bioRxiv Science⌕ Search

Biology subjects

Torii, K.

Publications and source records attributed to Torii, K..

4 recordsLinked to original sources

Near-Infrared Turn-On Fluorogenic Probe for Versatile Detection of Inorganic Polyphosphates

Inorganic polyphosphate (polyP) is a ubiquitous phosphate biopolymer involved in diverse cellular processes. Despite its significance, selective detection of polyP remains challenging because of its simple and highly charged structure. Here, we report a near-infrared (NIR) fluorogenic turn-on chemosensor for selective polyP detection and imaging, SiX-DPA-Zn. The probe combines a silicon-xanthene (SiX) fluorophore with a zinc(II)-coordinated 2,2'-dipicolylamine (DPA-Zn2+) recognition unit and shows more than 100-fold selectivity for inorganic polyP over ADP and ATP. SiX-DPA-Zn enables quantitative detection of polyP at micromolar concentrations in microplate assays and stains a broad range of polyP species, starting from tripolyphosphate, in polyacrylamide gels. In HEK293 cells expressing Escherichia coli polyphosphate kinase 1, the probe visualizes intracellular polyP and enables quantitative analysis of polyP levels in relation to nuclear proteins for example fibrillarin and nucleolin. Stimulated emission depletion (STED) microscopy further revealed subdiffraction-sized polyP granules within polyP aggregates. SiX-DPA-Zn is the first near-infrared (NIR) fluorogenic chemosensor for polyP that is compatible with multiple detection platforms, including microplate assays, polyacrylamide gel staining, confocal and super-resolution STED microscopy.

biochemistry↗

Nature and effective range of non-cell autonomous activator and inhibitor peptides specifying plant stomatal patterning

Stomata are epidermal valves that facilitate gas exchange between plants and their environment. Stomatal patterning is regulated by EPIDERMAL PATTERING FACTOR (EPF)-family of secreted peptides: EPF1 enforcing stomatal spacing, whereas EPF-LIKE9, also known as Stomagen, promoting stomatal development. It remains unknown, however, how far these signaling peptides act. Utilizing Cre-Lox recombination-based mosaic sectors that overexpress either EPF1 or Stomagen in Arabidopsis cotyledons, we reveal a range within the epidermis and across the cell layers in which these peptides influence patterns. To quantitatively determine their effective ranges, we developed a computational pipeline, SPACE (Stomata Patterning AutoCorrelation on Epidermis), that describes probabilistic two-dimensional stomatal distributions based upon spatial autocorrelation statistics used in Astrophysics. The SPACE analysis shows that, whereas both peptides act locally, the inhibitor, EPF1, exerts longer-range effects than the activator, Stomagen. Furthermore, local perturbation of stomatal development has little influence on global two-dimensional stomatal patterning. Our findings conclusively demonstrate the nature and extent of EPF peptides as non-cell autonomous local signals and provides a means to quantitatively characterize complex spatial patterns in development. Summary StatementNon-cell autonomous effects of activator and inhibitor peptides on 2-D spatial patterning of stomata were quantitatively characterized using chimeric sectors and a SPACE computational pipeline.

plant biology↗

Origination of the circadian clock system in stem cells regulates cell differentiation in Arabidopsis thaliana.

The circadian clock regulates various physiological responses. To achieve this, both animals and plants have distinct circadian clocks in each tissue that are optimized for that tissues respective functions. However, if and how the tissue-specific circadian clocks are involved in specification of cell types remains unclear. Here, by implementing a single-cell transcriptome with a new analytics pipeline, we have reconstructed an actual time-series of the cell differentiation process at single-cell resolution, and discovered that the Arabidopsis circadian clock is involved in the process of cell differentiation through transcription factor BRI1-EMS SUPPRESSOR 1 (BES1) signaling. In this pathway, direct repression of LATE ELONGATED HYPOCOTYL (LHY) expression by BES1 triggers reconstruction of the circadian clock in stem cells. The reconstructed circadian clock regulates cell differentiation through fine-tuning of key factors for epigenetic modification, cell-fate determination, and the cell cycle. Thus, the establishment of circadian systems precedes cell differentiation and specifies cell types.

plant biology↗

MUTE Directly Orchestrates Cell State Switch and the Single Symmetric Division to Create Stomata

Precise cell division control is critical for developmental patterning. For the differentiation of a functional stoma, a cellular valve for efficient gas exchange, the single symmetric division of an immediate precursor is absolutely essential. Yet, the mechanism governing the single division event remains unclear. Here we report the complete inventories of gene expression by the Arabidopsis bHLH protein MUTE, a potent inducer of stomatal differentiation. MUTE switches the gene expression program initiated by its sister bHLH, SPEECHLESS. MUTE directly induces a suite of cell-cycle genes, including CYCD5;1, and their transcriptional repressors, FAMA and FOUR LIPS. The architecture of the regulatory network initiated by MUTE represents an Incoherent Type 1 Feed-Forward Loop. Our mathematical modeling and experimental perturbations support a notion that MUTE orchestrates a transcriptional cascade leading to the tightly-restricted, robust pulse of cell-cycle gene expression, thereby ensuring the single cell division to create functional stomata.\n\nHighlightsO_LIComplete inventories of gene expression in stomatal differentiation state are elucidated\nC_LIO_LIMUTE switches stomatal patterning program initiated by its sister bHLH, SPEECHLESS\nC_LIO_LIMUTE directly induces cell-cycle genes and their direct transcriptional repressors\nC_LIO_LIIncoherent feed-forward loop by MUTE ensures the single division of a stomatal precursor\nC_LI

plant biology↗