%0 Journal Article
%T Direct and heterologous approaches to identify the LET-756/FGF interactome
%A Cornel Popovici
%A Yael Berda
%A Fabien Conchonaud
%A Aur¨¦lie Harbis
%A Daniel Birnbaum
%A R¨¦gine Roubin
%J BMC Genomics
%D 2006
%I BioMed Central
%R 10.1186/1471-2164-7-105
%X To identify nuclear factors associated with LET-756, we used three approaches. First, we screened a two-hybrid cDNA library derived from mixed stages worms and from a normalized library, using LET-756 as bait. This direct approach allowed the identification of several binding partners that play various roles in the nucleus/nucleolus, such as PAL-1, a transcription regulator, or RPS-16, a component of the small ribosomal subunit. The interactions were validated by co-immunoprecipitation and determination of their site of occurrence in mammalian cells. Second, because patterns of protein interactions may be conserved throughout species, we searched for orthologs of known mammalian interactors and measured binary interaction with these predicted candidates. We found KIN-3 and KIN-10, the orthologs of CK2¦Á and CK2¦Â, as new partners of LET-756. Third, following the assumption that recognition motifs mediating protein interaction may be conserved between species, we screened a two-hybrid cDNA human library using LET-756 as bait. Among the few FGF partners detected was 14-3-3¦Â. In support of this interaction we showed that the two 14-3-3¦Â orthologous proteins, FTT-1 and FTT-2/PAR-5, interacted with LET-756.We have conducted the first extensive search for LET-756 interactors using a multi-directional approach and established the first interaction map of LET-756/FGF with other FGF binding proteins from other species. The interactors identified play various roles in developmental process or basic biochemical events such as ribosome biogenesis.FGFs constitute a superfamily of pleiotropic growth factors involved in multiple cellular processes such as mitogenesis, angiogenesis and mesoderm induction [1]. There are 22 FGFs in humans. Except FGF11-14, they exert their biological activities by acting as extracellular growth factors binding to receptors (FGFR1-4) of the tyrosine kinase receptor superfamily [2]. In addition, FGF1-3 and FGF11-14 are localized in the nucleus and functi
%U http://www.biomedcentral.com/1471-2164/7/105