2000
DOI: 10.1105/tpc.12.11.2219
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In Situ Localization and in Vitro Induction of Plant COPI-Coated Vesicles

Abstract: Coat protein (COP)-coated vesicles have been shown to mediate protein transport through early steps of the secretory pathway in yeast and mammalian cells. Here, we attempt to elucidate their role in vesicular trafficking of plant cells, using a combined biochemical and ultrastructural approach. Immunogold labeling of cryosections revealed that COPI proteins are localized to microvesicles surrounding or budding from the Golgi apparatus. COPI-coated buds primarily reside on the cis -face of the Golgi stack. In a… Show more

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Cited by 181 publications
(164 citation statements)
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“…To examine whether the ST:GFP-positive punctate stains represented the Golgi apparatus, the localization of ST:GFP was compared with that of endogenous g-COP, a COPI component localized to the Golgi apparatus (Pimpl et al, 2000). The localization of ST:GFP and g-COP was determined directly by GFP fluorescence and immunohistochemistry using anti-g-COP antibody (Pimpl et al, 2000), respectively. In both the presence and absence of HA:AtPRA1.B6, the endogenous g-COP and ST:GFP produced punctate staining patterns (Fig.…”
Section: Ha:atpra1b6 Does Not Inhibit the Trafficking Of Proteins Lomentioning
confidence: 99%
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“…To examine whether the ST:GFP-positive punctate stains represented the Golgi apparatus, the localization of ST:GFP was compared with that of endogenous g-COP, a COPI component localized to the Golgi apparatus (Pimpl et al, 2000). The localization of ST:GFP and g-COP was determined directly by GFP fluorescence and immunohistochemistry using anti-g-COP antibody (Pimpl et al, 2000), respectively. In both the presence and absence of HA:AtPRA1.B6, the endogenous g-COP and ST:GFP produced punctate staining patterns (Fig.…”
Section: Ha:atpra1b6 Does Not Inhibit the Trafficking Of Proteins Lomentioning
confidence: 99%
“…For immunostaining, protoplasts on coverslips were fixed with 4% (v/v) paraformaldehyde, as described previously (Frigerio et al, 1998;Park et al, 2004). Fixed protoplasts were labeled with anti-g-COP antibody (Pimpl et al, 2000). Cells were washed with buffer (10 mM Tris, pH 7.4, 0.9% [w/v] NaCl, 0.25% [w/v] gelatin, 0.02% [w/v] SDS, and 0.1% [v/v] Triton X-100), and secondary immunostaining was performed for 3 h using tetramethylrhodamine-5-(and-6)-isothiocyanate-labeled goat anti-rabbit antibodies (Molecular Probes).…”
Section: Transient Expression Immunofluorescence Staining and Micromentioning
confidence: 99%
“…Unlike Arabidopsis (Arabidopsis thaliana) root cells where ARF1 localizes to both endosomes and the Golgi apparatus (Xu and Scheres, 2005), in tobacco BY-2 cells ARF1 is restricted to the rims of Golgi cisternae, where COPI vesicles are formed (Pimpl et al, 2000;Ritzenthaler et al, 2002;Yang et al, 2005;Robinson and Ritzenthaler, 2006). We have therefore used ARF1 immunofluorescence as a way of monitoring the reconstitution of BY-2 Golgi stacks during BFA washout.…”
Section: Arf1 Immunolabeling Of By-2 Golgi Stacks Regenerating After mentioning
confidence: 99%
“…OsSRT1 (ref. 45) and Osb 0 -COP 46 were selected as the nuclear and cytoplasmic protein controls. Anti-TMS5, anti-OsSRT1 and anti-Osb 0 -COP were generated by immunizing rabbits.…”
Section: Subcellular Localizationmentioning
confidence: 99%