Control of Protein Activity and Cell Fate Specification via Light-Mediated Nuclear Translocation
Hayretin Yumerefendi1, Daniel J Dickinson2, Hui Wang3
1Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina, United States of America.
Plos One
|June 18, 2015
Summary
Researchers developed a light-activated nuclear shuttle (LANS) for precise control of protein localization. This tool enables light-dependent gene transcription and cell fate manipulation in vivo, offering new possibilities for biological research.
Area of Science:
- Molecular Biology
- Genetics
- Developmental Biology
Background:
- Light-activatable proteins offer spatiotemporal control over biological processes.
- The Light Oxygen Voltage (LOV) domain, specifically AsLOV2, can inhibit a nuclear localization signal (NLS) in its dark-state conformation.
- Previous studies demonstrated LOV/NLS control of protein localization in mammalian cells.
Purpose of the Study:
- To characterize the binding properties of a LOV/NLS switch.
- To demonstrate the utility of this switch, termed LANS (light-activated nuclear shuttle), for controlling gene transcription in yeast.
- To validate LANS function in vivo, specifically in C. elegans embryos, for controlling nuclear localization and cell fate.
Main Methods:
- Characterization of LOV/NLS switch binding properties.
- Implementation of LANS for light-induced gene transcription control in yeast.
- In vivo testing of LANS in C. elegans embryos for cellular nuclear localization control.
- Integration of LANS into the C. elegans lin-1 locus via Cas9-triggered homologous recombination to control cell fate.
Main Results:
- The LOV/NLS switch, LANS, was characterized for its binding properties.
- LANS successfully controlled gene transcription in yeast upon blue light activation.
- LANS enabled light-dependent control of nuclear localization in individual C. elegans embryonic cells.
- Light-dependent manipulation of a native transcription factor via LANS insertion into the lin-1 locus resulted in control of cell fate.
Conclusions:
- LANS functions effectively in yeast and C. elegans embryos.
- The LANS system provides spatial and temporal control over nuclear localization in vivo.
- LANS represents a valuable experimental tool for precise manipulation of biological processes in living organisms.
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