Expression of autocrine motility factor/phosphohexose isomerase in Cos7 cells.
Autocrine motility factor (AMF) is identical to the glycolytic enzyme phosphohexose isomerase (PHI) and overexpression of AMF/PHI is associated with tumor malignancy. In order to study the overexpression of AMF/PHI, an HA-tagged AMF construct was transiently transfected into Cos7 cells. Expression of a tagged AMF-HA allowed us to determine that over a period of 16 hours only a small amount (0.1-1%) of total cellular AMF-HA was secreted into the cell medium. Cell-associated AMF-HA was exclusively cytosolic as it could be completely extracted with Triton X-100 and concentrated within actin rich pseudopodial domains. Treatment of the cells with the glycolysis inhibitor oxamate disrupted the association of AMF-HA with actin concentrations demonstrating that glycolysis regulates the formation of these AMF/PHI-associated actin-rich protrusions. AMF/PHI is a well-characterized tumor cell secreted cytokine and we identify here an alternate intracellular function for this glycolytic enzyme/cytokine in cell motility.
Lagana
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Department of Pathology and Cell Biology, Université de Montréal, Montreal, Quebec, H3C 3J7, Canada.
Duchaine
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Raz
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DesGroseillers
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Nabi
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IR
D000199:Actins / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D002465:Cell Movement; D017077:Culture Media, Conditioned / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D019281:Dimerization; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects; D005956:Glucose-6-Phosphate Isomerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006019:Glycolysis / Q000187:drug effects; D007770:L-Lactate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism; D008970:Molecular Weight; D017830:Octoxynol / Q000378:metabolism; D010072:Oxamic Acid / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D011554:Pseudopodia / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D019289:Pyruvic Acid / Q000378:metabolism / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
D000199:Actins; D017077:Culture Media, Conditioned; D011993:Recombinant Fusion Proteins; D019289:Pyruvic Acid; D017830:Octoxynol; D007770:L-Lactate Dehydrogenase; D005956:Glucose-6-Phosphate Isomerase; D010072:Oxamic Acid