Source 1primary paper2022MED
Toolkit/BcWCL1 PASΔ
BcWCL1 PASΔ
Also known as: BcWCL1 PAS deletion, BcWCL1<PASΔ>, BcWCL1PASΔ
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
BcWCL1 PASΔ is a PAS-domain-deleted variant of the Botrytis cinerea blue-light photoreceptor BcWCL1 that functions in yeast as a blue-light-activated transcription switch. The reported activity depends on the BcWCL1 N-terminal region, which supports light-stimulated self-dimerization and contains a functional 9aaTAD-family transcriptional activation domain.
Usefulness & Problems
Why this is useful
This tool is useful as a light-responsive transcriptional regulator that couples blue-light sensing to gene activation in yeast. Its activation domain was reported to produce transcriptional activation levels comparable to Gal4 and p65 activation domains, supporting its use where strong inducible transcription is desired.
Source:
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
Problem solved
BcWCL1 PASΔ addresses the need for an optogenetic transcription switch that can be activated by blue light and encoded within a BcWCL1-derived architecture. The available evidence specifically supports its use for controlling transcription in yeast through light-stimulated dimerization and activation-domain function.
Published Workflows
Objective: Map the regions of BcWCL1 PASΔ responsible for blue-light-dependent self-dimerization and transcriptional activation in yeast and assess its optogenetic utility.
Why it works: The workflow tests different N-terminal and C-terminal fragments in a reporter-based yeast system so that light-dependent dimerization and transcriptional activation can be attributed to specific regions of BcWCL1 PASΔ.
light-stimulated self-dimerization through the N-terminal regionN-terminal transcriptional activation domain functionchimeric transcription factorsluciferase reporter assayfragment-based domain mapping
Stages
- 1.Fragment generation and testing of BcWCL1 PASΔ regions(library_design)
This stage exists to determine which region of BcWCL1 PASΔ is responsible for transcriptional activation.
Selection: Different fragments from the N-terminal and C-terminal regions of BcWCL1 PASΔ were assessed for transcriptional activity.
- 2.Reporter-based functional characterization(functional_characterization)
This stage functionally tests whether BcWCL1 PASΔ fragments can drive transcription in yeast.
Selection: Transcriptional activity measured using chimeric transcription factors and the luciferase reporter gene.
- 3.Comparison to standard activation domains(secondary_characterization)
This stage benchmarks the identified activation domain against commonly used eukaryotic activation domains.
Selection: Activation levels of the BcWCL1 PASΔ activation domain were compared with Gal4 and p65.
Steps
- 1.Assess different N-terminal and C-terminal fragments of BcWCL1 PASΔengineered photoreceptor construct being mapped by fragment analysis
To localize the region responsible for transcriptional activation within BcWCL1 PASΔ.
Fragment-level mapping is needed before assigning activation function to a specific region.
- 2.Measure fragment transcriptional activity using chimeric transcription factors and a luciferase reportertested construct fragments in reporter assay
To determine which BcWCL1 PASΔ region functions as a transcriptional activation domain.
Reporter-based testing follows fragment design because functional readout is required to identify the active region.
- 3.Compare the identified BcWCL1 PASΔ activation domain to Gal4 and p65 activation domainsactivation domain being benchmarked
To place the BcWCL1 PASΔ activation domain in the context of commonly used eukaryotic activation domains.
Benchmarking is informative after the activation domain has been identified.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Techniques
No technique tags yet.
Target processes
transcriptionImplementation Constraints
The tool is a PAS-domain-deleted BcWCL1 construct, indicating that domain deletion is central to its design. The cited work places its function in yeast and identifies the N-terminal region as necessary for self-dimerization and transcriptional activation, but the supplied evidence does not provide construct architecture, promoter context, or cofactor requirements.
The supplied evidence is limited to domain mapping and comparative transcriptional activation claims in yeast. Quantitative performance details, kinetics, dynamic range, reversibility, spectral properties, and validation outside yeast are not provided in the supplied evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2primary paper2023MED
Ranked Claims
The transcriptional activation levels of the BcWCL1 PASΔ activation domain are comparable to those of Gal4 and p65 activation domains.
Finally, we determined that the transcriptional activation levels of BcWCL1PASΔ AD are comparable to those obtained with commonly used ADs in eukaryotic cells (Gal4 and p65).
The transcriptional activation levels of the BcWCL1 PASΔ activation domain are comparable to those of Gal4 and p65 activation domains.
Finally, we determined that the transcriptional activation levels of BcWCL1PASΔ AD are comparable to those obtained with commonly used ADs in eukaryotic cells (Gal4 and p65).
The transcriptional activation levels of the BcWCL1 PASΔ activation domain are comparable to those of Gal4 and p65 activation domains.
Finally, we determined that the transcriptional activation levels of BcWCL1PASΔ AD are comparable to those obtained with commonly used ADs in eukaryotic cells (Gal4 and p65).
The transcriptional activation levels of the BcWCL1 PASΔ activation domain are comparable to those of Gal4 and p65 activation domains.
Finally, we determined that the transcriptional activation levels of BcWCL1PASΔ AD are comparable to those obtained with commonly used ADs in eukaryotic cells (Gal4 and p65).
The transcriptional activation levels of the BcWCL1 PASΔ activation domain are comparable to those of Gal4 and p65 activation domains.
Finally, we determined that the transcriptional activation levels of BcWCL1PASΔ AD are comparable to those obtained with commonly used ADs in eukaryotic cells (Gal4 and p65).
The transcriptional activation levels of the BcWCL1 PASΔ activation domain are comparable to those of Gal4 and p65 activation domains.
Finally, we determined that the transcriptional activation levels of BcWCL1PASΔ AD are comparable to those obtained with commonly used ADs in eukaryotic cells (Gal4 and p65).
The transcriptional activation levels of the BcWCL1 PASΔ activation domain are comparable to those of Gal4 and p65 activation domains.
Finally, we determined that the transcriptional activation levels of BcWCL1PASΔ AD are comparable to those obtained with commonly used ADs in eukaryotic cells (Gal4 and p65).
The transcriptional activation levels of the BcWCL1 PASΔ activation domain are comparable to those of Gal4 and p65 activation domains.
Finally, we determined that the transcriptional activation levels of BcWCL1PASΔ AD are comparable to those obtained with commonly used ADs in eukaryotic cells (Gal4 and p65).
The N-terminal region of BcWCL1 PASΔ contains a functional transcriptional activation domain belonging to the 9aaTAD family.
By using chimeric transcription factors and the luciferase reporter gene, we assessed the transcriptional activity of different fragments of the N-terminal and C-terminal regions of BcWCL1PASΔ, identifying a functional transcriptional activation domain (AD) in the N-terminal region that belongs to the 9aaTAD family.
The N-terminal region of BcWCL1 PASΔ contains a functional transcriptional activation domain belonging to the 9aaTAD family.
By using chimeric transcription factors and the luciferase reporter gene, we assessed the transcriptional activity of different fragments of the N-terminal and C-terminal regions of BcWCL1PASΔ, identifying a functional transcriptional activation domain (AD) in the N-terminal region that belongs to the 9aaTAD family.
The N-terminal region of BcWCL1 PASΔ contains a functional transcriptional activation domain belonging to the 9aaTAD family.
By using chimeric transcription factors and the luciferase reporter gene, we assessed the transcriptional activity of different fragments of the N-terminal and C-terminal regions of BcWCL1PASΔ, identifying a functional transcriptional activation domain (AD) in the N-terminal region that belongs to the 9aaTAD family.
The N-terminal region of BcWCL1 PASΔ contains a functional transcriptional activation domain belonging to the 9aaTAD family.
By using chimeric transcription factors and the luciferase reporter gene, we assessed the transcriptional activity of different fragments of the N-terminal and C-terminal regions of BcWCL1PASΔ, identifying a functional transcriptional activation domain (AD) in the N-terminal region that belongs to the 9aaTAD family.
The N-terminal region of BcWCL1 PASΔ contains a functional transcriptional activation domain belonging to the 9aaTAD family.
By using chimeric transcription factors and the luciferase reporter gene, we assessed the transcriptional activity of different fragments of the N-terminal and C-terminal regions of BcWCL1PASΔ, identifying a functional transcriptional activation domain (AD) in the N-terminal region that belongs to the 9aaTAD family.
The N-terminal region of BcWCL1 PASΔ contains a functional transcriptional activation domain belonging to the 9aaTAD family.
By using chimeric transcription factors and the luciferase reporter gene, we assessed the transcriptional activity of different fragments of the N-terminal and C-terminal regions of BcWCL1PASΔ, identifying a functional transcriptional activation domain (AD) in the N-terminal region that belongs to the 9aaTAD family.
The N-terminal region of BcWCL1 PASΔ contains a functional transcriptional activation domain belonging to the 9aaTAD family.
By using chimeric transcription factors and the luciferase reporter gene, we assessed the transcriptional activity of different fragments of the N-terminal and C-terminal regions of BcWCL1PASΔ, identifying a functional transcriptional activation domain (AD) in the N-terminal region that belongs to the 9aaTAD family.
The N-terminal region of BcWCL1 PASΔ contains a functional transcriptional activation domain belonging to the 9aaTAD family.
By using chimeric transcription factors and the luciferase reporter gene, we assessed the transcriptional activity of different fragments of the N-terminal and C-terminal regions of BcWCL1PASΔ, identifying a functional transcriptional activation domain (AD) in the N-terminal region that belongs to the 9aaTAD family.
BcWCL1 PASΔ enables blue-light-dependent transcriptional activation as a single component in yeast.
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
BcWCL1 PASΔ enables blue-light-dependent transcriptional activation as a single component in yeast.
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
BcWCL1 PASΔ enables blue-light-dependent transcriptional activation as a single component in yeast.
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
BcWCL1 PASΔ enables blue-light-dependent transcriptional activation as a single component in yeast.
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
BcWCL1 PASΔ enables blue-light-dependent transcriptional activation as a single component in yeast.
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
BcWCL1 PASΔ enables blue-light-dependent transcriptional activation as a single component in yeast.
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
BcWCL1 PASΔ enables blue-light-dependent transcriptional activation as a single component in yeast.
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
BcWCL1 PASΔ enables blue-light-dependent transcriptional activation as a single component in yeast.
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
BcWCL1 PASΔ self-dimerizes through its N-terminal region upon blue-light stimulation.
Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
BcWCL1 PASΔ self-dimerizes through its N-terminal region upon blue-light stimulation.
Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
BcWCL1 PASΔ self-dimerizes through its N-terminal region upon blue-light stimulation.
Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
BcWCL1 PASΔ self-dimerizes through its N-terminal region upon blue-light stimulation.
Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
BcWCL1 PASΔ self-dimerizes through its N-terminal region upon blue-light stimulation.
Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
BcWCL1 PASΔ self-dimerizes through its N-terminal region upon blue-light stimulation.
Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
BcWCL1 PASΔ self-dimerizes through its N-terminal region upon blue-light stimulation.
Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
BcWCL1 PASΔ self-dimerizes through its N-terminal region upon blue-light stimulation.
Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
BcWCL1PASΔ shows a blue-light response and interacts with BcWCL2 or BcWCL2PASΔ upon light stimulation.
the BcWCL1<PASΔ> protein shows a blue-light response and interacts with BcWCL2 or BcWCL2<PASΔ> upon light stimulation
BcWCL1PASΔ shows a blue-light response and interacts with BcWCL2 or BcWCL2PASΔ upon light stimulation.
the BcWCL1<PASΔ> protein shows a blue-light response and interacts with BcWCL2 or BcWCL2<PASΔ> upon light stimulation
BcWCL1PASΔ shows a blue-light response and interacts with BcWCL2 or BcWCL2PASΔ upon light stimulation.
the BcWCL1<PASΔ> protein shows a blue-light response and interacts with BcWCL2 or BcWCL2<PASΔ> upon light stimulation
BcWCL1PASΔ shows a blue-light response and interacts with BcWCL2 or BcWCL2PASΔ upon light stimulation.
the BcWCL1<PASΔ> protein shows a blue-light response and interacts with BcWCL2 or BcWCL2<PASΔ> upon light stimulation
BcWCL1PASΔ shows a blue-light response and interacts with BcWCL2 or BcWCL2PASΔ upon light stimulation.
the BcWCL1<PASΔ> protein shows a blue-light response and interacts with BcWCL2 or BcWCL2<PASΔ> upon light stimulation
BcWCL1PASΔ shows a blue-light response and interacts with BcWCL2 or BcWCL2PASΔ upon light stimulation.
the BcWCL1<PASΔ> protein shows a blue-light response and interacts with BcWCL2 or BcWCL2<PASΔ> upon light stimulation
BcWCL1PASΔ shows a blue-light response and interacts with BcWCL2 or BcWCL2PASΔ upon light stimulation.
the BcWCL1<PASΔ> protein shows a blue-light response and interacts with BcWCL2 or BcWCL2<PASΔ> upon light stimulation
Both BcWCL1 and BcWCL1PASΔ are capable of light sensing.
we demonstrate that BcWCL1 and BcWCL1<PASΔ> respond to blue light by introducing a point mutation in the photoactive cysteine, confirming that both proteins are capable of light sensing
Both BcWCL1 and BcWCL1PASΔ are capable of light sensing.
we demonstrate that BcWCL1 and BcWCL1<PASΔ> respond to blue light by introducing a point mutation in the photoactive cysteine, confirming that both proteins are capable of light sensing
Both BcWCL1 and BcWCL1PASΔ are capable of light sensing.
we demonstrate that BcWCL1 and BcWCL1<PASΔ> respond to blue light by introducing a point mutation in the photoactive cysteine, confirming that both proteins are capable of light sensing
Both BcWCL1 and BcWCL1PASΔ are capable of light sensing.
we demonstrate that BcWCL1 and BcWCL1<PASΔ> respond to blue light by introducing a point mutation in the photoactive cysteine, confirming that both proteins are capable of light sensing
Both BcWCL1 and BcWCL1PASΔ are capable of light sensing.
we demonstrate that BcWCL1 and BcWCL1<PASΔ> respond to blue light by introducing a point mutation in the photoactive cysteine, confirming that both proteins are capable of light sensing
Both BcWCL1 and BcWCL1PASΔ are capable of light sensing.
we demonstrate that BcWCL1 and BcWCL1<PASΔ> respond to blue light by introducing a point mutation in the photoactive cysteine, confirming that both proteins are capable of light sensing
Both BcWCL1 and BcWCL1PASΔ are capable of light sensing.
we demonstrate that BcWCL1 and BcWCL1<PASΔ> respond to blue light by introducing a point mutation in the photoactive cysteine, confirming that both proteins are capable of light sensing
Deletion of the PAS domain in BcWCL1 or BcWCL2 severely impairs interaction between these proteins.
Deletion of the PAS domains present in BcWCL1 (BcWCL1<PASΔ>) or BcWCL2 (BcWCL2<PASΔ>) severely impairs the interaction between these proteins.
Deletion of the PAS domain in BcWCL1 or BcWCL2 severely impairs interaction between these proteins.
Deletion of the PAS domains present in BcWCL1 (BcWCL1<PASΔ>) or BcWCL2 (BcWCL2<PASΔ>) severely impairs the interaction between these proteins.
Deletion of the PAS domain in BcWCL1 or BcWCL2 severely impairs interaction between these proteins.
Deletion of the PAS domains present in BcWCL1 (BcWCL1<PASΔ>) or BcWCL2 (BcWCL2<PASΔ>) severely impairs the interaction between these proteins.
Deletion of the PAS domain in BcWCL1 or BcWCL2 severely impairs interaction between these proteins.
Deletion of the PAS domains present in BcWCL1 (BcWCL1<PASΔ>) or BcWCL2 (BcWCL2<PASΔ>) severely impairs the interaction between these proteins.
Deletion of the PAS domain in BcWCL1 or BcWCL2 severely impairs interaction between these proteins.
Deletion of the PAS domains present in BcWCL1 (BcWCL1<PASΔ>) or BcWCL2 (BcWCL2<PASΔ>) severely impairs the interaction between these proteins.
Deletion of the PAS domain in BcWCL1 or BcWCL2 severely impairs interaction between these proteins.
Deletion of the PAS domains present in BcWCL1 (BcWCL1<PASΔ>) or BcWCL2 (BcWCL2<PASΔ>) severely impairs the interaction between these proteins.
Deletion of the PAS domain in BcWCL1 or BcWCL2 severely impairs interaction between these proteins.
Deletion of the PAS domains present in BcWCL1 (BcWCL1<PASΔ>) or BcWCL2 (BcWCL2<PASΔ>) severely impairs the interaction between these proteins.
Approval Evidence
2 sources7 linked approval claimsfirst-pass slugs bcwcl1-pas, bcwcl1pas
Recently, the blue-light response of BcWCL1 was demonstrated in a version without PAS domains (BcWCL1PASΔ). Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
Source:
Deletion of the PAS domains present in BcWCL1 (BcWCL1PAS) or BcWCL2 (BcWCL2PAS) severely impairs the interaction between these proteins.
Source:
comparisonsupports
The transcriptional activation levels of the BcWCL1 PASΔ activation domain are comparable to those of Gal4 and p65 activation domains.
Finally, we determined that the transcriptional activation levels of BcWCL1PASΔ AD are comparable to those obtained with commonly used ADs in eukaryotic cells (Gal4 and p65).
Source:
domain mappingsupports
The N-terminal region of BcWCL1 PASΔ contains a functional transcriptional activation domain belonging to the 9aaTAD family.
By using chimeric transcription factors and the luciferase reporter gene, we assessed the transcriptional activity of different fragments of the N-terminal and C-terminal regions of BcWCL1PASΔ, identifying a functional transcriptional activation domain (AD) in the N-terminal region that belongs to the 9aaTAD family.
Source:
functionsupports
BcWCL1 PASΔ enables blue-light-dependent transcriptional activation as a single component in yeast.
Interestingly, we observed that BcWCL1PASΔ enables transcriptional activation as a single component in yeast.
Source:
mechanismsupports
BcWCL1 PASΔ self-dimerizes through its N-terminal region upon blue-light stimulation.
Here, we demonstrated that BcWCL1PASΔ is capable of self-dimerization through its N-terminal region upon blue-light stimulation.
Source:
light responsivenesssupports
BcWCL1PASΔ shows a blue-light response and interacts with BcWCL2 or BcWCL2PASΔ upon light stimulation.
the BcWCL1<PASΔ> protein shows a blue-light response and interacts with BcWCL2 or BcWCL2<PASΔ> upon light stimulation
Source:
light sensingsupports
Both BcWCL1 and BcWCL1PASΔ are capable of light sensing.
we demonstrate that BcWCL1 and BcWCL1<PASΔ> respond to blue light by introducing a point mutation in the photoactive cysteine, confirming that both proteins are capable of light sensing
Source:
mutation effectsupports
Deletion of the PAS domain in BcWCL1 or BcWCL2 severely impairs interaction between these proteins.
Deletion of the PAS domains present in BcWCL1 (BcWCL1<PASΔ>) or BcWCL2 (BcWCL2<PASΔ>) severely impairs the interaction between these proteins.
Source:
Comparisons
Source-backed strengths
The N-terminal region of BcWCL1 PASΔ was mapped as a functional transcriptional activation domain of the 9aaTAD family. In the cited yeast study, the transcriptional activation levels of the BcWCL1 PASΔ activation domain were comparable to those of the Gal4 and p65 activation domains.
Ranked Citations
- 1.
- 2.