• Molecules and Cells
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• v.45 no.1
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• pp.6-15
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• 2022

Phase separation is a thermodynamic process leading to the formation of compositionally distinct phases. For the past few years, numerous works have shown that biomolecular phase separation serves as biogenesis mechanisms of diverse intracellular condensates, and aberrant phase transitions are associated with disease states such as neurodegenerative diseases and cancers. Condensates exhibit rich phase behaviors including multiphase internal structuring, noise buffering, and compositional tunability. Recent studies have begun to uncover how a network of intermolecular interactions can give rise to various biophysical features of condensates. Here, we review phase behaviors of biomolecules, particularly with regard to regular solution models of binary and ternary mixtures. We discuss how these theoretical frameworks explain many aspects of the assembly, composition, and miscibility of diverse biomolecular phases, and highlight how a model-based approach can help elucidate the detailed thermodynamic principle for multicomponent intracellular phase separation.

• Mycobiology
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• v.51 no.5
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• pp.273-280
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• 2023

The nucleolus is the largest, membrane-less organelle within the nucleus of eukaryotic cell that plays a critical role in rRNA transcription and assembly of ribosomes. Recently, the nucleolus has been shown to be implicated in an array of processes including the formation of signal recognition particles and response to cellular stress. Such diverse functions of nucleolus are mediated by nucleolar proteins. In this study, we characterized a gene coding a putative protein containing a nucleolar localization sequence (NoLS) in the rice blast fungus, Magnaporthe oryzae. Phylogenetic and domain analysis suggested that the protein is orthologous to Rrp8 in Saccharomyces cerevisiae. MoRRP8-GFP (translational fusion of MoRRP8 with green fluorescence protein) co-localizes with a nucleolar marker protein, MoNOP1 fused to red fluorescence protein (RFP), indicating that MoRRP8 is a nucleolar protein. Deletion of the MoRRP8 gene caused a reduction in vegetative growth and impinged largely on asexual sporulation. Although the asexual spores of DMorrp8 were morphologically indistinguishable from those of wild-type, they showed delay in germination and reduction in appressorium formation. Our pathogenicity assay revealed that the MoRRP8 is required for full virulence and growth within host plants. Taken together, these results suggest that nucleolar processes mediated by MoRRP8 is pivotal for fungal development and pathogenesis.

• Applied Microscopy
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• v.39 no.4
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• pp.333-340
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• 2009

In the present study, ultrastructural features of the mesophyll tissue have been investigated in Crassulacean acid metabolism (CAM)-performing succulent Orostachys. A large central vacuole and numerous small vacuoles in the peripheral cytoplasm were characterized at the subcellular level in both developing and mature mesophyll cells. The most notable feature was the invagination of vacuolar membranes into the secondary vacuoles or multivesicular bodies. In many cases, tens of single, membrane-bound secondary vacuoles of various sizes were found to be formed within the central vacuole. multivesicular bodies containing numerous small vesicles were also distributed in the cytoplasm but were better developed within the central vacuole. Occasionally, electron-dense prevacuolar compartments, directly attached to structures appearing to be small vacuoles, were also detected in the cytoplasm. One or more huge central vacuoles were frequently observed in cells undergoing differentiation and maturation. Consistent with the known occurrence of morphologically distinct vacuoles within different tissues, two types of vacuoles, one representing lytic vacuoles and the other, most likely protein storage vacuoles, were noted frequently within Orostachys mesophyll. The two types coexisted in mature vegetative cells but did not merge during the study. Nevertheless, the coexistence of two distinct vacuole types in maturing cells implies the presence of more than one mechanism for vacuolar solute sorting in these species. The vacuolar membrane is known to be unique among the intracellular compartments for having different channels and/or pumps to maintain its function. In CAM plants, the vacuole is a very important organelle that regulates malic acid diurnal fluctuation to a large extent. The membrane invagination seen in Orostachys mesophyll likely plays a significant role in survival under the physiological drought conditions in which these Orostachys occur; by increasing to such a large vacuolar volume, the mesophyll cells are able to retain enormous amounts of acid when needed. Furthermore, the mesophyll cells are able to attain their large sizes with less energy expenditure in order to regulate the large degree of diurnal fluctuation of organic acid that occurs within the vacuoles of Orostachys.