Analysis of endogenous and exogenous nuclear translocation of fibroblast growth factor-1 in NIH 3T3 cells

Biochem Biophys Res Commun. 1992 Nov 16;188(3):982-91. doi: 10.1016/0006-291x(92)91328-n.

Abstract

Nuclear localization of fibroblast growth factors (FGF) have been reported by many laboratories. We demonstrate here that FGF-1, the precursor for acidic FGF contains a putative nuclear translocation sequence (NTS) NYKKPKL, which is able to direct the expression of the bacterial beta galactosidase (beta gal) gene to the nucleus of transfected NIH 3T3 cells. However, this NTS is unable to target either FGF-1 itself or a FGF-1-beta gal fusion protein into the nucleus, suggesting that FGF-1 may contain an additional sequence which prevents endogenously expressed FGF-1 from being translocated into the nucleus. Indeed, when FGF-1 was fused to the NTS derived from the yeast histone 2B gene, the chimeric construct also failed to be transported into the nucleus either by itself or as a beta gal fusion protein. Interestingly, when 125I-FGF-1 was used to stimulate quiescent NIH 3T3 cells, a significant amount of internalized 125I-FGF-1 (approximately 10%) was found within the nucleus and the nuclear localization of FGF-1 through the exogenous pathway could be significantly reduced by suramin, an inhibitor of the interaction of FGF-1 with its receptor. These data suggest that while FGF-1 contains a NTS, nuclear translocation requires an exogenous and not an endogenous pathway.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Biological Transport
  • Cell Nucleus / metabolism*
  • Cloning, Molecular
  • Cytosol / metabolism
  • Fibroblast Growth Factor 1 / isolation & purification
  • Fibroblast Growth Factor 1 / metabolism*
  • Immunohistochemistry
  • Mice
  • Molecular Sequence Data
  • Protein Precursors / metabolism*
  • Protein Sorting Signals
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • beta-Galactosidase / isolation & purification
  • beta-Galactosidase / metabolism

Substances

  • Protein Precursors
  • Protein Sorting Signals
  • Recombinant Fusion Proteins
  • Fibroblast Growth Factor 1
  • beta-Galactosidase