Proliferation of the Golgi apparatus in tobacco BY-2 cells during cell proliferation after release from the stationary phase of growth

Abiodun, Moses Olabiyi; Matsuoka, Ken

doi:10.4161/psb.25027

Cited by 3 publications

(2 citation statements)

References 11 publications

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“…Our observations here and our recent analysis of Golgi proliferation using a monomeric KikGR-tagged reporter protein (Abiodun and Matsuoka, 2013a , b ) indicate that bulk conversion of the reporter fluorescence color and subsequent analysis is a method that can be applied to analyze the synthesis and/or turnover of distinct intracellular structures. However, this method has a limitation when used for the quantitative comparison of preexisting and newly synthesized proteins.…”

Section: Discussionsupporting

confidence: 52%

“…We have reported that fluorescence of a Golgi-targeted fusion protein comprising of a prolyl-hydroxylase NtP4H1.1 and monomeric KikGR could be converted under illumination by purple light, and this conversion allowed us to monitor the proliferation of the Golgi apparatus in tobacco BY-2 cells (Abiodun and Matsuoka, 2013a , b ). We used the same conversion apparatus (Abiodun and Matsuoka, 2013a ) to convert the fluorescence of the color of the aggregate from green to red.…”

Section: Resultsmentioning

confidence: 99%

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Monitoring protein turnover during phosphate starvation-dependent autophagic degradation using a photoconvertible fluorescent protein aggregate in tobacco BY-2 cells

2014

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We have developed a system for quantitative monitoring of autophagic degradation in transformed tobacco BY-2 cells using an aggregate-prone protein comprised of cytochrome b5 (Cyt b5) and a tetrameric red fluorescent protein (RFP). Unfortunately, this system is of limited use for monitoring the kinetics of autophagic degradation because the proteins synthesized before and after induction of autophagy cannot be distinguished. To overcome this problem, we developed a system using kikume green-red (KikGR), a photoconvertible and tetrameric fluorescent protein that changes its fluorescence from green to red upon irradiation with purple light. Using the fusion protein of Cyt b5 and KikGR together with a method for the bulk conversion of KikGR, which we had previously used to convert the Golgi-localized monomeric KikGR fusion protein, we were able to monitor both the growth and de novo formation of aggregates. Using this system, we found that tobacco cells do not cease protein synthesis under conditions of phosphate (Pi)-starvation. Induction of autophagy under Pi-starvation, but not under sugar- or nitrogen-starvation, was specifically inhibited by phosphite, which is an analog of Pi with a different oxidation number. Therefore, the mechanism by which BY-2 cells can sense Pi-starvation and induce autophagy does not involve sensing a general decrease in energy supply and a specific Pi sensor might be involved in the induction of autophagy under Pi-starvation.

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Section: Discussionsupporting

confidence: 52%

Section: Resultsmentioning

confidence: 99%

Monitoring protein turnover during phosphate starvation-dependent autophagic degradation using a photoconvertible fluorescent protein aggregate in tobacco BY-2 cells

2014

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Using Photoconvertible and Extractable Fluorescent Proteins to Study Autophagy in Plants

Abiodun

Matsuoka

2017

Methods in Enzymology

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Wide-Range High-Resolution Transmission Electron Microscopy Reveals Morphological and Distributional Changes of Endomembrane Compartments during Log to Stationary Transition of Growth Phase in Tobacco BY-2 Cells

Toyooka

Sato

Kutsuna

et al. 2014

Plant and Cell Physiology

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Rapid growth of plant cells by cell division and expansion requires an endomembrane trafficking system. The endomembrane compartments, such as the Golgi stacks, endosome and vesicles, are important in the synthesis and trafficking of cell wall materials during cell elongation. However, changes in the morphology, distribution and number of these compartments during the different stages of cell proliferation and differentiation have not yet been clarified. In this study, we examined these changes at the ultrastructural level in tobacco Bright yellow 2 (BY-2) cells during the log and stationary phases of growth. We analyzed images of the BY-2 cells prepared by the high-pressure freezing/freeze substitution technique with the aid of an auto-acquisition transmission electron microscope system. We quantified the distribution of secretory and endosomal compartments in longitudinal sections of whole cells by using wide-range gigapixel-class images obtained by merging thousands of transmission electron micrographs. During the log phase, all Golgi stacks were composed of several thick cisternae. Approximately 20 vesicle clusters (VCs), including the trans-Golgi network and secretory vesicle cluster, were observed throughout the cell. In the stationary-phase cells, Golgi stacks were thin with small cisternae, and only a few VCs were observed. Nearly the same number of multivesicular body and small high-density vesicles were observed in both the stationary and log phases. Results from electron microscopy and live fluorescence imaging indicate that the morphology and distribution of secretory-related compartments dramatically change when cells transition from log to stationary phases of growth.

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Proliferation of the Golgi apparatus in tobacco BY-2 cells during cell proliferation after release from the stationary phase of growth

Cited by 3 publications

References 11 publications

Monitoring protein turnover during phosphate starvation-dependent autophagic degradation using a photoconvertible fluorescent protein aggregate in tobacco BY-2 cells

Monitoring protein turnover during phosphate starvation-dependent autophagic degradation using a photoconvertible fluorescent protein aggregate in tobacco BY-2 cells

Using Photoconvertible and Extractable Fluorescent Proteins to Study Autophagy in Plants

Wide-Range High-Resolution Transmission Electron Microscopy Reveals Morphological and Distributional Changes of Endomembrane Compartments during Log to Stationary Transition of Growth Phase in Tobacco BY-2 Cells

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