphanous-related formins in blue light.
This "caged" diaphanous auto-regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous-related formins.
The caged diaphanous autoregulatory domain was inactive in the dark and rapidly activated endogenous diaphanous-related formins in blue light.
This "caged" diaphanous auto-regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous-related formins.
The caged diaphanous autoregulatory domain was inactive in the dark and rapidly activated endogenous diaphanous-related formins in blue light.
This "caged" diaphanous auto-regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous-related formins.
The caged diaphanous autoregulatory domain was inactive in the dark and rapidly activated endogenous diaphanous-related formins in blue light.
This "caged" diaphanous auto-regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous-related formins.
The caged diaphanous autoregulatory domain was inactive in the dark and rapidly activated endogenous diaphanous-related formins in blue light.
This "caged" diaphanous auto-regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous-related formins.
The results suggest a decoupling between F-actin accumulation and contractility in stress fibers.
Our results suggest a decoupling between F-actin accumulation and contractility in stress fibers
The results suggest a decoupling between F-actin accumulation and contractility in stress fibers.
Our results suggest a decoupling between F-actin accumulation and contractility in stress fibers
The results suggest a decoupling between F-actin accumulation and contractility in stress fibers.
Our results suggest a decoupling between F-actin accumulation and contractility in stress fibers
The results suggest a decoupling between F-actin accumulation and contractility in stress fibers.
Our results suggest a decoupling between F-actin accumulation and contractility in stress fibers
The results suggest a decoupling between F-actin accumulation and contractility in stress fibers.
Our results suggest a decoupling between F-actin accumulation and contractility in stress fibers
The results suggest a decoupling between F-actin accumulation and contractility in stress fibers.
Our results suggest a decoupling between F-actin accumulation and contractility in stress fibers
The results suggest a decoupling between F-actin accumulation and contractility in stress fibers.
Our results suggest a decoupling between F-actin accumulation and contractility in stress fibers
Photo-activation did not induce formation of new stress fibers.
Interestingly, we did not observe the formation of new stress fibers.
Photo-activation did not induce formation of new stress fibers.
Interestingly, we did not observe the formation of new stress fibers.
Photo-activation did not induce formation of new stress fibers.
Interestingly, we did not observe the formation of new stress fibers.
Photo-activation did not induce formation of new stress fibers.
Interestingly, we did not observe the formation of new stress fibers.
Photo-activation did not induce formation of new stress fibers.
Interestingly, we did not observe the formation of new stress fibers.
Photo-activation did not induce formation of new stress fibers.
Interestingly, we did not observe the formation of new stress fibers.
Photo-activation did not induce formation of new stress fibers.
Interestingly, we did not observe the formation of new stress fibers.
A 1.9-fold increase in F-actin along stress fibers was not accompanied by increased myosin II or an apparent increase in tension judged by focal adhesion size.
Remarkably, a 1.9-fold increase in F-actin was not paralleled by an increase in myosin II along stress fibers and the amount of tension generated by the fibers, as judged by focal adhesion size, appeared unchanged.
F-actin increase 1.9 fold
A 1.9-fold increase in F-actin along stress fibers was not accompanied by increased myosin II or an apparent increase in tension judged by focal adhesion size.
Remarkably, a 1.9-fold increase in F-actin was not paralleled by an increase in myosin II along stress fibers and the amount of tension generated by the fibers, as judged by focal adhesion size, appeared unchanged.
F-actin increase 1.9 fold
A 1.9-fold increase in F-actin along stress fibers was not accompanied by increased myosin II or an apparent increase in tension judged by focal adhesion size.
Remarkably, a 1.9-fold increase in F-actin was not paralleled by an increase in myosin II along stress fibers and the amount of tension generated by the fibers, as judged by focal adhesion size, appeared unchanged.
F-actin increase 1.9 fold
A 1.9-fold increase in F-actin along stress fibers was not accompanied by increased myosin II or an apparent increase in tension judged by focal adhesion size.
Remarkably, a 1.9-fold increase in F-actin was not paralleled by an increase in myosin II along stress fibers and the amount of tension generated by the fibers, as judged by focal adhesion size, appeared unchanged.
F-actin increase 1.9 fold
A 1.9-fold increase in F-actin along stress fibers was not accompanied by increased myosin II or an apparent increase in tension judged by focal adhesion size.
Remarkably, a 1.9-fold increase in F-actin was not paralleled by an increase in myosin II along stress fibers and the amount of tension generated by the fibers, as judged by focal adhesion size, appeared unchanged.
F-actin increase 1.9 fold
A 1.9-fold increase in F-actin along stress fibers was not accompanied by increased myosin II or an apparent increase in tension judged by focal adhesion size.
Remarkably, a 1.9-fold increase in F-actin was not paralleled by an increase in myosin II along stress fibers and the amount of tension generated by the fibers, as judged by focal adhesion size, appeared unchanged.
F-actin increase 1.9 fold
A 1.9-fold increase in F-actin along stress fibers was not accompanied by increased myosin II or an apparent increase in tension judged by focal adhesion size.
Remarkably, a 1.9-fold increase in F-actin was not paralleled by an increase in myosin II along stress fibers and the amount of tension generated by the fibers, as judged by focal adhesion size, appeared unchanged.
F-actin increase 1.9 fold
The authors developed an optogenetic technique for activation of endogenous diaphanous-related formins based on a LOV2-mDia1 diaphanous autoregulatory domain fusion.
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
The authors developed an optogenetic technique for activation of endogenous diaphanous-related formins based on a LOV2-mDia1 diaphanous autoregulatory domain fusion.
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
The authors developed an optogenetic technique for activation of endogenous diaphanous-related formins based on a LOV2-mDia1 diaphanous autoregulatory domain fusion.
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
The authors developed an optogenetic technique for activation of endogenous diaphanous-related formins based on a LOV2-mDia1 diaphanous autoregulatory domain fusion.
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
The authors developed an optogenetic technique for activation of endogenous diaphanous-related formins based on a LOV2-mDia1 diaphanous autoregulatory domain fusion.
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
The authors developed an optogenetic technique for activation of endogenous diaphanous-related formins based on a LOV2-mDia1 diaphanous autoregulatory domain fusion.
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
The authors developed an optogenetic technique for activation of endogenous diaphanous-related formins based on a LOV2-mDia1 diaphanous autoregulatory domain fusion.
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
The photoactivatable diaphanous autoregulatory domain is useful for studying diaphanous-related formin function in cells.
demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous-related formin function in cells
The photoactivatable diaphanous autoregulatory domain is useful for studying diaphanous-related formin function in cells.
demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous-related formin function in cells
The photoactivatable diaphanous autoregulatory domain is useful for studying diaphanous-related formin function in cells.
demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous-related formin function in cells
The photoactivatable diaphanous autoregulatory domain is useful for studying diaphanous-related formin function in cells.
demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous-related formin function in cells
The photoactivatable diaphanous autoregulatory domain is useful for studying diaphanous-related formin function in cells.
demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous-related formin function in cells
The photoactivatable diaphanous autoregulatory domain is useful for studying diaphanous-related formin function in cells.
demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous-related formin function in cells
The photoactivatable diaphanous autoregulatory domain is useful for studying diaphanous-related formin function in cells.
demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous-related formin function in cells
Approval Evidence
1 source7 linked approval claimsfirst-pass slug photoactivatable-diaphanous-autoregulatory-domain
Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1. This "caged" diaphanous auto-regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous-related formins.
Source:
cellular effectsupports
Photo-activation of the tool induced filopodia and lamellipodia formation and increased F-actin along existing stress fibers within minutes.
Using an F-actin reporter, we observed filopodia and lamellipodia formation as well as a steady increase in F-actin along existing stress fibers, starting within minutes of photo-activation.
Source:
light responsivenesssupports
The caged diaphanous autoregulatory domain was inactive in the dark and rapidly activated endogenous diaphanous-related formins in blue light.
This "caged" diaphanous auto-regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous-related formins.
Source:
mechanistic interpretationsupports
The results suggest a decoupling between F-actin accumulation and contractility in stress fibers.
Our results suggest a decoupling between F-actin accumulation and contractility in stress fibers
Source:
negative observationsupports
Photo-activation did not induce formation of new stress fibers.
Interestingly, we did not observe the formation of new stress fibers.
Source:
quantitative effectsupports
A 1.9-fold increase in F-actin along stress fibers was not accompanied by increased myosin II or an apparent increase in tension judged by focal adhesion size.
Remarkably, a 1.9-fold increase in F-actin was not paralleled by an increase in myosin II along stress fibers and the amount of tension generated by the fibers, as judged by focal adhesion size, appeared unchanged.
Source:
tool developmentsupports
The authors developed an optogenetic technique for activation of endogenous diaphanous-related formins based on a LOV2-mDia1 diaphanous autoregulatory domain fusion.
We have developed an optogenetic technique for the activation of diaphanous-related formins. Our approach is based on fusion of the light-oxygen-voltage 2 domain of Avena sativa Phototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1.
Source:
utility claimsupports
The photoactivatable diaphanous autoregulatory domain is useful for studying diaphanous-related formin function in cells.
demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous-related formin function in cells
Source:
Comparisons
Source-backed strengths
The construct was reported to be inactive in the dark and rapidly active under blue light, indicating strong light dependence and fast switching. Cellular validation showed induction of filopodia, lamellipodia, and increased F-actin along existing stress fibers within minutes. The associated study further suggested that stress fiber thickening could occur without an increase in contractility.
Ranked Citations
- 1.