Coordinated Localization and Antagonistic Function of NtPLC3 and PI4P 5-Kinases in the Subapical Plasma Membrane of Tobacco Pollen Tubes
Polar tip growth of pollen tubes is regulated by the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P<sub>2</sub>), which localizes in a well-defined region of the subapical plasma membrane. How the PtdIns(4,5)P<sub>2</sub> region is maintained is cur...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-04-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/9/4/452 |
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| author | Irene Stenzel Till Ischebeck Linh Hai Vu-Becker Mara Riechmann Praveen Krishnamoorthy Marta Fratini Ingo Heilmann |
| author_facet | Irene Stenzel Till Ischebeck Linh Hai Vu-Becker Mara Riechmann Praveen Krishnamoorthy Marta Fratini Ingo Heilmann |
| author_sort | Irene Stenzel |
| collection | DOAJ |
| description | Polar tip growth of pollen tubes is regulated by the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P<sub>2</sub>), which localizes in a well-defined region of the subapical plasma membrane. How the PtdIns(4,5)P<sub>2</sub> region is maintained is currently unclear. In principle, the formation of PtdIns(4,5)P<sub>2</sub> by PI4P 5-kinases can be counteracted by phospholipase C (PLC), which hydrolyzes PtdIns(4,5)P<sub>2</sub>. Here, we show that fluorescence-tagged tobacco NtPLC3 displays a subapical plasma membrane distribution which frames that of fluorescence-tagged PI4P 5-kinases, suggesting that NtPLC3 may modulate PtdIns(4,5)P<sub>2</sub>-mediated processes in pollen tubes. The expression of a dominant negative NtPLC3 variant resulted in pollen tube tip swelling, consistent with a delimiting effect on PtdIns(4,5)P<sub>2</sub> production. When pollen tube morphologies were assessed as a quantitative read-out for PtdIns(4,5)P<sub>2</sub> function, NtPLC3 reverted the effects of a coexpressed PI4P 5-kinase, demonstrating that NtPLC3-mediated breakdown of PtdIns(4,5)P<sub>2</sub> antagonizes the effects of PtdIns(4,5)P<sub>2</sub> overproduction in vivo. When analyzed by spinning disc microscopy, fluorescence-tagged NtPLC3 displayed discontinuous membrane distribution omitting punctate areas of the membrane, suggesting that NtPLC3 is involved in the spatial restriction of plasma membrane domains also at the nanodomain scale. Together, the data indicate that NtPLC3 may contribute to the spatial restriction of PtdIns(4,5)P<sub>2</sub> in the subapical plasma membrane of pollen tubes. |
| first_indexed | 2024-03-10T20:41:34Z |
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| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T20:41:34Z |
| publishDate | 2020-04-01 |
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| series | Plants |
| spelling | doaj.art-fc0e91bfcc994d15b0f02bdffe0352042023-11-19T20:39:29ZengMDPI AGPlants2223-77472020-04-019445210.3390/plants9040452Coordinated Localization and Antagonistic Function of NtPLC3 and PI4P 5-Kinases in the Subapical Plasma Membrane of Tobacco Pollen TubesIrene Stenzel0Till Ischebeck1Linh Hai Vu-Becker2Mara Riechmann3Praveen Krishnamoorthy4Marta Fratini5Ingo Heilmann6Department of Cellular Biochemistry, Institute for Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyDepartment of Plant Biochemistry, Albrecht-von-Haller-Institute of Plant Sciences and Göttingen Center for Molecular Biosciences (GZMB), Georg August University Göttingen, 37077 Göttingen, GermanyDepartment of Cellular Biochemistry, Institute for Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyDepartment of Cellular Biochemistry, Institute for Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyDepartment of Cellular Biochemistry, Institute for Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyDepartment of Cellular Biochemistry, Institute for Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyDepartment of Cellular Biochemistry, Institute for Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyPolar tip growth of pollen tubes is regulated by the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P<sub>2</sub>), which localizes in a well-defined region of the subapical plasma membrane. How the PtdIns(4,5)P<sub>2</sub> region is maintained is currently unclear. In principle, the formation of PtdIns(4,5)P<sub>2</sub> by PI4P 5-kinases can be counteracted by phospholipase C (PLC), which hydrolyzes PtdIns(4,5)P<sub>2</sub>. Here, we show that fluorescence-tagged tobacco NtPLC3 displays a subapical plasma membrane distribution which frames that of fluorescence-tagged PI4P 5-kinases, suggesting that NtPLC3 may modulate PtdIns(4,5)P<sub>2</sub>-mediated processes in pollen tubes. The expression of a dominant negative NtPLC3 variant resulted in pollen tube tip swelling, consistent with a delimiting effect on PtdIns(4,5)P<sub>2</sub> production. When pollen tube morphologies were assessed as a quantitative read-out for PtdIns(4,5)P<sub>2</sub> function, NtPLC3 reverted the effects of a coexpressed PI4P 5-kinase, demonstrating that NtPLC3-mediated breakdown of PtdIns(4,5)P<sub>2</sub> antagonizes the effects of PtdIns(4,5)P<sub>2</sub> overproduction in vivo. When analyzed by spinning disc microscopy, fluorescence-tagged NtPLC3 displayed discontinuous membrane distribution omitting punctate areas of the membrane, suggesting that NtPLC3 is involved in the spatial restriction of plasma membrane domains also at the nanodomain scale. Together, the data indicate that NtPLC3 may contribute to the spatial restriction of PtdIns(4,5)P<sub>2</sub> in the subapical plasma membrane of pollen tubes.https://www.mdpi.com/2223-7747/9/4/452Phosphatidylinositol 4,5-bisphosphatePtdIns(4,5)P<sub>2</sub>phospholipase CPI4P 5-kinasepollen tube tip swelling |
| spellingShingle | Irene Stenzel Till Ischebeck Linh Hai Vu-Becker Mara Riechmann Praveen Krishnamoorthy Marta Fratini Ingo Heilmann Coordinated Localization and Antagonistic Function of NtPLC3 and PI4P 5-Kinases in the Subapical Plasma Membrane of Tobacco Pollen Tubes Plants Phosphatidylinositol 4,5-bisphosphate PtdIns(4,5)P<sub>2</sub> phospholipase C PI4P 5-kinase pollen tube tip swelling |
| title | Coordinated Localization and Antagonistic Function of NtPLC3 and PI4P 5-Kinases in the Subapical Plasma Membrane of Tobacco Pollen Tubes |
| title_full | Coordinated Localization and Antagonistic Function of NtPLC3 and PI4P 5-Kinases in the Subapical Plasma Membrane of Tobacco Pollen Tubes |
| title_fullStr | Coordinated Localization and Antagonistic Function of NtPLC3 and PI4P 5-Kinases in the Subapical Plasma Membrane of Tobacco Pollen Tubes |
| title_full_unstemmed | Coordinated Localization and Antagonistic Function of NtPLC3 and PI4P 5-Kinases in the Subapical Plasma Membrane of Tobacco Pollen Tubes |
| title_short | Coordinated Localization and Antagonistic Function of NtPLC3 and PI4P 5-Kinases in the Subapical Plasma Membrane of Tobacco Pollen Tubes |
| title_sort | coordinated localization and antagonistic function of ntplc3 and pi4p 5 kinases in the subapical plasma membrane of tobacco pollen tubes |
| topic | Phosphatidylinositol 4,5-bisphosphate PtdIns(4,5)P<sub>2</sub> phospholipase C PI4P 5-kinase pollen tube tip swelling |
| url | https://www.mdpi.com/2223-7747/9/4/452 |
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