Source 1primary paper2020PLoS Biology
Toolkit/engineered GEF-Pak1 interaction
engineered GEF-Pak1 interaction
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The engineered GEF-Pak1 interaction is a rewired Cdc42 positive-feedback construct in Schizosaccharomyces pombe in which a guanine nucleotide exchange factor is engineered to interact with the Cdc42 effector p21-activated kinase 1 (Pak1). This engineered coupling supports scaffold-mediated positive feedback, promotes active Cdc42 zone formation, and enables rod-shape polarization.
Usefulness & Problems
Why this is useful
This construct is useful for dissecting how scaffold-mediated positive feedback contributes to cell polarization downstream of Cdc42. The reported system also provides a way to test Ras-gated robustness of polarization and to probe polarity network sufficiency when native activator dependencies are altered.
Problem solved
It addresses the problem of determining whether Cdc42 positive feedback can be rewired through an engineered GEF-effector interaction to support polarization. The cited work indicates that this rewiring can sustain viability and active Cdc42 zone formation even when otherwise essential Cdc42 activators are absent.
Problem links
Need conditional control of signaling activity
DerivedThe engineered GEF-Pak1 interaction is a rewired Cdc42 positive-feedback construct in Schizosaccharomyces pombe in which a guanine nucleotide exchange factor is engineered to interact with the Cdc42 effector p21-activated kinase 1 (Pak1). This engineered coupling supports scaffold-mediated positive feedback, promotes active Cdc42 zone formation, and enables rod-shape polarization.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
engineered gef-effector couplinglight-dependent protein recruitmentras-gated polarizationscaffold-mediated positive feedbackTechniques
No technique tags yet.
Target processes
signalingImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: regulatorswitch architecture: recruitment
The construct was studied in Schizosaccharomyces pombe in the context of Cdc42 polarity control and Pak1 effector signaling. The source literature involved optogenetics and light-dependent protein recruitment, but the supplied evidence does not define wavelengths, domains, cofactors, or detailed construct design parameters for the engineered GEF-Pak1 interaction itself.
The evidence provided comes from a single 2020 study in Schizosaccharomyces pombe and does not establish performance in other organisms or signaling contexts. The supplied evidence does not specify the exact GEF identity, molecular architecture of the engineered interface, quantitative kinetics, or implementation constraints beyond the rewired interaction concept.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successYeastapplication demoSchizosaccharomyces pombe
Inferred from claim c4 during normalization. The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking. Derived from claim c4. Quoted text: this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
Source:
successYeastapplication demoSchizosaccharomyces pombe
Inferred from claim c4 during normalization. The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking. Derived from claim c4. Quoted text: this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
Source:
successYeastapplication demoSchizosaccharomyces pombe
Inferred from claim c4 during normalization. The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking. Derived from claim c4. Quoted text: this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
Source:
successYeastapplication demoSchizosaccharomyces pombe
Inferred from claim c4 during normalization. The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking. Derived from claim c4. Quoted text: this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
Source:
successYeastapplication demoSchizosaccharomyces pombe
Inferred from claim c4 during normalization. The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking. Derived from claim c4. Quoted text: this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
Source:
successYeastapplication demoSchizosaccharomyces pombe
Inferred from claim c4 during normalization. The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking. Derived from claim c4. Quoted text: this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
Source:
successYeastapplication demoSchizosaccharomyces pombe
Inferred from claim c4 during normalization. The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking. Derived from claim c4. Quoted text: this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
Source:
Supporting Sources
Ranked Claims
Scaffold-mediated positive feedback gated by Ras activity confers robust polarization for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, confers robust polarization for rod-shape formation.
Scaffold-mediated positive feedback gated by Ras activity confers robust polarization for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, confers robust polarization for rod-shape formation.
Scaffold-mediated positive feedback gated by Ras activity confers robust polarization for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, confers robust polarization for rod-shape formation.
Scaffold-mediated positive feedback gated by Ras activity confers robust polarization for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, confers robust polarization for rod-shape formation.
Scaffold-mediated positive feedback gated by Ras activity confers robust polarization for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, confers robust polarization for rod-shape formation.
Scaffold-mediated positive feedback gated by Ras activity confers robust polarization for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, confers robust polarization for rod-shape formation.
Scaffold-mediated positive feedback gated by Ras activity confers robust polarization for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, confers robust polarization for rod-shape formation.
Scd2 is dispensable when positive feedback is rewired through an engineered interaction between the GEF and Pak1.
Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
Scd2 is dispensable when positive feedback is rewired through an engineered interaction between the GEF and Pak1.
Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
Scd2 is dispensable when positive feedback is rewired through an engineered interaction between the GEF and Pak1.
Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
Scd2 is dispensable when positive feedback is rewired through an engineered interaction between the GEF and Pak1.
Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
Scd2 is dispensable when positive feedback is rewired through an engineered interaction between the GEF and Pak1.
Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
Scd2 is dispensable when positive feedback is rewired through an engineered interaction between the GEF and Pak1.
Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
Scd2 is dispensable when positive feedback is rewired through an engineered interaction between the GEF and Pak1.
Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking.
this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking.
this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking.
this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking.
this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking.
this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking.
this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking.
this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
The study implemented the CRY2-CIB1 optogenetic system for acute light-dependent protein recruitment to the plasma membrane in Schizosaccharomyces pombe.
We implemented the CRY2-CIB1 optogenetic system for acute light-dependent protein recruitment to the plasma membrane
Optogenetic recruitment of constitutively active Cdc42 leads to co-recruitment of Scd1 and endogenous Cdc42 in a Scd2-dependent manner.
optogenetic recruitment of constitutively active Cdc42 leads to co-recruitment of the guanine nucleotide exchange factor (GEF) Scd1 and endogenous Cdc42, in a manner dependent on the scaffold protein Scd2
Ras1 localizes Scd1 and promotes its activity, thereby potentiating positive feedback.
the small GTPase Ras1 plays a role in both localizing the GEF Scd1 and promoting its activity, which potentiates the positive feedback
Ras1 localizes Scd1 and promotes its activity, thereby potentiating positive feedback.
the small GTPase Ras1 plays a role in both localizing the GEF Scd1 and promoting its activity, which potentiates the positive feedback
Ras1 localizes Scd1 and promotes its activity, thereby potentiating positive feedback.
the small GTPase Ras1 plays a role in both localizing the GEF Scd1 and promoting its activity, which potentiates the positive feedback
Ras1 localizes Scd1 and promotes its activity, thereby potentiating positive feedback.
the small GTPase Ras1 plays a role in both localizing the GEF Scd1 and promoting its activity, which potentiates the positive feedback
Ras1 localizes Scd1 and promotes its activity, thereby potentiating positive feedback.
the small GTPase Ras1 plays a role in both localizing the GEF Scd1 and promoting its activity, which potentiates the positive feedback
Ras1 localizes Scd1 and promotes its activity, thereby potentiating positive feedback.
the small GTPase Ras1 plays a role in both localizing the GEF Scd1 and promoting its activity, which potentiates the positive feedback
Ras1 localizes Scd1 and promotes its activity, thereby potentiating positive feedback.
the small GTPase Ras1 plays a role in both localizing the GEF Scd1 and promoting its activity, which potentiates the positive feedback
Approval Evidence
1 source4 linked approval claimsfirst-pass slug engineered-gef-pak1-interaction
the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
Source:
conclusionsupports
Scaffold-mediated positive feedback gated by Ras activity confers robust polarization for rod-shape formation.
We conclude that scaffold-mediated positive feedback, gated by Ras activity, confers robust polarization for rod-shape formation.
Source:
engineering resultsupports
Scd2 is dispensable when positive feedback is rewired through an engineered interaction between the GEF and Pak1.
Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
Source:
engineering resultsupports
The rewired positive feedback confers viability and allows formation of two zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking.
this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
Source:
mechanismsupports
Ras1 localizes Scd1 and promotes its activity, thereby potentiating positive feedback.
the small GTPase Ras1 plays a role in both localizing the GEF Scd1 and promoting its activity, which potentiates the positive feedback
Source:
Comparisons
Source-backed strengths
The engineered interaction was reported to confer viability and to allow formation of two zones of active Cdc42 in cells lacking otherwise essential Cdc42 activators. The same study concluded that scaffold-mediated positive feedback gated by Ras activity confers robust polarization for rod-shape formation, and that Scd2 becomes dispensable when feedback is rewired through the engineered GEF-Pak1 interaction.
Source:
Scd2 function is dispensable when the positive feedback operates through an engineered interaction between the GEF and a Cdc42 effector, the p21-activated kinase 1 (Pak1)
Source:
this rewired positive feedback confers viability and allows cells to form 2 zones of active Cdc42 even when otherwise essential Cdc42 activators are lacking
Compared with genetically encoded FRET biosensor
engineered GEF-Pak1 interaction and genetically encoded FRET biosensor address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling
Compared with kinase translocation reporters
engineered GEF-Pak1 interaction and kinase translocation reporters address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling
engineered GEF-Pak1 interaction and novel fluorescent biosensor for mitochondrial outer membrane rupture address a similar problem space because they share signaling.
Shared frame: same top-level item type; shared target processes: signaling
Ranked Citations
- 1.