@article{b6bbd5a2b75b4bdda6cfbad34ee4db2e,
title = "Rational design and implementation of a chemically inducible heterotrimerization system",
abstract = "Chemically inducible dimerization (CID) uses a small molecule to induce binding of two different proteins. CID tools such as the FK506-binding protein–FKBP–rapamycin-binding– (FKBP–FRB)–rapamycin system have been widely used to probe molecular events inside and outside cells. While various CID tools are available, chemically inducible trimerization (CIT) does not exist, due to inherent challenges in designing a chemical that simultaneously binds three proteins with high affinity and specificity. Here, we developed CIT by rationally splitting FRB and FKBP. Cellular and structural datasets showed efficient trimerization of split pairs of FRB or FKBP with full-length FKBP or FRB, respectively, by rapamycin. CIT rapidly induced tri-organellar junctions and perturbed intended membrane lipids exclusively at select membrane contact sites. By conferring one additional condition to what is achievable with CID, CIT expands the types of manipulation in single live cells to address cell biology questions otherwise intractable and engineer cell functions for future synthetic biology applications.",
author = "Wu, {Helen D.} and Masaki Kikuchi and Onur Dagliyan and Aragaki, {Adam K.} and Hideki Nakamura and Dokholyan, {Nikolay V.} and Takashi Umehara and Takanari Inoue",
note = "Funding Information: We thank L. Bertozzi and S. Thompson for help with plasmid generation. We thank R. DeRose, X.Y. Zhou and Y. Nihongaki for proofreading the manuscript. We thank H. Niwa (RIKEN), N. Sakai (RIKEN) and the staff at the BL26B2 beamline (Proposal No. 20190047) of SPring-8 (Harima, Japan) and the X06DA beamline (Proposal No. 20171001) of the Swiss Light Source, Paul Scherrer Institut (Villigen, Switzerland) for their help in X-ray diffraction data collection. We acknowledge support from the National Institutes for Health (grant nos. 5R01GM123130 to T.I., and 5R01GM123247 and 1R35 GM134864 to N.V.D.), the Passan Foundation to N.V.D., the DoD DARPA (grant no. HR0011-16-C-0139 to T.I.), and the PRESTO program of the Japan Science and Technology Agency to T.U. (grant no. JPMJPR12A3) and T.I. (grant no. JPMJPR12A5) and a Grant-in-Aid for Scientific Research (B) to T.U. (grant no. 16H05089) from the Japan Society for the Promotion of Science. Publisher Copyright: {\textcopyright} 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.",
year = "2020",
month = sep,
day = "1",
doi = "10.1038/s41592-020-0913-x",
language = "English (US)",
volume = "17",
pages = "928--936",
journal = "Nature Methods",
issn = "1548-7091",
publisher = "Nature Publishing Group",
number = "9",
}