Light generation of intracellular Ca2+ signals by a genetically encoded protein BACCS

Tomohiro Ishii, Koji Sato, Toshiyuki Kakumoto, Shigenori Miura, Kazushige Touhara, Shoji Takeuchi and Takao Nakata ()
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Tomohiro Ishii: Graduate School of Medical and Dental Science, Tokyo Medical and Dental University
Koji Sato: Okazaki Institute for Integrative Bioscience
Toshiyuki Kakumoto: Graduate School of Medical and Dental Science, Tokyo Medical and Dental University
Shigenori Miura: Institute of Industrial Science, The University of Tokyo
Kazushige Touhara: Graduate School of Agricultural and Life Sciences, The University of Tokyo
Shoji Takeuchi: Institute of Industrial Science, The University of Tokyo
Takao Nakata: Graduate School of Medical and Dental Science, Tokyo Medical and Dental University

Nature Communications, 2015, vol. 6, issue 1, 1-15

Abstract: Abstract Ca2+ signals are highly regulated in a spatiotemporal manner in numerous cellular physiological events. Here we report a genetically engineered blue light-activated Ca2+ channel switch (BACCS), as an optogenetic tool for generating Ca2+ signals. BACCS opens Ca2+-selective ORAI ion channels in response to light. A BACCS variant, dmBACCS2, combined with Drosophila Orai, elevates the Ca2+ concentration more rapidly, such that Ca2+ elevation in mammalian cells is observed within 1 s on light exposure. Using BACCSs, we successfully control cellular events including NFAT-mediated gene expression. In the mouse olfactory system, BACCS mediates light-dependent electrophysiological responses. Furthermore, we generate BACCS mutants, which exhibit fast and slow recovery of intracellular Ca2+. Thus, BACCSs are a useful optogenetic tool for generating temporally various intracellular Ca2+ signals with a large dynamic range, and will be applicable to both in vitro and in vivo studies.

Date: 2015
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DOI: 10.1038/ncomms9021

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