Publications by authors named "Masayoshi Nakamura"

Article Synopsis
  • A water-soluble aromatic nanobelt was created through a straightforward reaction that modified a parent compound, resulting in an alkyne-functionalized version.
  • This modified nanobelt underwent a specific chemical reaction to attach a dye, making it suitable for cellular studies.
  • Experiments with HeLa cells showed unique uptake patterns of the nanobelt, which were influenced by its belt-like shape, as confirmed by control experiments and theoretical analysis.
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Plant cell cortical microtubules are located beneath the plasma membrane and direct the location of cellulose synthases during interphase, influencing cell morphology. Microtubule-associated proteins (MAPs) regulate these microtubules in response to growth and environmental stimuli. This review focuses on recent advances in understanding microtubule nucleation mechanisms in plants and the spatiotemporal regulation of cortical arrays via phytohormone signaling.

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  • Researchers discovered that plant cell nuclei emit natural fluorescence in the near-infrared (NIR) spectrum, allowing for a non-invasive imaging technique in live cells.
  • This NIR imaging method was successfully used to observe the dynamic behavior of nuclei in key plant structures like roots and pollen tubes, where other organelles have limited autofluorescence.
  • The technique, driven by the phytochrome protein, was shown to work across various plant species and does not require genetic modifications, making it a valuable tool for studying both model and non-model plants.
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  • Microtubule severing by katanin is crucial for the formation and patterning of dynamic microtubule arrays in response to plant growth and environmental changes.
  • Dysfunction in katanin activity can lead to growth and division issues in plant cells.
  • Katanin is strategically recruited to specific sites on microtubules, particularly at intersections and nucleation sites, which is vital for proper microtubule maintenance and organization, especially during cell division.
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SWEET sucrose transporters play important roles in the allocation of sucrose in plants. Some SWEETs were shown to also mediate transport of the plant growth regulator gibberellin (GA). The close physiological relationship between sucrose and GA raised the questions of whether there is a functional connection and whether one or both of the substrates are physiologically relevant.

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  • Understanding cellular processes in multicellular organisms requires knowledge of small molecules, enzyme activities, and protein functions within cells and tissues.
  • The distribution of receptors, ligands, and metabolites must be integrated to gain insights into metabolic and signaling dynamics for in vivo biochemistry.
  • Advancements in genetically encoded fluorescent sensors and imaging technologies have made it possible to monitor cellular activities and metabolite distribution in live plants, presenting both opportunities and challenges in sensor design and application.
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SWEETs play important roles in intercellular sugar transport. Induction of SWEET sugar transporters by Transcription Activator-Like effectors (TALe) of Xanthomonas ssp. is key for virulence in rice, cassava and cotton.

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  • This paper examines three perspectives on safety management in Japan post-COVID-19.
  • First, it discusses how society has shifted from aiming for zero risk to accepting some level of risk to sustain social activities.
  • Second, it highlights a transformation in work practices, moving towards a hybrid model of remote and in-person work, with a focus on job evaluation over seniority.
  • Lastly, it critiques the limitations of Japan's group-oriented societal systems and emphasizes the need for systemic changes to address issues like vaccine development and healthcare challenges.
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Fluorescent probes are powerful tools for visualizing cellular and subcellular structures, their dynamics and cellular molecules in living cells and enable us to monitor cellular processes in a spatiotemporal manner within complex and crowded systems. In addition to popular fluorescent proteins, a wide variety of small-molecule dyes have been synthesized through close association with the interdisciplinary field of chemistry and biology, ranging from those suitable for labeling cellular compartments such as organelles to those for labeling intracellular biochemical and biophysical processes and signaling. In recent years, self-labeling technologies including the SNAP-tag system have allowed us to attach these dyes to cellular domains or specific proteins and are beginning to be employed in plant studies.

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Microtubules are severed by katanin at distinct cellular locations to facilitate reorientation or amplification of dynamic microtubule arrays, but katanin targeting mechanisms are poorly understood. Here we show that a centrosomal microtubule-anchoring complex is used to recruit katanin in acentrosomal plant cells. The conserved protein complex of Msd1 (also known as SSX2IP) and Wdr8 is localized at microtubule nucleation sites along the microtubule lattice in interphase Arabidopsis cells.

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Plant hormones play important roles in plant growth and development and physiology, and in acclimation to environmental changes. The hormone signaling networks are highly complex and interconnected. It is thus important to not only know where the hormones are produced, how they are transported and how and where they are perceived, but also to monitor their distribution quantitatively, ideally in a non-invasive manner.

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  • Fluorescent biosensors, especially those based on fluorescent proteins (FPs), are effective for monitoring cellular processes in live organisms due to their selectivity and minimal invasiveness.
  • They have been widely used in plant research to track changes in various factors like pH, ion concentration, and redox state.
  • The chapter highlights the application of FP-based biosensors in plants, particularly focusing on monitoring intracellular calcium dynamics in Arabidopsis thaliana using a specific genetically encoded Ca indicator.
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  • Synthetic chemical fluorescent dyes offer promising applications in biology for studying specific proteins and cellular processes through targeted labeling techniques like SNAP-tag.
  • Of 31 tested synthetic dyes, 23 were successfully absorbed by BY-2 plant cells, indicating their potential in measuring endocytosis.
  • Effective labeling of proteins, such as α-tubulin and PIN-FORMED2, was demonstrated using selected dyes, enabling detailed observation of cellular dynamics in Arabidopsis seedlings.
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Background: Sugar content is an important determinant of fruit sweetness, but details on the complex molecular mechanism underlying fruit sugar accumulation remain scarce. Here, we report the role of sucrose transporter (SUT) family in regulating fruit sugar accumulation in apple.

Results: Gene-tagged markers were developed to conduct candidate gene-based association study, and an SUT4 member MdSUT4.

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Cytokinesis is fundamental for cell proliferation [1, 2]. In plants, a bipolar short-microtubule array forms the phragmoplast, which mediates vesicle transport to the midzone and guides the formation of cell walls that separate the mother cell into two daughter cells [2]. The phragmoplast centrifugally expands toward the cell cortex to guide cell-plate formation at the cortical division site [3, 4].

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  • Nucleation has been the main focus of microtubule generation studies, but severing is highlighted as an alternative mechanism that creates new polymer ends in response to light in plants.
  • CLASP is identified as a key protein that stabilizes new plus ends formed by severing, which is crucial for the correct reorientation of microtubule arrays in plant cells.
  • Mutants lacking CLASP show impaired stabilization of these ends and reduced microtubule amplification, revealing the importance of CLASP in this process through computational modeling.
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The cortical microtubule arrays of higher plants are organized without centrosomes and feature treadmilling polymers that are dynamic at both ends. The control of polymer end stability is fundamental for the assembly and organization of cytoskeletal arrays, yet relatively little is understood about how microtubule minus ends are controlled in acentrosomal microtubule arrays, and no factors have been identified that act at the treadmilling minus ends in higher plants. Here, we identify SPIRAL2 (SPR2) as a protein that tracks minus ends and protects them against subunit loss.

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Article Synopsis
  • In higher plants, cortical microtubule reorientation is crucial for adapting cell growth to environmental changes, though the exact process is not well understood.
  • Recent studies using genetics and live cell imaging have revealed mechanisms that regulate microtubule nucleation and severing necessary for their reorientation, especially in response to blue light.
  • KATANIN plays a key role by releasing microtubules from their nucleation sites and enhancing the formation of new microtubules through severing, allowing for efficient transitions in microtubule arrays.
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Article Synopsis
  • Many differentiated animal cells and all higher plant cells form microtubule structures during interphase without a central organizer like a centrosome, leading to essential functions such as morphogenesis.
  • In higher plants, microtubule nucleation occurs through distributed complexes at the cell cortex, which can create new microtubules either parallel to existing ones or at about a 40° branching angle.
  • The protein GCP-WD plays a crucial role in positioning γ-tubulin ring complexes for nucleation and enhances branching formation in microtubule arrays, allowing plant cells to create various patterns and structures.
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Environmental and hormonal signals cause reorganization of microtubule arrays in higher plants, but the mechanisms driving these transitions have remained elusive. The organization of these arrays is required to direct morphogenesis. We discovered that microtubule severing by the protein katanin plays a crucial and unexpected role in the reorientation of cortical arrays, as triggered by blue light.

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Microtubules in eukaryotic cells are nucleated from ring-shaped complexes that contain γ-tubulin and a family of homologous γ-tubulin complex proteins (GCPs), but the subunit composition of the complexes can vary among fungi, animals and plants. Arabidopsis GCP3-interacting protein 1 (GIP1), a small protein with no homology to the GCP family, interacts with GCP3 in vitro, and is a plant homolog of vertebrate mitotic-spindle organizing protein associated with a ring of γ-tubulin 1 (MOZART1), a recently identified component of the γ-tubulin complex in human cell lines. In this study, we characterized two closely related Arabidopsis GIP1s: GIP1a and GIP1b.

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  • Organelles like RNA processing bodies, Golgi bodies, peroxisomes, and mitochondria rely on actin filaments for transport but pause at cortical microtubules during their movement.
  • The study shows that removing microtubules does not significantly impact how often these organelles pause, suggesting that pausing is a common feature of their motility.
  • The research indicates that while microtubules help form stable junctions with the endoplasmic reticulum (ER), they are not necessary for the pausing process, potentially aiding interactions between the ER and moving organelles.
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Pyruvate serves as a metabolic precursor for many plastid-localized biosynthetic pathways, such as those for fatty acids, terpenoids and branched-chain amino acids. In spite of the importance of pyruvate uptake into plastids (organelles within cells of plants and algae), the molecular mechanisms of this uptake have not yet been explored. This is mainly because pyruvate is a relatively small compound that is able to passively permeate lipid bilayers, which precludes accurate measurement of pyruvate transport activity in reconstituted liposomes.

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Microtubule nucleation in interphase plant cells primarily occurs through branching from pre-existing microtubules at dispersed sites in the cell cortex. The minus ends of new microtubules are often released from the sites of nucleation, and the free microtubules are then transported to new locations by polymer treadmilling. These nucleation-and-release events are characteristic features of plant arrays in interphase cells, but little is known about the spatiotemporal control of these events by nucleating protein complexes.

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