title: CEACAM1 functionally interacts with filamin A and exerts a dual role in the regulation of cell migration.
abstract: The carcinoembryonic antigen-related cell adhesion molecule CEACAM1 (CD66a) and the scaffolding protein filamin A have both been implicated in tumor cell migration. In the present study we identified filamin A as a novel binding partner for the CEACAM1-L cytoplasmic domain in a yeast two-hybrid screen. Direct binding was shown by surface plasmon resonance analysis and by affinity precipitation assays. The association was shown for human and rodent CEACAM1-L in endogenous CEACAM1-L expressing cells. To address functional aspects of the interaction, we used a well-established melanoma cell system. We found in different migration studies that the interaction of CEACAM1-L and filamin A drastically reduced migration and cell scattering, whereas each of these proteins when expressed alone, acted promigratory. CEACAM1-L binding to filamin A reduced the interaction of the latter with RalA, a member of the Ras-family of GTPases. Furthermore, co-expression of CEACAM1-L and filamin A led to a reduced focal adhesion turnover. Independent of the presence of filamin A, the expression of CEACAM1-L led to an increased phosphorylation of focal adhesions and to altered cytoskeletal rearrangements during monolayer wound healing assays. Together, our data demonstrate a novel mechanism for how CEACAM1-L regulates cell migration via its interaction with filamin A.
authors.affiliation: Institut für Biochemie und Molekularbiologie, Charité, Universitätsmedizin Berlin, Campus Benjamin Franklin, 14195 Berlin, Germany.
authors.forename: Esther
authors.initials: E
authors.lastname: Klaile
authors.forename: Mario M
authors.initials: MM
authors.lastname: Müller
authors.forename: Christoph
authors.initials: C
authors.lastname: Kannicht
authors.forename: Bernhard B
authors.initials: BB
authors.lastname: Singer
authors.forename: Lothar
authors.initials: L
authors.lastname: Lucka
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology*; D003285:Contractile Proteins / Q000235:genetics / Q000502:physiology*; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D064448:Filamins; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000502:physiology*; D051419:Paxillin / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D051381:Rats; D020798:Two-Hybrid System Techniques; D014443:Tyrosine / Q000187:drug effects / Q000378:metabolism
chemical_list: D015703:Antigens, CD; C069847:CD66 antigens; D015815:Cell Adhesion Molecules; D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins; D051419:Paxillin; D014443:Tyrosine; D051416:Focal Adhesion Protein-Tyrosine Kinases