An Optogenetic Tool for Induced Protein Stabilization Based on the Phaeodactylum tricornutum Aureochrome 1a Light–Oxygen–Voltage Domain
| Autor: | Christof Taxis, Sebastian Hepp, Lars-Oliver Essen, Sophia Hasenjäger, Jonathan Trauth, Filipp Bezold |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Photoreceptors
Plant Light Protein Conformation Saccharomyces cerevisiae Protein degradation Optogenetics Adenylyl cyclase 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Protein Domains Structural Biology Cyclic adenosine monophosphate Phaeodactylum tricornutum Molecular Biology 030304 developmental biology Diatoms 0303 health sciences biology Protein Stability Rational design biology.organism_classification Oxygen chemistry Biophysics Protein stabilization 030217 neurology & neurosurgery Transcription Factors |
| Zdroj: | Journal of Molecular Biology. 432:1880-1900 |
| ISSN: | 0022-2836 |
| DOI: | 10.1016/j.jmb.2020.02.019 |
| Popis: | Control of cellular events by optogenetic tools is a powerful approach to manipulate cellular functions in a minimally invasive manner. A common problem posed by the application of optogenetic tools is to tune the activity range to be physiologically relevant. Here, we characterized a photoreceptor of the light-oxygen-voltage (LOV) domain family of Phaeodactylum tricornutum aureochrome 1a (AuLOV) as a tool for increasing protein stability under blue light conditions in budding yeast. Structural studies of AuLOVwt, the variants AuLOVM254, and AuLOVW349 revealed alternative dimer association modes for the dark state, which differ from previously reported AuLOV dark-state structures. Rational design of AuLOV-dimer interface mutations resulted in an optimized optogenetic tool that we fused to the photoactivatable adenylyl cyclase from Beggiatoa sp. This synergistic light-regulation approach using two photoreceptors resulted in an optimized, photoactivatable adenylyl cyclase with a cyclic adenosine monophosphate production activity that matches the physiological range of Saccharomyces cerevisiae. Overall, we enlarged the optogenetic toolbox for yeast and demonstrated the importance of fine-tuning the optogenetic tool activity for successful application in cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect