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Investigation of cellular signaling and epigenetic dynamics via optogenetic control of nuclear cytoplasmic distribution
Author(s) -
Yumerefendi Hayretin,
Kuhlman Brian
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.763.5
Subject(s) - microbiology and biotechnology , transcription factor , nuclear transport , chromatin , cytoplasm , biology , yeast , nuclear protein , nuclear export signal , nuclear localization sequence , ubiquitin , chemistry , cell nucleus , biochemistry , dna , gene
The ability to control nuclear cytoplasmic distribution of proteins in vivo with spatial and temporal precision would allow us to probe the functions of a broad range of proteins and uncouple their roles in cellular signaling. Here we report achieving reversible, blue light‐dependent control of nuclear import and nuclear export by engineering a nuclear localization and export signals into the Light Oxygen Voltage (LOV2) domain of phototropin 1 from Avena sativa . We validated predictions first in mammalian cells and in yeast. The Light Activated Nuclear Shuttle (LANS) and the Light Induced Nuclear eXporter (LINX) interact with the endogenous nucleocytoplasmic transport machinery for rapid nuclear import and export in mammalian cells, yeast and C. elegans embryos. In yeast, LANS allowed robust control of transcriptional activation via light‐induced nuclear accumulation of a transcription factor demonstrating its potential to activate nuclear proteins. Similarly, we demonstrated the use of LINX for transcriptional repression in yeast by ejecting a transcription factor out of the nucleus. Moreover, fusing the chromatin modifier Bre1 to LINX, we achieved light‐dependent control of histon H2B monoubiquitylation in yeast, revealing fast turnover of the ubiquitin mark. This inducible system allowed us to dynamically monitor the status of epigenetic modifications dependent on H2B ubiquitylation.