Isolation of proteins that interact with the signal transduction molecule Dof and identification of a functional domain conserved between Dof and vertebrate BCAP

J Mol Biol. 2003 Jun 6;329(3):479-93. doi: 10.1016/s0022-2836(03)00489-3.

Abstract

Dof is a large molecule essential for signal transduction by the two FGF receptors in Drosophila. It contains two ankyrin repeats and a coiled-coil region, but has no other recognisable structural motif. Dof shares these features with its closest vertebrate relatives, the B-cell signalling molecules BCAP and BANK. In addition, this family of proteins shares a region of homology upstream of the ankyrin repeats, which we call the Dof/BCAP/BANK (DBB) motif. We have identified 44 proteins that interact with Dof in a yeast two-hybrid screen. These include the Drosophila FGF-receptor Heartless and Dof itself. We show that the integrity of the DBB motif is required both for Dof and for BCAP to form dimers. Analysis of the interactions between a set of deletion constructs of Dof and the panel of interactors suggests that Dof may adopt different conformations, with a folded conformation stabilized by interactions between the DBB motif and the C-terminal part of the protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Carrier Proteins / chemistry*
  • Carrier Proteins / metabolism
  • Cells, Cultured
  • Conserved Sequence*
  • Dimerization
  • Drosophila / chemistry
  • Drosophila / genetics
  • Drosophila / metabolism
  • Drosophila Proteins*
  • Gene Expression Regulation, Developmental
  • In Situ Hybridization
  • Insect Proteins / chemistry*
  • Insect Proteins / genetics
  • Insect Proteins / metabolism*
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism
  • Molecular Sequence Data
  • Protein Binding
  • Protein Structure, Tertiary
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Receptors, Fibroblast Growth Factor / metabolism
  • Signal Transduction*
  • Two-Hybrid System Techniques

Substances

  • Adaptor Proteins, Signal Transducing
  • BANK1 protein, human
  • Carrier Proteins
  • Drosophila Proteins
  • Insect Proteins
  • Membrane Proteins
  • PIK3AP1 protein, human
  • RNA, Messenger
  • Receptors, Fibroblast Growth Factor
  • stumps protein, Drosophila