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Kubalova, I.

Publications and source records attributed to Kubalova, I..

3 recordsLinked to original sources

Decoy Receptor Fine-tunes Cytokinin Signaling

Hormone perception and signaling pathways play a fundamental regulatory function in cell growth, developmental, and physiological processes in both plant and animal systems. Those pathways are activated by hormone binding to the receptor to trigger cellular responses. Equally important are mechanisms that suppress activated transduction cascades to reset the system. Different mechanisms at the level of hormone biosynthesis and deactivation through degradation, conjugation, and production of repressors that attenuate transduction cascades downstream of receptors are known. In animal systems, decoy receptors have been identified as another important mechanism for fine-tuning the activity of the signaling pathways in processes like inflammatory responses, apoptosis, and blood vessel formation. Decoy receptors recognize and bind specific signaling molecules, but they cannot activate downstream signaling pathways thus providing competitive inhibition. Here we describe the first decoy receptor in plants. We show that the splicing variant of CRE1/AHK4 receptor of cytokinin, a hormone with a key role in the regulation of cell division and meristem maintenance in plants, acts as a decoy receptor to attenuate cytokinin signaling. We propose that this novel mechanism of signaling control applies in processes when modulation of CK signaling is needed.

plant biology↗

Helical metaphase chromatid coiling is conserved

The higher-order organization of metaphase chromosomes has been debated for almost 140 years. Classical light and electron microscopy studies suggested that chromatids are composed of helically organized chromatin fibers (chromonemata). Non-helical models were also recently proposed. We studied chromosome organization in barley using cutting-edge approaches and obtained evidence for a helically arranged 400-nm chromatin fiber representing the chromonema within chromatid arms. The number of turns is positively correlated with arm length. Turn size and chromatin density decrease towards the telomeres. Due to their specialized functions, the helical organization of centromeres and nucleolus-organizing regions is interrupted by several thinner, straight chromatin fibers. A comparison with previously published data indicates that the helical turning of metaphase chromatid arms is a conserved feature of large eukaryotic chromosomes.

cell biology↗

Prospects and limitations of expansion microscopy in chromatin ultrastructure determination

Expansion Microscopy (ExM) is a method to magnify physically a specimen with preserved ultrastructure. It has the potential to explore structural features beyond the diffraction limit of light. The procedure has been successfully used for different animal species, from isolated macromolecular complexes through cells to tissue slices. Expansion of plant-derived samples is still at the beginning, and little is known whether the chromatin ultrastructure becomes altered by physical expansion. In this study, we expanded isolated barley nuclei and compared whether ExM can provide a structural view of chromatin comparable with super-resolution microscopy. Different fixation and denaturation/digestion conditions were tested to maintain the chromatin ultrastructure. We achieved up to [~]4.2-times physically expanded nuclei corresponding to a maximal resolution of [~]50-60 nm when imaged by wild-field (WF) microscopy. By applying structured illumination microscopy (SIM, super-resolution) doubling the WF resolution the chromatin structures were observed at a resolution of [~]25-35 nm. WF microscopy showed a preserved nucleus shape and nucleoli. Moreover, we were able to detect chromatin domains, invisible in unexpanded nuclei. However, by applying SIM we observed that the preservation of the chromatin ultrastructure after expansion was not complete and that the majority of the tested conditions failed to keep the ultrastructure. Nevertheless, using expanded nuclei we detected successfully centromere repeats by fluorescence in situ hybridization (FISH) and the centromere-specific histone H3 variant CENH3 by indirect immunostaining. However, although these repeats and proteins were localized at the correct position within the nuclei (indicating a Rabl orientation) their ultrastructural arrangement was impaired.

cell biology↗