Efficient ADAM22 surface expression is mediated by phosphorylation-dependent interaction with 14-3-3 protein family members.
ADAM22 is one of three catalytically inactive ADAM family members highly expressed in the brain. ADAM22 has numerous splice variants, all with considerable cytoplasmic tails of up to 148 amino acids. ADAM22 can act to inhibit cell proliferation, however, it has been suggested that it also acts as an adhesion protein. We identified three 14-3-3 protein members by a yeast two-hybrid screen and show by co-immunoprecipitation that the cytoplasmic domain of ADAM22 can interact with all six 14-3-3 proteins expressed in the brain. In addition, we show that 14-3-3 proteins interact preferentially with the serine phosphorylated precursor form of ADAM22. ADAM22 has two 14-3-3 protein binding consensus motifs; the first binding site, spanning residues 831-834, was shown to be the most crucial for 14-3-3 binding to occur. The interaction between ADAM22 and 14-3-3 proteins is dependent on phosphorylation of ADAM22, but not of 14-3-3 proteins. ADAM22 point mutants lacking functional 14-3-3 protein binding motifs could no longer accumulate efficiently at the cell surface. Deletion of both 14-3-3 binding sites and newly identified ER retention motifs restored localization of ADAM22 at the cell surface. These results reveal a role for 14-3-3 proteins in targeting ADAM22 to the membrane by masking ER retention signals.
Gödde
Nathan J
NJ
Department of Surgery, University of Melbourne, Parkville 3050, Australia.
D'Abaco
Giovanna M
GM
Paradiso
Lucy
L
Novak
Ulrike
U
D048948:14-3-3 Proteins / Q000235:genetics / Q000378:metabolism*; D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005910:Glioma / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D007668:Kidney / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D017354:Point Mutation; D011485:Protein Binding; D012441:Saccharomyces cerevisiae; D020798:Two-Hybrid System Techniques
D048948:14-3-3 Proteins; D009419:Nerve Tissue Proteins; D051722:ADAM Proteins; C498474:ADAM22 protein, human