Activation of Wiskott-Aldrich syndrome protein and its association with other proteins by stromal cell-derived factor-1alpha is associated with cell migration in a T-lymphocyte line.
OBJECTIVE
Chemokines play a central role in lymphocyte trafficking and homing. The actin cytoskeleton is involved in cell morphological changes and motility. Wiskott-Aldrich syndrome (WAS) protein (WASP) has been implicated in regulation of cytoskeleton rearrangement. To evaluate mechanisms that might be involved in migration of T cells, we examined effects of stromal cell-derived factor (SDF)-1alpha on WASP and associated proteins.


METHODS
Jurkat T cells were stimulated by SDF-1alpha and analyzed for chemotaxis and also by Western blot analysis for signal transduction.


RESULTS
Jurkat T cells displayed chemotaxis to SDF-1alpha, which was inhibited by pretreatment of cells with either pertussis toxin, a Galphai protein inhibitor, wortmannin or Ly294002, phophatidylinositol 3-kinase inhibitors, or herbimycin, a protein tyrosine kinase inhibitor. WASP was tyrosine phosphorylated in response to SDF-1alpha stimulation in Jurkat T cells. Crk associated substrate (Cas), Nck, and focal adhesion kinase (FAK) were also phosphorylated after SDF-1alpha stimulation. Moreover, activated Nck interacted with Cas and WASP as determined by co-immunoprecipitation, and FAK also bound to Cas.


CONCLUSIONS
These data suggest that WASP, Cas, Nck, and FAK may play a role in SDF-1alpha-induced migration of the T-cell line, Jurkat.
Department of Microbiology/Immunology and the Walther Oncology Center, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Seiichi
S
Okabe
Seiji
S
Fukuda
Hal E
HE
Broxmeyer
D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing; D001704:Biopolymers; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000494:pharmacology / Q000502:physiology*; D002633:Chemotaxis / Q000187:drug effects / Q000502:physiology*; D050719:Crk-Associated Substrate Protein; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure; D004791:Enzyme Inhibitors / Q000494:pharmacology; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006801:Humans; D019169:Jurkat Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D046911:Macromolecular Substances; D009363:Neoplasm Proteins / Q000378:metabolism; D015513:Oncogene Proteins / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D019718:Receptors, CXCR4 / Q000187:drug effects / Q000502:physiology; D011994:Recombinant Proteins / Q000494:pharmacology; D050717:Retinoblastoma-Like Protein p130; D015398:Signal Transduction / Q000187:drug effects; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism; D051316:Wiskott-Aldrich Syndrome Protein
D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D001704:Biopolymers; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D050719:Crk-Associated Substrate Protein; D004791:Enzyme Inhibitors; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D046911:Macromolecular Substances; D009363:Neoplasm Proteins; D015513:Oncogene Proteins; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010750:Phosphoproteins; D011505:Protein-Tyrosine Kinases; D011506:Proteins; D019718:Receptors, CXCR4; D011994:Recombinant Proteins; D050717:Retinoblastoma-Like Protein p130; D051316:Wiskott-Aldrich Syndrome Protein; C493841:BCAR1 protein, human; C515255:CXCL12 protein, human; C078332:Nck protein; C495767:WAS protein, human; C496050:PTK2 protein, human