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Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development
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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
PubMed

Insights

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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