Focal adhesion kinase signaling is decreased in polyamine-depleted IEC-6 cells.
Polyamines are essential to the migration of epithelial cells in the intestinal mucosa. Cells depleted of polyamines do not attach as rapidly to the extracellular matrix and do not form the actin stress fibers essential for migration. Because both attachment and stress fiber formation depend on integrin signaling and the formation of focal adhesions, we examined these and related processes in polyamine-depleted IEC-6 cells. There was general decreased tyrosine phosphorylation of focal adhesion kinase (FAK), and, specifically, decreased phosphorylation of Tyr-925, the paxillin binding site. In control cells, FAK phosphorylation was rapid after attachment to the extracellular matrix, while attached cells depleted of polyamines had significantly delayed phosphorylation. FAK activity was also significantly inhibited in polyamine-depleted cells as was the phosphorylation of paxillin. Polyamine-depleted cells failed to spread normally after attachment, and immunocytochemistry showed little colocalization of FAK and actin compared with controls. Focal adhesion complex formation was greatly reduced in the absence of polyamines. These data suggest that defective integrin signaling may, at least in part, account for the decreased rates of attachment, actin stress fiber formation, spreading, and migration observed in polyamine-depleted cells.
Ray
R M
RM
Department of Physiology, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.
Viar
M J
MJ
McCormack
S A
SA
Johnson
L R
LR
D000199:Actins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D003598:Cytoskeletal Proteins / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases; D007150:Immunohistochemistry; D016023:Integrins / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D051419:Paxillin; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011073:Polyamines / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*; D014018:Tissue Distribution
D000199:Actins; D003598:Cytoskeletal Proteins; D016023:Integrins; D051419:Paxillin; D010750:Phosphoproteins; D011073:Polyamines; D011505:Protein-Tyrosine Kinases; D051416:Focal Adhesion Protein-Tyrosine Kinases