Evidence for functionality of transmembrane domains of predicted non-canonical plant phosphotransmitters
- Authors: Lomin S.N.1, Savelieva E.M.1, Elanskaya A.S.1, Arkhipov D.V.1, Romanov G.A.1
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Affiliations:
- Timiryazev Institute of Plant Physiology of the Russian Academy of Sciences
- Issue: Vol 522, No 1 (2025)
- Pages: 358-363
- Section: Articles
- URL: https://jdigitaldiagnostics.com/2686-7389/article/view/686381
- DOI: https://doi.org/10.31857/S2686738925030079
- ID: 686381
Cite item
Abstract
Bioinformatic methods have been used to predict a new subclass of proteins among plant phosphotransmitters involved in the signaling system of multistep phosphorelay. In contrast to the canonical soluble nucleocytosolic forms, the found phosphotransmitter sequences, belonging to a wide range of plant taxa, potentially contain transmembrane domains. This suggests localization of such proteins on cell membranes and, therefore, a different function in signaling than that of canonical phosphotransmitters. We tested the functionality of the transmembrane domain of the phosphotransmitter using the protein of the tea plant Camellia sinensis. Membrane localization of the transiently expressed recombinant phosphotransmitter with this domain was confirmed by microscopy and immunoblotting. Thus, for the first time, experimental evidence was obtained for the existence of membrane-bound plant phosphotransmitters with as yet unknown functions. These data suggest the presence of a non-canonical membrane branch of signal transduction in the multistage phosphotransfer system in plants.
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About the authors
S. N. Lomin
Timiryazev Institute of Plant Physiology of the Russian Academy of Sciences
Email: savelievaek@ya.ru
Russian Federation, Moscow
E. M. Savelieva
Timiryazev Institute of Plant Physiology of the Russian Academy of Sciences
Author for correspondence.
Email: savelievaek@ya.ru
Russian Federation, Moscow
A. S. Elanskaya
Timiryazev Institute of Plant Physiology of the Russian Academy of Sciences
Email: savelievaek@ya.ru
Russian Federation, Moscow
D. V. Arkhipov
Timiryazev Institute of Plant Physiology of the Russian Academy of Sciences
Email: savelievaek@ya.ru
Russian Federation, Moscow
G. A. Romanov
Timiryazev Institute of Plant Physiology of the Russian Academy of Sciences
Email: savelievaek@ya.ru
Russian Federation, Moscow
References
- Mira-Rodado V. New Insights into Multistep-Phosphorelay (MSP)/Two-Component System (TCS) Regulation: Are Plants and Bacteria That Different? // Plants. 2019. № 8. Р. 590.
- Argueso C.T., Kieber J.J. Cytokinin: From autoclaved DNA to two-component signaling // Plant Cell. 2024. № 36. Р. 1429.
- Huo R., Liu Z., Yu X., Li Z. The Interaction Network and Signaling Specificity of Two-Component System in Arabidopsis // International Journal of Molecular Sciences. 2020. 21(14):4898. https://doi.org/10.3390/ijms21144898
- Arkhipov D.V., Lomin S.N., Myakushina Y.A., et al. Modeling of protein–protein interactions in cytokinin signal transduction // Int. J. Mol. Sci. 2019. 20:2096.
- Schaller G.E., Bishopp A., Kieber J.J. The yin-yang of hormones: cytokinin and auxin interactions in plant development // Plant Cell. 2015. 27:44.
- Králová M., Kubalová I., Hajný J., et al. A decoy receptor derived from alternative splicing fine-tunes cytokinin signaling in Arabidopsis // Mol Plant. 2024. 17:1850.
- Suzuki T., Sakurai K., Imamura A., et al. Compilation and characterization of histidine-containing phosphotransmitters implicated in His-to-Asp phosphorelay in plants: AHP signal transducers of Arabidopsis thaliana // Biosci. Biotechnol. Biochem. 2000. 64:2486.
- Ito Y., Kurata N. Identification and characterization of cytokinin-signalling gene families in rice // Gene. 2006. 382:57.
- Punwani J.A., Kieber J.J. Localization of the Arabidopsis histidine phosphotransfer proteins is independent of cytokinin // Plant Signal. Behav. 2010. 5:896.
- Yuan L., Liu Z., Song X., Johnson C., Yu X., Sundaresan V. The CKI1 histidine kinase specifies the female gametic precursor of the endosperm // Dev Cell. 2016. 37:34.
- Lomin S.N., Yonekura-Sakakibara K., Romanov G.A., Sakakibara H. Ligand-binding properties and subcellular localization of maize cytokinin receptors // J Exp Bot. 2011. 62:5149.
- Zhang H., Linster E., Gannon L., Leemhuis W., Rundle C.A., Theodoulou F.L., Wirtz M. Tandem fluorescent protein timers for noninvasive relative protein lifetime measurement in plants // Plant Physiol. 2019. 180:718.
- Karimi M., De Meyer B., Hilson P. Modular cloning in plant cells // Trends Plant Sci. 2005. 10:103.
- Sparkes I.A., Runions J., Kearns A., Hawes C. Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants // Nat. Protoc. 2006. 1:2019.
- Savelieva E.M., Lomin S.N., Romanov G.A. A modified method for quantification of cytokinin-receptor binding using isolated plant microsomes enriched with cognate transmembrane receptors // Russ. J. Plant Physiol. 2022. 69:6.
- Lomin S.N., Romanov G.A. A new tool for quantification of membrane protein partitioning between different cell membranes // Anal Biochem. 2020. 599:113734.
- Lomin S.N., Myakushina Y.A., Arkhipov D.V., Leonova O.G., Popenko V.I., Schmülling T., Romanov G.A. Studies of cytokinin receptor-phosphotransmitter interaction provide evidences for the initiation of cytokinin signalling in the endoplasmic reticulum // Funct Plant Biol. 2018. 45(2):192-202.
- Li K., Li H., Liang W.L., Liu J.J., Tian H.Y., Wang L.H., Wei Y.H. Identification of the AHP family reveals their critical response to cytokinin regulation during adventitious root formation in apple rootstock // Front Plant Sci. 2025. 15:1511713.
Supplementary files

Note
Presented by Academician of the RAS S.G. Georgieva