Phosphatidic acid regulation of PIPKI is critical for actin cytoskeletal reorganization[S]
Type I phosphatidylinositol-4-phosphate 5-kinase (PIPKI) is the main enzyme generating the lipid second messenger phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2], which has critical functions in many cellular processes, such as cytoskeletal reorganization, membrane trafficking, and signal transduc...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2012-12-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S002222752041795X |
| _version_ | 1828938090657349632 |
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| author | Akua N. Roach Ziqing Wang Ping Wu Feng Zhang Robin B. Chan Yoshiya Yonekubo Gilbert Di Paolo Alemayehu A. Gorfe Guangwei Du |
| author_facet | Akua N. Roach Ziqing Wang Ping Wu Feng Zhang Robin B. Chan Yoshiya Yonekubo Gilbert Di Paolo Alemayehu A. Gorfe Guangwei Du |
| author_sort | Akua N. Roach |
| collection | DOAJ |
| description | Type I phosphatidylinositol-4-phosphate 5-kinase (PIPKI) is the main enzyme generating the lipid second messenger phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2], which has critical functions in many cellular processes, such as cytoskeletal reorganization, membrane trafficking, and signal transduction. All three members of the PIPKI family are activated by phosphatidic acid (PA). However, how PA regulates the activity and functions of PIPKI have not been fully elucidated. In this study, we identify a PA-binding site on PIPKIγ. Mutation of this site inhibited the PA-stimulated activity and membrane localization of PIPKIγ as well as the formation of actin comets and foci induced by PIPKIγ. We also demonstrate that phospholipase D (PLD) generates a pool of PA involved in PIPKIγ regulation by showing that PLD inhibitors blocked the membrane localization of PIPKIγ and its ability to induce actin cytoskeletal reorganization. Targeting the PIPKIγ PA-binding-deficient mutant to membranes by a membrane localization sequence failed to restore the actin reorganization activity of PIPKIγ, suggesting that PA binding is not only involved in recruiting PIPKIγ to membranes but also may induce a conformational change. Taken together, these results reveal a new molecular mechanism through which PA regulates PIPKI and provides direct evidence that PA is important for the localization and functions of PIPKI in intact cells. |
| first_indexed | 2024-12-14T02:36:03Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-14T02:36:03Z |
| publishDate | 2012-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-2e61fb2afb794d0facbb86125f141cc72022-12-21T23:20:07ZengElsevierJournal of Lipid Research0022-22752012-12-01531225982609Phosphatidic acid regulation of PIPKI is critical for actin cytoskeletal reorganization[S]Akua N. Roach0Ziqing Wang1Ping Wu2Feng Zhang3Robin B. Chan4Yoshiya Yonekubo5Gilbert Di Paolo6Alemayehu A. Gorfe7Guangwei Du8Department of Pharmacology and the Center for Developmental Genetics, Stony Brook University, Stony Brook, NY 11794Department of Integrative Biology & Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030; andDepartment of Integrative Biology & Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030; andDepartment of Integrative Biology & Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030; andDepartment of Pathology and Cell Biology, Taub Institute for Research on Alzheimer's Disease and the Aging Brain, Columbia University Medical Center, New York, NY 10032Department of Integrative Biology & Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030; andDepartment of Pathology and Cell Biology, Taub Institute for Research on Alzheimer's Disease and the Aging Brain, Columbia University Medical Center, New York, NY 10032Department of Integrative Biology & Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030; andTo whom correspondence should be addressed. e-mail: guangwei.du@uth.tmc.edu.; Department of Integrative Biology & Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030; andType I phosphatidylinositol-4-phosphate 5-kinase (PIPKI) is the main enzyme generating the lipid second messenger phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2], which has critical functions in many cellular processes, such as cytoskeletal reorganization, membrane trafficking, and signal transduction. All three members of the PIPKI family are activated by phosphatidic acid (PA). However, how PA regulates the activity and functions of PIPKI have not been fully elucidated. In this study, we identify a PA-binding site on PIPKIγ. Mutation of this site inhibited the PA-stimulated activity and membrane localization of PIPKIγ as well as the formation of actin comets and foci induced by PIPKIγ. We also demonstrate that phospholipase D (PLD) generates a pool of PA involved in PIPKIγ regulation by showing that PLD inhibitors blocked the membrane localization of PIPKIγ and its ability to induce actin cytoskeletal reorganization. Targeting the PIPKIγ PA-binding-deficient mutant to membranes by a membrane localization sequence failed to restore the actin reorganization activity of PIPKIγ, suggesting that PA binding is not only involved in recruiting PIPKIγ to membranes but also may induce a conformational change. Taken together, these results reveal a new molecular mechanism through which PA regulates PIPKI and provides direct evidence that PA is important for the localization and functions of PIPKI in intact cells.http://www.sciencedirect.com/science/article/pii/S002222752041795Xtype I phosphatidylinositol-4-phosphate 5-kinasephosphatidylinositol-4,5-bisphosphatephospholipase Dactin cometsactin foci |
| spellingShingle | Akua N. Roach Ziqing Wang Ping Wu Feng Zhang Robin B. Chan Yoshiya Yonekubo Gilbert Di Paolo Alemayehu A. Gorfe Guangwei Du Phosphatidic acid regulation of PIPKI is critical for actin cytoskeletal reorganization[S] Journal of Lipid Research type I phosphatidylinositol-4-phosphate 5-kinase phosphatidylinositol-4,5-bisphosphate phospholipase D actin comets actin foci |
| title | Phosphatidic acid regulation of PIPKI is critical for actin cytoskeletal reorganization[S] |
| title_full | Phosphatidic acid regulation of PIPKI is critical for actin cytoskeletal reorganization[S] |
| title_fullStr | Phosphatidic acid regulation of PIPKI is critical for actin cytoskeletal reorganization[S] |
| title_full_unstemmed | Phosphatidic acid regulation of PIPKI is critical for actin cytoskeletal reorganization[S] |
| title_short | Phosphatidic acid regulation of PIPKI is critical for actin cytoskeletal reorganization[S] |
| title_sort | phosphatidic acid regulation of pipki is critical for actin cytoskeletal reorganization s |
| topic | type I phosphatidylinositol-4-phosphate 5-kinase phosphatidylinositol-4,5-bisphosphate phospholipase D actin comets actin foci |
| url | http://www.sciencedirect.com/science/article/pii/S002222752041795X |
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