title: Phosphorylation of radixin regulates cell polarity and Mrp-2 distribution in hepatocytes.
abstract: Radixin, the dominant ezrin-radixin-moesin (ERM) protein in hepatocytes, has two important binding domains: an NH(2)-terminal region that binds to plasma membrane and a COOH-terminal region that binds to F-actin after a conformational activation by phosphorylation at Thr564. The present studies were undertaken to investigate the cellular changes in expression of radixin in WIF-B cells and to assess radixin distribution and its influence on cell polarity. We used a recombinant adenoviral expression system encoding radixin wild-type and Thr564 mutants fused to cyan fluorescent protein (CFP), as well as conventional immunostaining procedures. Functional analyses were characterized quantitatively. Similar to endogenous radixin, adenovirus-infected radixin-CFP-wild type and nonphosphorylatable radixin-CFP-T564A were found to be expressed heavily in the compartment of canalicular membrane vacuoles, typically colocalizing with multidrug resistance-associated protein 2 (Mrp-2). Expression of radixin-CFP-T564D, which mimics constant phosphorylation, was quite different, being rarely associated with canalicular membranes. The WIF-B cells were devoid of a secretory response, T567D radixin became predominantly redistributed to the basolateral membrane, usually in the form of dense, long spikes and fingerlike projections, and the altered cell polarity involved changes in apical membrane markers. Differences in polar distribution of radixin suggest a role for the linker protein in promoting formation and plasticity of membrane surface projections and also suggest that radixin might be an organizer and regulator of Mrp-2 and cell polarity in hepatocytes.
authors.affiliation: Keck School of Medicine, Division of Gastrointestinal & Liver Diseases, Univ. of Southern California, Zonal Ave., HMR 101, Los Angeles, CA 90033, USA.
authors.forename: Jo
authors.initials: J
authors.lastname: Suda
authors.forename: Lixin
authors.initials: L
authors.lastname: Zhu
authors.forename: Serhan
authors.initials: S
authors.lastname: Karvar
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D016764:Cell Polarity / Q000502:physiology*; D022081:Cell Surface Extensions / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D006825:Hybridomas; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000378:metabolism*; D050505:Mutant Proteins / Q000235:genetics; D010766:Phosphorylation / Q000235:genetics; D051381:Rats; D014162:Transfection; D014617:Vacuoles / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D050505:Mutant Proteins; C000715087:ABCC2 protein, human; C071776:radixin