Phytochromes function as red/far-red photoreceptors in plants and are essential for light-regulated growth development. Photomorphogenesis, the developmental program light, is default seed plants. In dark-grown seedlings, photomorphogenic suppressed by action of CONSTITUTIVELY PHOTOMORPHOGENIC1 (COP1)/SUPPRESSOR OF phyA-105 (SPA) complex, which targets positive regulators degradation proteasome. inhibit COP1/SPA leading to accumulation transcription factors promoting photomorphogenesis; yet,...

Intron splicing increases proteome complexity, promotes RNA stability, and enhances transcription. However, introns the concomitant need for extend time required gene expression can cause an undesirable delay in activation of genes. Here, we show that plant microRNA processing factor SERRATE (SE) plays unexpected pivotal role regulation intronless Arabidopsis SE associated with more than 1000, mainly intronless, genes a transcription-dependent manner. Chromatin-bound liaised paused...

As technical developments in omics and biomedical imaging increase the throughput of data generation life sciences, need for information systems capable managing heterogeneous digital assets is increasing. In particular, supporting findability, accessibility, interoperability, reusability (FAIR) principles scientific management.We propose a Service Oriented Architecture approach integrated management analysis multi-omics data. Our architecture introduces an image system into FAIR-supporting,...

The UV-A/blue light photoreceptor crytochrome2 (cry2) plays a fundamental role in the transition from vegetative to reproductive phase facultative long-day plant Arabidopsis thaliana. cry2 protein level strongly decreases when etiolated seedlings are exposed blue light; is first phosphorylated, polyubiquitinated, and then degraded by 26S proteasome. COP1 involved degradation, but several cop1 mutants show only reduced not abolished degradation. SUPPRESSOR OF PHYA-105 (SPA) proteins known...

Protein-protein interaction studies provide valuable insights into cellular signaling. Brassinosteroid (BR) signaling is initiated by the hormone-binding receptor Insensitive 1 (BRI1) and its co-receptor BRI1 Associated Kinase (BAK1). BAK1 were shown to interact independently with Receptor-Like Protein 44 (RLP44), which implicated in BRI1/BAK1-dependent cell wall integrity perception. To demonstrate proposed complex formation of BRI1, RLP44, we established three-fluorophore intensity-based...

Super-resolution microscopy (SRM) approaches revolutionize cell biology by providing insights into the nanoscale organization and dynamics of macromolecular assemblies single molecules in living cells. A major hurdle limiting SRM democratization is post-acquisition data analysis which often complex time-consuming. Here, we present OneFlowTraX, a user-friendly open-source software dedicated to single-molecule localization (SMLM) such as single-particle tracking photoactivated (sptPALM)....

The distribution of proteins across the plasma membrane is not uniform; however, principles governing their organization remain fully understood. Hypersensitive-induced reaction (HIR) are plant-specific members stomatin/prohibitin/flotillin/HflK/C (SPFH) family that have been shown to influence organization. Arabidopsis thaliana HIR2 interacts with multiple proteins, including receptor kinases such as BAK1-INTERACTING RECEPTORS 2 and 3 (BIR2 3), BRI1-ASSOCIATED KINASE 1 (BAK1), FLAGELLIN...

Glioblastoma WHO IV belongs to a group of brain tumors that are still incurable. A promising treatment approach applies photodynamic therapy (PDT) with hypericin as photosensitizer. To generate comprehensive understanding the photosensitizer-tumor interactions, first part our study is focused on investigating distribution and penetration behavior in glioma cell spheroids by fluorescence microscopy. In second part, lifetime imaging microscopy (FLIM) was used correlate (FLT) changes...

Hypericin is one of the most efficient photosensitizers used in photodynamic tumor therapy (PDT). The reported treatments this drug reach from antidepressive, antineoplastic, antitumor and antiviral activity. We show that hypericin can be optically detected down to a single molecule at ambient conditions. even observed inside cancer cell, which implies directly for advanced microscopy techniques (PALM, spt-PALM, or FLIM). Its photostability large enough obtain fluorescence, surface enhanced...

Abstract Single-molecule imaging promises the observation of individual molecules at work in living cells 1,2 . In plants, however, tracking single is generally limited to mere hundred milliseconds 3–5 , making it virtually impossible observe live dynamic cellular events with molecular resolution. Here, we introduce photochromic reversion which uses EOS fluorescent protein’s dark state upon blue light illumination 6 thereby stabilizing and extending single-molecule particle photoactivated...

Malignant primary brain tumors are a group of highly aggressive and often infiltrating that lack adequate therapeutic treatments to achieve long time survival. Complete tumor removal is one precondition reach this goal. A promising approach optimize resection margins eliminate remaining infiltrative so-called guerilla cells photodynamic therapy (PDT) using organic photosensitizers can pass the disrupted blood-brain-barrier selectively accumulate in tissue. Hypericin fulfills these conditions...

Abstract The first step in photosynthesis is an extremely efficient energy transfer mechanism that led to the debate which extent quantum coherence may be involved between photosynthetic pigments. In search of such a coherent behavior, we have embedded living cyanobacteria parallel mirrors optical microresonator irradiated with low intensity white light. As consequence, observe vacuum Rabi splitting transmission and fluorescence spectra as result strong light matter coupling chlorophyll...

Photosynthesis is one the most important biological processes on earth, producing life-giving oxygen, and basis for a large variety of plant products. Measurable properties photosynthesis provide information about its biophysical state, in turn, physiological conditions photoautotrophic organism. For instance, chlorophyll fluorescence intensity an intact photosystem not constant as case single fluorescent dye solution but shows temporal changes related to quantum yield photosystem....

Abstract Protein-protein interaction studies provide valuable insights into cellular signaling. The well-studied brassinosteroid (BR) signaling is initiated by the hormone-binding receptor Brassinosteroid Insensitive 1 (BRI1) and its co-receptor BRI1 Associated Kinase (BAK1). BAK1 were shown to interact independently with Receptor-Like Protein 44 (RLP44), which implicated in BRI1/BAK1-dependent cell wall integrity perception. To demonstrate proposed complex formation of BRI1, RLP44, we...

ABSTRACT Super-resolution microscopy (SRM) approaches revolutionize cell biology by providing insights into the nanoscale organization and dynamics of macromolecular assemblies single molecules in living cells. A major hurdle limiting SRM democratization is post-acquisition data analysis which often complex time-consuming. Here, we present OneFlowTraX, a user-friendly open-source software dedicated to single-molecule localization (SMLM) such as single-particle tracking photoactivated...

ABSTRACT Super-resolution microscopy techniques have revolutionized cell biology by providing insights into the dynamics of single molecules and nanoscale organization within living cells. However, application dynamic live-cell methods in plants has been limited lack suitable fluorophores for simultaneous visualization multiple proteins. To address this challenge, we implemented a two-color sptPALM approach using codon-optimized photoactivatable fluorescent proteins PA-GFP PATagRFP....

Abstract Traditional models such as the fluid mosaic model or lipid raft hypothesis have shaped our understanding of plasma membrane (PM) organization. However, recent discoveries extended these paradigms by pointing to existence micro- and nanodomains. Here, we investigated role cytoskeleton in general whether picket fence model, established animal cells, is transferable plant cell system. By using single-particle tracking photoactivated localization microscopy (sptPALM) combination with...

Malignant primary brain tumors are a group of highly aggressive and often infiltrating that lack adequate therapeutic treatments to achieve long time survival. Complete tumor removal is one precondition reach this goal. A promising approach optimize resection margins eliminate remaining infiltrative so-called guerilla cells photodynamic therapy (PDT) using organic photosensitizers can pass the disrupted blood-brain-barrier selectively accumulate in tissue. Hypericin fulfills these conditions...

The yellow fluorescent protein (YFP) is frequently used in a complementation assay called bimolecular fluorescence (BiFC), and employed to visualize protein–protein interactions. In this analysis, two different, nonfluorescent fragments of YFP are genetically attached proteins interest. Upon interaction these proteins, the brought into proximity close enough reconstitute their original structure, enabling fluorescence. BiFC allows for straightforward readout interactions furthermore...

Abstract As technical developments in omics and biomedical imaging drive the increase quality, modality, throughput of data generation life sciences, need for information systems capable integrative, long-term storage management these heterogeneous digital assets is also increasing. Here, we propose an approach based on principles Service Oriented Architecture design, to allow integrated analysis multi-omics data. The proposed architecture introduces interoperable image system, OMERO server,...

Abstract The first step in photosynthesis is an extremely efficient energy transfer mechanism, which difficult to be explained by classical short-range migration (“hopping”) and led the debate extent quantum coherence involved between photosynthetic pigments. Embedding living cyanobacteria mirrors of optical microresonator using low intensity white light irradiation we observe vacuum Rabi splitting transmission fluorescence spectra as a result strong matter coupling chlorophyll resonator...

Photosynthesis is one the most important biological processes on earth, producing life-giving oxygen and basis for a large variety of plant products. Measurable properties photosynthesis provide information about its biophysical state and, in turn, physiological conditions photoautotrophic organism. For instance, chlorophyll fluorescence an intact photosystem not linear as case single fluorescent dye solution, but shows temporal changes related to quantum yield photosystem. Commercial...