Phosphorylation-regulated nucleocytoplasmic trafficking of internalized fibroblast growth factor-1.
Fibroblast growth factor-1 (FGF-1), which stimulates cell growth, differentiation, and migration, is capable of crossing cellular membranes to reach the cytosol and the nucleus in cells containing specific FGF receptors. The cell entry process can be monitored by phosphorylation of the translocated FGF-1. We present evidence that phosphorylation of FGF-1 occurs in the nucleus by protein kinase C (PKC)delta. The phosphorylated FGF-1 is subsequently exported to the cytosol. A mutant growth factor where serine at the phosphorylation site is exchanged with glutamic acid, to mimic phosphorylated FGF-1, is constitutively transported to the cytosol, whereas a mutant containing alanine at this site remains in the nucleus. The export can be blocked by leptomycin B, indicating active and receptor-mediated nuclear export of FGF-1. Thapsigargin, but not leptomycin B, prevents the appearance of active PKCdelta in the nucleus, and FGF-1 is in this case phosphorylated in the cytosol. Leptomycin B increases the amount of phosphorylated FGF-1 in the cells by preventing dephosphorylation of the growth factor, which seems to occur more rapidly in the cytoplasm than in the nucleus. The nucleocytoplasmic trafficking of the phosphorylated growth factor is likely to play a role in the activity of internalized FGF-1.
Wiedłocha
Antoni
A
Institute for Cancer Research, The Norwegian Radium Hospital, 0310 Oslo, Norway. antoni.wiedlocha@labmed.uio.no
Nilsen
Trine
T
Wesche
Jørgen
J
Sørensen
Vigdis
V
Małecki
Jedrzej
J
Marcinkowska
Ewa
E
Olsnes
Sjur
S
D019943:Amino Acid Substitution; D000818:Animals; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000378:metabolism*; D016895:Culture Media, Serum-Free; D003593:Cytoplasm / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis; D004072:Digitonin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D016220:Fibroblast Growth Factor 1 / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism; D006367:HeLa Cells; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D007202:Indicators and Reagents / Q000494:pharmacology; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D041681:NIH 3T3 Cells; D010766:Phosphorylation; D014176:Protein Biosynthesis; D011493:Protein Kinase C / Q000032:analysis / Q000378:metabolism; D051745:Protein Kinase C-delta; D013347:Subcellular Fractions; D013462:Sulfur Radioisotopes / Q000378:metabolism; D019284:Thapsigargin / Q000494:pharmacology; D014158:Transcription, Genetic; D014357:Trypsin / Q000494:pharmacology
D016895:Culture Media, Serum-Free; D007202:Indicators and Reagents; D013462:Sulfur Radioisotopes; D016220:Fibroblast Growth Factor 1; D018698:Glutamic Acid; D019284:Thapsigargin; D006493:Heparin; D004247:DNA; D008715:Methionine; C497259:Prkcd protein, mouse; C497258:PRKCD protein, human; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D014357:Trypsin; D004072:Digitonin