Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure

Watanabe, Tomonobu M.; Imada, Katsumi; Yoshizawa, Keiko; Nishiyama, Masayoshi; Kato, Chiaki; Abe, Fumiyoshi; Morikawa, Takamitsu; Kinoshita, Miki; Fujita, Hideaki; Yanagida, Toshio

doi:10.1371/journal.pone.0073212

Cited by 24 publications

(29 citation statements)

References 35 publications

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“…Fine-tip pressure probing or nano-indentation has the potential to deliver such a measurement strategy. In parallel to the mechanical measurement, an optical approach should also be employed; pressure-sensitive proteins, including fluorescent markers, can be engineered as biosensors of turgor pressure ( Barstow et al ., 2008 , 2009 ; Watanabe et al ., 2013 ).…”

Section: Discussionmentioning

confidence: 99%

Flowers under pressure: ins and outs of turgor regulation in development

2014

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BackgroundTurgor pressure is an essential feature of plants; however, whereas its physiological importance is unequivocally recognized, its relevance to development is often reduced to a role in cell elongation.ScopeThis review surveys the roles of turgor in development, the molecular mechanisms of turgor regulation and the methods used to measure turgor and related quantities, while also covering the basic concepts associated with water potential and water flow in plants. Three key processes in flower development are then considered more specifically: flower opening, anther dehiscence and pollen tube growth.ConclusionsMany molecular determinants of turgor and its regulation have been characterized, while a number of methods are now available to quantify water potential, turgor and hydraulic conductivity. Data on flower opening, anther dehiscence and lateral root emergence suggest that turgor needs to be finely tuned during development, both spatially and temporally. It is anticipated that a combination of biological experiments and physical measurements will reinforce the existing data and reveal unexpected roles of turgor in development.

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Section: Discussionmentioning

confidence: 99%

Flowers under pressure: ins and outs of turgor regulation in development

2014

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“…In cell culture, FRET has also been used to measure uniaxial compression (Paszek et al, 2014). More generally, fluorescent materials exhibit a variety of properties that could be dependent on other mechanical quantities, such as intracellular pressure (Gomez-Martinez et al, 2013;Watanabe et al, 2013), and thus could be used to develop new force probes.…”

Section: Fretmentioning

confidence: 99%

Measuring forces and stressesin situin living tissues

2016

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Development, homeostasis and regeneration of tissues result from a complex combination of genetics and mechanics, and progresses in the former have been quicker than in the latter. Measurements of in situ forces and stresses appear to be increasingly important to delineate the role of mechanics in development. We review here several emerging techniques: contact manipulation, manipulation using light, visual sensors, and non-mechanical observation techniques. We compare their fields of applications, their advantages and limitations, and their validations. These techniques complement measurements of deformations and of mechanical properties. We argue that such approaches could have a significant impact on our understanding of the development of living tissues in the near future.

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“…GimRET was first developed by fusing the cyan fluorescent protein (CFP) with a glycine-inserted yellow fluorescent protein (YFP1G) 18 . The fluorescence intensity of YFP1G decreases as the surrounding concentration of protein increases (Figure S1) 16 .…”

Section: Resultsmentioning

confidence: 99%

In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis

Konishi

Asai

Watanabe³

et al. 2017

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The central channel of the nuclear pore complex (NPC) is occupied by non-structured polypeptides with a high content of Phe-Gly (FG) motifs. This protein-rich environment functions as an entropic barrier that prevents the passage of molecules, as well as the binding sites for karyopherins, to regulate macromolecular traffic between the nucleoplasm and the cytoplasm. In this study, we expressed individual Nups fused with a crowding-sensitive probe (GimRET) to determine the spatial distribution of protein-rich domains within the central channel in vivo, and characterize the properties of the entropic barrier. Analyses of the probe signal revealed that the central channel contains two protein-rich domains at both the nucleoplasmic and cytoplasmic peripheries, and a less-crowded central cavity. Karyopherins and other soluble proteins are not the constituents of the protein-rich domains. The time-lapse observation of the post-mitotic reassembly process also revealed how individual protein-rich domains are constructed by a sequential assembly of nucleoporins.

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Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure

Cited by 24 publications

References 35 publications

Flowers under pressure: ins and outs of turgor regulation in development

Flowers under pressure: ins and outs of turgor regulation in development

Measuring forces and stressesin situin living tissues

In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis

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