PTEN regulates motility but not directionality during leukocyte chemotaxis
The localization at opposite cell poles of phosphatidylinositol-3 kinases and PTEN (phosphatase and tensin homolog on chromosome 10) governs Dictyostelium chemotaxis. To study this model in mammalian cells, we analyzed the dynamic redistribution of green fluorescent protein (GFP)-tagged PTEN chimera...
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Published in: | Journal of cell science Vol. 117; no. Pt 25; pp. 6207 - 6215 |
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01-12-2004
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Abstract | The localization at opposite cell poles of phosphatidylinositol-3 kinases and PTEN (phosphatase and tensin homolog on chromosome 10) governs Dictyostelium chemotaxis. To study this model in mammalian cells, we analyzed the dynamic redistribution of green fluorescent protein (GFP)-tagged PTEN chimeras during chemotaxis. N- or C-terminus GFP-tagged PTEN was distributed homogeneously in the cytoplasm of chemotaxing PTEN-negative Jurkat cells and PTEN-positive HL60 cells. Moreover, we did not detect uropod accumulation of endogenous PTEN in chemoattractant-stimulated HL60 cells. Cell fractionation indicated that both endogenous and ectopically expressed PTEN were confined largely to the cytosol, and that chemoattractant stimulation did not alter this location. PTEN re-expression in Jurkat cells or PTEN depletion by specific siRNA in HL60 cells did not affect cell gradient sensing; PTEN nonetheless modulated chemoattractant-induced actin polymerization and the speed of cell movement. The results suggest a role for PTEN in regulating actin polymerization, but not directionality during mammalian cell chemotaxis. |
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AbstractList | The localization at opposite cell poles of phosphatidylinositol-3 kinases and PTEN (phosphatase and tensin homolog on chromosome 10) governs Dictyostelium chemotaxis. To study this model in mammalian cells, we analyzed the dynamic redistribution of green fluorescent protein (GFP)-tagged PTEN chimeras during chemotaxis. N- or C-terminus GFP-tagged PTEN was distributed homogeneously in the cytoplasm of chemotaxing PTEN-negative Jurkat cells and PTEN-positive HL60 cells. Moreover, we did not detect uropod accumulation of endogenous PTEN in chemoattractant-stimulated HL60 cells. Cell fractionation indicated that both endogenous and ectopically expressed PTEN were confined largely to the cytosol, and that chemoattractant stimulation did not alter this location. PTEN re-expression in Jurkat cells or PTEN depletion by specific siRNA in HL60 cells did not affect cell gradient sensing; PTEN nonetheless modulated chemoattractant-induced actin polymerization and the speed of cell movement. The results suggest a role for PTEN in regulating actin polymerization, but not directionality during mammalian cell chemotaxis. The localization at opposite cell poles of phosphatidylinositol-3 kinases and PTEN (phosphatase and tensin homolog on chromosome 10) governs Dictyostelium chemotaxis. To study this model in mammalian cells, we analyzed the dynamic redistribution of green fluorescent protein (GFP)-tagged PTEN chimeras during chemotaxis. N- or C-terminus GFP-tagged PTEN was distributed homogenously in the cytoplasm of chemotaxing PTEN-negative Jurkat cells and PTEN-positive HL60 cells. Moreover, we did not detect uropod accumulation of endogenous PTEN in chemoattractant-stimulated HL60 cells. Cell fractionation indicated that both endogenous and ectopically expressed PTEN were confined largely to the cytosol, and that chemoattractant stimulation did not alter this location. PTEN re-expression in Jurkat cells or PTEN depletion by specific siRNA in HL60 cells did not affect cell gradient sensing; PTEN nonetheless modulated chemoattractant-induced actin polymerization and the speed of cell movement. The results suggest a role for PTEN in regulating actin polymerization, but not directionality during mammalian cell chemotaxis. |
Author | Jiménez-Baranda, Sonia Carrera, Ana C Mañes, Santos Gómez-Moutón, Concepción Martínez-A, Carlos Lacalle, Rosa Ana Mira, Emilia Barber, Domingo F |
Author_xml | – sequence: 1 givenname: Rosa Ana surname: Lacalle fullname: Lacalle, Rosa Ana organization: Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, UAM Campus de Cantoblanco, 28049 Madrid, Spain – sequence: 2 givenname: Concepción surname: Gómez-Moutón fullname: Gómez-Moutón, Concepción – sequence: 3 givenname: Domingo F surname: Barber fullname: Barber, Domingo F – sequence: 4 givenname: Sonia surname: Jiménez-Baranda fullname: Jiménez-Baranda, Sonia – sequence: 5 givenname: Emilia surname: Mira fullname: Mira, Emilia – sequence: 6 givenname: Carlos surname: Martínez-A fullname: Martínez-A, Carlos – sequence: 7 givenname: Ana C surname: Carrera fullname: Carrera, Ana C – sequence: 8 givenname: Santos surname: Mañes fullname: Mañes, Santos |
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Snippet | The localization at opposite cell poles of phosphatidylinositol-3 kinases and PTEN (phosphatase and tensin homolog on chromosome 10) governs Dictyostelium... |
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SubjectTerms | Actins - metabolism Blotting, Western Catalysis Cell Movement Chemotactic Factors - pharmacology Chemotaxis Cloning, Molecular Cytoplasm - metabolism Cytosol - metabolism Dictyostelium DNA, Complementary - metabolism Dose-Response Relationship, Drug Green Fluorescent Proteins - metabolism HL-60 Cells Humans Jurkat Cells Leukocytes - cytology Membrane Microdomains - metabolism Microscopy, Fluorescence Microscopy, Video Phosphatidylinositol 3-Kinases - metabolism Phosphoric Monoester Hydrolases - metabolism Phosphoric Monoester Hydrolases - physiology Protein Structure, Tertiary PTEN Phosphohydrolase RNA, Small Interfering - metabolism Subcellular Fractions - metabolism Time Factors Tumor Suppressor Proteins - metabolism Tumor Suppressor Proteins - physiology |
Title | PTEN regulates motility but not directionality during leukocyte chemotaxis |
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