Isolation of proteins that interact with the signal transduction molecule Dof and identification of a functional domain conserved between Dof and vertebrate BCAP.
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.
Battersby
Alysia
A
Institut für Genetik, Universität zu Köln, Weyertal 121, D-50931 Köln, Germany.
Csiszár
Agnes
A
Leptin
Maria
M
Wilson
Robert
R
D048868:Adaptor Proteins, Signal Transducing; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002478:Cells, Cultured; D017124:Conserved Sequence; D019281:Dimerization; D004330:Drosophila / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029721:Drosophila Proteins; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
D048868:Adaptor Proteins, Signal Transducing; C447641:BANK1 protein, human; D002352:Carrier Proteins; D029721:Drosophila Proteins; D019476:Insect Proteins; D008565:Membrane Proteins; C487977:PIK3AP1 protein, human; D012333:RNA, Messenger; D017468:Receptors, Fibroblast Growth Factor; C115576:stumps protein, Drosophila