PI3-kinase signaling contributes to orientation in shallow gradients and enhances speed in steep chemoattractant gradients.
Dictyostelium cells that chemotax towards cAMP produce phosphatidylinositol (3,4,5)-trisphosphate [PtdIns(3,4,5)P(3)] at the leading edge, which has been implicated in actin reorganization and pseudopod extension. However, in the absence of PtdIns(3,4,5)P(3) signaling, cells will chemotax via alternative pathways. Here we examined the potential contribution of PtdIns(3,4,5)P(3) to chemotaxis of wild-type cells. The results show that steep cAMP gradients (larger than 10% concentration difference across the cell) induce strong PtdIns(3,4,5)P(3) patches at the leading edge, which has little effect on the orientation but strongly enhances the speed of the cell. Using a new sensitive method for PtdIns(3,4,5)P(3) detection that corrects for the volume of cytosol in pixels at the boundary of the cell, we show that, in shallow cAMP gradient (less than 5% concentration difference across the cell), PtdIns(3,4,5)P(3) is still somewhat enriched at the leading edge. Cells lacking PI3-kinase (PI3K) activity exhibit poor chemotaxis in these shallow gradients. Owing to the reduced speed and diminished orientation of the cells in steep and shallow gradients, respectively, cells lacking PtdIns(3,4,5)P(3) signaling require two- to six-fold longer times to reach a point source of chemoattractant compared with wild-type cells. These results show that, although PI3K signaling is dispensable for chemotaxis, it gives the wild type an advantage over mutant cells.
Department of Molecular Cell Biology, University of Groningen, Kerklaan 30, 9751NN Haren, The Netherlands.
Leonard
L
Bosgraaf
Ineke
I
Keizer-Gunnink
Peter J M
PJ
Van Haastert
D000818:Animals; D002465:Cell Movement; D002630:Chemotactic Factors / Q000378:metabolism*; D002633:Chemotaxis; D000242:Cyclic AMP / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004023:Dictyostelium / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018715:Microscopy, Video / Q000379:methods; D008954:Models, Biological; D015233:Models, Statistical; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism* / Q000502:physiology; D021381:Protein Transport; D015398:Signal Transduction
D002630:Chemotactic Factors; D049452:Green Fluorescent Proteins; D000242:Cyclic AMP; D019869:Phosphatidylinositol 3-Kinases