Live-cell imaging of phosphatidic acid dynamics in pollen tubes visualized by Spo20p-derived biosensor

© 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

Bibliographische Detailangaben
Veröffentlicht in:The New phytologist. - 1990. - 203(2014), 2 vom: 09. Juli, Seite 483-494
1. Verfasser: Potocký, Martin (VerfasserIn)
Weitere Verfasser: Pleskot, Roman, Pejchar, Přemysl, Vitale, Nicolas, Kost, Benedikt, Žárský, Viktor
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2014
Zugriff auf das übergeordnete Werk:The New phytologist
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Nicotiana tabacum (tobacco) Spo20p live-cell microscopy phosphatidic acid (PA) phospholipase D pollen tube Diglycerides Phosphatidic Acids mehr... Phosphatidylinositol 4,5-Diphosphate Qb-SNARE Proteins Qc-SNARE Proteins Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Spo20 protein, S cerevisiae Phospholipase D EC 3.1.4.4
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100 1 |a Potocký, Martin  |e verfasserin  |4 aut 
245 1 0 |a Live-cell imaging of phosphatidic acid dynamics in pollen tubes visualized by Spo20p-derived biosensor 
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520 |a © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust. 
520 |a Although phosphatidic acid (PA) is structurally the simplest membrane phospholipid, it has been implicated in the regulation of many cellular events, including cytoskeletal dynamics, membrane trafficking and stress responses. Plant PA shows rapid turnover but the information about its spatio-temporal distribution in plant cells is missing. Here we demonstrate the use of a lipid biosensor that enables us to monitor PA dynamics in plant cells. The biosensor consists of a PA-binding domain of yeast SNARE Spo20p fused to fluorescent proteins. Live-cell imaging of PA dynamics in transiently transformed tobacco (Nicotiana tabacum) pollen tubes was performed using confocal laser scanning microscopy. In growing pollen tubes, PA shows distinct annulus-like fluorescence pattern in the plasma membrane behind the extreme tip. Coexpression studies with markers for other plasmalemma signaling lipids phosphatidylinositol 4,5-bisphosphate and diacylglycerol revealed limited colocalization at the shoulders of the apex. PA distribution and concentrations show distinct responses to various lipid signaling inhibitors. Fluorescence recovery after photobleaching (FRAP) analysis suggests high PA turnover in the plasma membrane. Our data show that a biosensor based on the Spo20p-PA binding domain is suitable for live-cell imaging of PA also in plant cells. In tobacco pollen tubes, distinct subapical PA maximum corroborates its involvement in the regulation of endocytosis and actin dynamics 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a Nicotiana tabacum (tobacco) 
650 4 |a Spo20p 
650 4 |a live-cell microscopy 
650 4 |a phosphatidic acid (PA) 
650 4 |a phospholipase D 
650 4 |a pollen tube 
650 7 |a Diglycerides  |2 NLM 
650 7 |a Phosphatidic Acids  |2 NLM 
650 7 |a Phosphatidylinositol 4,5-Diphosphate  |2 NLM 
650 7 |a Qb-SNARE Proteins  |2 NLM 
650 7 |a Qc-SNARE Proteins  |2 NLM 
650 7 |a Recombinant Fusion Proteins  |2 NLM 
650 7 |a Saccharomyces cerevisiae Proteins  |2 NLM 
650 7 |a Spo20 protein, S cerevisiae  |2 NLM 
650 7 |a Phospholipase D  |2 NLM 
650 7 |a EC 3.1.4.4  |2 NLM 
700 1 |a Pleskot, Roman  |e verfasserin  |4 aut 
700 1 |a Pejchar, Přemysl  |e verfasserin  |4 aut 
700 1 |a Vitale, Nicolas  |e verfasserin  |4 aut 
700 1 |a Kost, Benedikt  |e verfasserin  |4 aut 
700 1 |a Žárský, Viktor  |e verfasserin  |4 aut 
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773 1 8 |g volume:203  |g year:2014  |g number:2  |g day:09  |g month:07  |g pages:483-494 
856 4 0 |u http://dx.doi.org/10.1111/nph.12814  |3 Volltext 
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