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dc.contributor.author
Vermeer, Joop E.M.
dc.contributor.author
Van Leeuwen, Wessel
dc.contributor.author
Tobeña Santamaria, Rafa
dc.contributor.author
Laxalt, Ana Maria
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dc.contributor.author
Jones, David R.
dc.contributor.author
Divecha, Nullin
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Gadella Jr., Theodorus W.J.
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Munnik, Teun
dc.date.available
2023-01-06T00:32:08Z
dc.date.issued
2006-12
dc.identifier.citation
Vermeer, Joop E.M.; Van Leeuwen, Wessel; Tobeña Santamaria, Rafa; Laxalt, Ana Maria; Jones, David R.; et al.; Visualization of PtdIns3P dynamics in living plant cells; Wiley Blackwell Publishing, Inc; Plant Journal; 47; 5; 12-2006; 687-700
dc.identifier.issn
0960-7412
dc.identifier.uri
http://hdl.handle.net/11336/183653
dc.description.abstract
To investigate PtdIns3P localization and function in plants, a fluorescent PtdIns3P-specific biosensor (YFP-2xFYVE) was created. On lipid dot blots it bound specifically and with high affinity to PtdIns3P. Transient expression in cowpea protoplasts labelled vacuolar membranes and highly motile structures undergoing fusion and fission. Stable expression in tobacco BY-2 cells labelled similar motile structures, but labelled vacuolar membranes hardly at all. YFP-2xFYVE fluorescence strongly co-localized with the pre-vacuolar marker AtRABF2b, partially co-localized with the endosomal tracer FM4-64, but showed no overlap with the Golgi marker STtmd-CFP. Treatment of cells with wortmannin, a PI3 kinase inhibitor, caused the YFP-2xFYVE fluorescence to redistribute into the cytosol and nucleus within 15 min. BY-2 cells expressing YFP-2xFYVE contained twice as much PtdIns3P as YFP-transformed cells, but this had no effect on cell-growth or stress-induced phospholipid signalling responses. Upon treatment with wortmannin, PtdIns3P levels were reduced by approximately 40% within 15 min in both cell lines. Stable expression of YFP-2xFYVE in Arabidopsis plants labelled different subcellular structures in root compared with shoot tissues. In addition labelling the motile structures common to all cells, YFP-2xFYVE strongly labelled the vacuolar membrane in leaf epidermal and guard cells, suggesting that cell differentiation alters the distribution of PtdIns3P. In dividing BY-2 cells, YFP-2xFYVE-labelled vesicles surrounded the newly formed cell plate, suggesting a role for PtdIns3P in cytokinesis. Together, these data show that YFP-2xFYVE may be used as a biosensor to specifically visualize PtdIns3P in living plant cells. © 2006 The Authors.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Wiley Blackwell Publishing, Inc
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
CONFOCAL LASER SCANNING MICROSCOPY
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FYVE DOMAIN
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LIPID SIGNALLING
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VESICLE TRAFFICKING
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YELLOW FLUORESCENT PROTEIN
dc.subject.classification
Biología Celular, Microbiología
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Visualization of PtdIns3P dynamics in living plant cells
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2021-12-03T19:59:56Z
dc.journal.volume
47
dc.journal.number
5
dc.journal.pagination
687-700
dc.journal.pais
Reino Unido
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dc.journal.ciudad
Londres
dc.description.fil
Fil: Vermeer, Joop E.M.. University of Amsterdam; Países Bajos
dc.description.fil
Fil: Van Leeuwen, Wessel. University of Amsterdam; Países Bajos
dc.description.fil
Fil: Tobeña Santamaria, Rafa. University of Amsterdam; Países Bajos
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Fil: Laxalt, Ana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
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Fil: Jones, David R.. The Netherlands Cancer Institute; Países Bajos
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Fil: Divecha, Nullin. The Netherlands Cancer Institute; Países Bajos
dc.description.fil
Fil: Gadella Jr., Theodorus W.J.. University of Amsterdam; Países Bajos
dc.description.fil
Fil: Munnik, Teun. University of Amsterdam; Países Bajos
dc.journal.title
Plant Journal
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1111/j.1365-313X.2006.02830.x
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1111/j.1365-313X.2006.02830.x
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