Chemical inhibition of stomatal differentiation by perturbation of the master-regulatory bHLH heterodimer via an ACT-Like domain
Selective perturbation of protein interactions with chemical compounds enables dissection and control of developmental processes. Differentiation of stomata, cellular valves vital for plant growth and survival, is specified by the basic-helix-loop-helix (bHLH) heterodimers. Harnessing a new amination reaction, we here report a synthesis, derivatization, target identification, and mode of action of an atypical doubly-sulfonylated imidazolone, Stomidazolone, which triggers stomatal stem cell arrest. Our forward chemical genetics followed by biophysical analyses elucidates that Stomidazolone directly binds to the C-terminal ACT-Like (ACTL) domain of MUTE, a master regulator of stomatal differentiation, and perturbs its heterodimerization with a partner bHLH, SCREAM in vitro and in plant cells. On the other hand, Stomidazolone analogs that are biologically inactive do not bind to MUTE or disrupt the SCREAM-MUTE heterodimers. Guided by structural docking modeling, we rationally design MUTE with reduced Stomidazolone binding. These engineered MUTE proteins are fully functional and confer Stomidazolone resistance in vivo. Our study identifies doubly-sulfonylated imidazolone as a direct inhibitor of the stomatal master regulator, further expanding the chemical space for perturbing bHLH-ACTL proteins to manipulate plant development.
Authors
- Soon‐Ki Han (ORCID: https://orcid.org/0000-0003-0890-4680)
- Kota Hashimoto (ORCID: https://orcid.org/0000-0002-7950-9947)
- Keiko U. Torii (ORCID: https://orcid.org/0000-0002-6168-427X)
- Krishna Mohan Sepuru (ORCID: https://orcid.org/0000-0002-3675-6294)
- Calvin M. Coffin (ORCID: https://orcid.org/0009-0007-2154-791X)
- Hiroe Kato
- Rie Iwasaki
- Toshinori Kinoshita (ORCID: https://orcid.org/0000-0001-7621-1259)
- Shu Jan Yip
- Ayami Nakagawa (ORCID: https://orcid.org/0000-0001-7230-9451)
- Daisuke Kurihara (ORCID: https://orcid.org/0000-0003-2703-0405)
- Kei Murakami (ORCID: https://orcid.org/0000-0001-5405-3069)
- Yasutomo Segawa (ORCID: https://orcid.org/0000-0001-6439-8546)
- Hyemin Seo (ORCID: https://orcid.org/0000-0002-2154-1151)
- Kenichiro Itami (ORCID: https://orcid.org/0000-0001-5227-7894)
- Stephanie Kim (ORCID: https://orcid.org/0000-0003-1245-9006)
- Zixuan Li (ORCID: https://orcid.org/0000-0003-2488-814X)
- Yusuke Aihara (ORCID: https://orcid.org/0000-0002-4081-3802)
Institutions
- Howard Hughes Medical Institute (US)
- The Graduate University for Advanced Studies, SOKENDAI (JP)
- Kwansei Gakuin University (JP)
- Japan Science and Technology Agency (JP)
- Institute for Molecular Science (JP)
- Nagoya University (JP)
- The University of Texas at Austin (US)
Publication Details
- Journal
- Nature Communications
- Published
- 2024-10-23
- DOI
- https://doi.org/10.1038/s41467-024-53214-4
- Citations
- 6
- Primary Topic
- Plant Gene Expression Analysis
- Type
- article
- Field-Weighted Citation Impact
- 1.40
Funders
- Howard Hughes Medical Institute
- University of Texas at Austin
- Tatematsu Foundation
- Ministry of Education, Culture, Sports, Science and Technology
- TOBE MAKI Scholarship Foundation
- Japan Society for the Promotion of Science
- Japan Science and Technology Agency
- Core Research for Evolutional Science and Technology
- Precursory Research for Embryonic Science and Technology