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Lomin, S. N.

Publications and source records attributed to Lomin, S. N..

2 recordsLinked to original sources

Cytokinin perception in potato: New features of canonic players

Potato is the most economically important non-cereal food crop. Tuber formation in potato is regulated by phytohormones, cytokinins (CKs) in particular. The present work was aimed to study CK signal perception in potato. The sequenced potato genome of doubled monoploid Phureja was used for bioinformatic analysis and as a tool for identification of putative CK receptors from autotetraploid potato cv. Desiree. All basic elements of multistep phosphorelay (MSP) required for CK signal transduction were identified in Phureja genome, including three genes orthologous to three CK receptor genes (AHK 2-4) of Arabidopsis. As distinct from Phureja, autotetraploid potato contains at least two allelic isoforms of each receptor type. Putative receptor genes from Desiree plants were cloned, sequenced and expressed, and main characteristics of encoded proteins, firstly their consensus motifs, structure models, ligand-binding properties, and the ability to transmit CK signal, were determined. In all studied aspects the predicted sensor histidine kinases met the requirements for genuine CK receptors. Expression of potato CK receptors was found to be organ-specific and sensitive to growth conditions, particularly to sucrose content. Our results provide a solid basis for further in-depth study of CK signaling system and biotechnological improvement of potato.

plant biology

Cytokinin activity of N6-benzyladenine derivatives assayed by interaction with the receptors in planta, in vitro, and in silico

Biological effects of hormones in both plants and animals are based on high-affrnity interaction with cognate receptors resulting in their activation. The signal of cytokinins, classical plant hormones, is perceived in Arabidopsis by three homologous membrane receptors: AHK2, AHK3, and CRE1/AHK4. To study the cytokinin-receptor interaction, we used 25 derivatives of potent cytokinin N6-benzyladenine (BA) with substituents in the purine heterocycle and/or in the side chain. The study was focused primarily on individual cytokinin receptors from Arabidopsis. The main in planta assay system was based on Arabidopsis double mutants retaining only one isoform of cytokinin receptors and harboring cytokinin-sensitive reporter gene. Classical cytokinin biotest with Amaranthus seedlings was used as an additional biotest. In parallel, the binding of ligands to individual cytokinin receptors was assessed in the in vitro test system. Quantitative comparison of results of different assays confirmed the partial similarity of ligand-binding properties of receptor isoforms. Substituents at positions 8 and 9 of adenine moiety, elongated linker up to 4 methylene units, replacement of N6 by sulfur or oxygen, resulted in suppression of cytokinin activity of the derivative towards all receptors. Introduction of a halogen into position 2 of adenine moiety, on the contrary, often increased the ligand activity, especially toward AHK3. Features both common and distinctive of cytokinin receptors in Arabidopsis and Amaranthus were revealed, highlighting species specificity of the cytokinin perception apparatus. Correlations between extent of compound binding to a receptor in vitro and its ability to activate the same receptor in planta were evaluated for each AHK protein. Interaction patterns between individual receptors and ligands were rationalized by structure analysis and molecular docking in sensory modules of AHK receptors. The best correlation between docking scores and specific binding was observed for AHK3. In addition, receptor-specific ligands have been discovered with unique properties to predominantly activate or block distinct cytokinin receptors. These ligands are promising for practical application and as molecular tools in the study of the cytokinin perception by plant cells.\n\nGraphical abstract\n\nO_FIG O_LINKSMALLFIG WIDTH=177 HEIGHT=200 SRC=\"FIGDIR/small/241281_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (85K):\norg.highwire.dtl.DTLVardef@1fef3f9org.highwire.dtl.DTLVardef@143a10eorg.highwire.dtl.DTLVardef@102d62corg.highwire.dtl.DTLVardef@13b1db1_HPS_FORMAT_FIGEXP M_FIG C_FIG Individual cytokinin receptors from Arabidopsis were assayed in planta, in vitro and in silico with 25 different 6-benzyladenine derivatives, new receptor-specific cytokinins were revealed.

plant biology