Endogenous basic fibroblast growth factor isoforms involved in different intracellular protein complexes.
Four forms of basic fibroblast growth factor (bFGF or FGF-2) result from an alternative initiation of translation involving one AUG (155-amino acid form) and three CUGs (210-, 201- and 196-amino acid forms). These different forms of bFGF show different intracellular biological activities. To identify their intracellular targets, the 210- and 155-amino acid forms of bFGF were independently transfected into CHO cells and their correct subcellular localizations were verified, the 155-amino acid bFGF form being essentially cytoplasmic whereas the 210-amino acid protein was nuclear. The radiation fragmentation method was used to determine the target size of the different bFGF isoforms in the transfected CHO cells and to show that the 210- and 155-amino acids bFGF isoforms were included in protein complexes of 320 and 130 kDa respectively. Similar results were obtained using the SK-Hep1 cell line, which naturally expressed all forms of bFGF. Co-immunoprecipitation assays using different chimaeric bFGF-chloramphenicol acetyltransferase proteins showed that different cellular proteins are associated with different parts of the bFGF molecule. We conclude that bFGF isoforms are involved in different molecular complexes in the cytosol and nucleus, which would reflect different functions for these proteins.
Patry
V
V
INSERM U397, Institut Louis Bugnard, CHU Rangueil, Toulouse, France.
Bugler
B
B
Maret
A
A
Potier
M
M
Prats
H
H
D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D006528:Carcinoma, Hepatocellular; D006224:Cricetinae; D004307:Dose-Response Relationship, Radiation; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D005720:Gamma Rays; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D007424:Intracellular Fluid / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D007536:Isomerism; D046911:Macromolecular Substances; D011233:Precipitin Tests; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects; D014162:Transfection; D014407:Tumor Cells, Cultured
D046911:Macromolecular Substances; D016222:Fibroblast Growth Factor 2