Recovery of mitogenic activity of a growth factor mutant with a nuclear translocation sequence

Science. 1990 Sep 28;249(4976):1567-70. doi: 10.1126/science.1699274.

Abstract

Heparin-binding growth factor-1 (HBGF-1) is an angiogenic polypeptide mitogen for mesoderm- and neuroectoderm-derived cells in vitro and remains biologically active after truncation of the amino-terminal domain (HBGF-1 alpha) of the HBGF-1 beta precursor. Polymerase chain reaction mutagenesis and prokaryotic expression systems were used to prepare a mutant of HBGF-1 alpha lacking a putative nuclear translocation sequence (amino acid residues 21 to 27; HBGF-1U). Although HBGF-1U retains its ability to bind to heparin, HBGF-1U fails to induce DNA synthesis and cell proliferation at concentrations sufficient to induce intracellular receptor-mediated tyrosine phosphorylation and c-fos expression. Attachment of the nuclear translocation sequence from yeast histone 2B at the amino terminus of HBGF-1U yields a chimeric polypeptide (HBGF-1U2) with mitogenic activity in vitro and indicates that nuclear translocation is important for this biological response.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Binding, Competitive
  • Cattle
  • Cell Division / drug effects
  • Cell Line
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • DNA Replication / drug effects
  • Endothelium, Vascular / drug effects
  • Endothelium, Vascular / metabolism
  • Fibroblast Growth Factor 1 / genetics*
  • Fibroblast Growth Factor 1 / metabolism
  • Fibroblast Growth Factor 1 / pharmacology
  • Kinetics
  • Mice
  • Mitogens / pharmacology
  • Molecular Sequence Data
  • Mutation*
  • Oligonucleotide Probes
  • Receptors, Mitogen / metabolism
  • Receptors, Vascular Endothelial Growth Factor
  • Recombinant Proteins / metabolism
  • Recombinant Proteins / pharmacology
  • Transcription, Genetic / drug effects

Substances

  • Mitogens
  • Oligonucleotide Probes
  • Receptors, Mitogen
  • Recombinant Proteins
  • Fibroblast Growth Factor 1
  • Receptors, Vascular Endothelial Growth Factor