title: Gelsolin interacts with LamR, hnRNP U, nestin, Arp3 and β-tubulin in human melanoma cells as revealed by immunoprecipitation and mass spectrometry.
abstract: Gelsolin, a multifunctional actin binding protein, plays a not yet fully understood role in tumorigenesis. Therefore the goal of this study was to identify additional molecular partners of gelsolin in human melanoma cells, separately in the cytoplasmic compartment and cell nuclei. For this purpose we performed immunoprecipitation experiments based on a modified protocol followed by mass spectrometry. The obtained results were confirmed by Western blot analysis, proximity ligation assays and confocal microscopy. As expected gelsolin interacted with actin, in particular we demonstrate its interaction with cytoplasmic β and γ actins, and a newly discovered actin isoform, actbl2. As new gelsolin-interacting partners we identified the ribosomal protein Rpsa, also known as a non-integrin laminin receptor (LamR), and the heterogeneous nuclear ribonucleoprotein hnRNP U. Our data furthermore indicate that gelsolin interacts with particular components of the three cytoskeleton systems: nestin (intermediate filaments), Arp3 (actin cytoskeleton) and β-tubulin (microtubules). We also report for the first time that gelsolin is a constituent of midbodies, a tubulin containing structure formed at the end of cytokinesis.
authors.affiliation: Department of Cell Pathology, Faculty of Biotechnology, University of Wroclaw, ul. Joliot-Curie 14a, 50-383 Wrocław, Poland. Electronic address: antonina.mazur@uwr.edu.pl.
authors.forename: Antonina Joanna
authors.initials: AJ
authors.lastname: Mazur
authors.affiliation: Department of Cell Pathology, Faculty of Biotechnology, University of Wroclaw, ul. Joliot-Curie 14a, 50-383 Wrocław, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Radaszkiewicz
authors.affiliation: Department of Cell Pathology, Faculty of Biotechnology, University of Wroclaw, ul. Joliot-Curie 14a, 50-383 Wrocław, Poland.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Makowiecka
authors.affiliation: Department of Cell Pathology, Faculty of Biotechnology, University of Wroclaw, ul. Joliot-Curie 14a, 50-383 Wrocław, Poland.
authors.forename: Maria
authors.initials: M
authors.lastname: Malicka-Błaszkiewicz
authors.affiliation: Department of Anatomy and Molecular Embryology, Faculty of Medicine, Ruhr-University, Universitätsstr. 150, 44780 Bochum, Germany.
authors.forename: Hans Georg
authors.initials: HG
authors.lastname: Mannherz
authors.affiliation: Department of Cell Pathology, Faculty of Biotechnology, University of Wroclaw, ul. Joliot-Curie 14a, 50-383 Wrocław, Poland.
authors.forename: Dorota
authors.initials: D
authors.lastname: Nowak
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D051378:Actin-Related Protein 3 / Q000378:metabolism; D015153:Blotting, Western; D045744:Cell Line, Tumor; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D018260:Gelsolin / Q000378:metabolism*; D034722:Heterogeneous-Nuclear Ribonucleoprotein U / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D008545:Melanoma / Q000378:metabolism*; D018613:Microscopy, Confocal; D064231:Nestin / Q000378:metabolism; D017474:Receptors, Laminin / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012269:Ribosomal Proteins; D014162:Transfection; D014404:Tubulin / Q000378:metabolism
chemical_list: C495928:ACTR3 protein, human; D051378:Actin-Related Protein 3; D003598:Cytoskeletal Proteins; D018260:Gelsolin; D034722:Heterogeneous-Nuclear Ribonucleoprotein U; C577484:NES protein, human; D064231:Nestin; C510624:RPSA protein, human; D017474:Receptors, Laminin; D012269:Ribosomal Proteins; D014404:Tubulin
keywords: Gelsolin; LamR; Nestin; actbl2; hnRNP U; β Tubulin,