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
Main Authors: Lacalle, Rosa Ana, Gómez-Moutón, Concepción, Barber, Domingo F, Jiménez-Baranda, Sonia, Mira, Emilia, Martínez-A, Carlos, Carrera, Ana C, Mañes, Santos
Format: Journal Article
Language:English
Published: England 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.
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
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  givenname: Sonia
  surname: Jiménez-Baranda
  fullname: Jiménez-Baranda, Sonia
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  givenname: Emilia
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  fullname: Mira, Emilia
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  fullname: Martínez-A, Carlos
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  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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StartPage 6207
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
URI https://www.ncbi.nlm.nih.gov/pubmed/15564381
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