title: miR-200c dampens cancer cell migration via regulation of protein kinase A subunits.
abstract: Expression of miR-200c is a molecular switch to determine cellular fate towards a mesenchymal or epithelial phenotype. miR-200c suppresses the early steps of tumor progression by preventing epithelial-mesenchymal transition (EMT) and intravasation of tumor cells. Unraveling the underlying molecular mechanisms might pinpoint to novel therapeutic options. To better understand these mechanisms it is crucial to identify targets of miR-200c. Here, we employ a combination of quantitative proteomic and bioinformatic strategies to identify novel miR-200c targets. We identify and confirm two subunits of the central cellular kinase protein kinase A (PKA), namely PRKAR1A and PRKACB, to be directly regulated by miR-200c. Notably, siRNA-mediated downregulation of both proteins phenocopies the migratory behavior of breast cancer cells after miR-200c overexpression. Patient data from publicly accessible databases supports a miR-200c-PKA axis. Thus, our study identifies the PKA heteroprotein as an important mediator of miR-200c induced repression of migration in breast cancer cells. By bioinformatics, we define a miRNA target cluster consisting of PRKAR1A, PRKAR2B, PRKACB, and COF2, which is targeted by a group of 14 miRNAs.
authors.affiliation: Institute of Molecular Medicine and Cell Research, Albert-Ludwigs-University Freiburg, Freiburg, Germany.|Faculty of Biology, Albert-Ludwigs-University Freiburg, Freiburg, Germany.
authors.forename: Florian Christoph
authors.initials: FC
authors.lastname: Sigloch
authors.affiliation: Institute of Molecular Medicine and Cell Research, Albert-Ludwigs-University Freiburg, Freiburg, Germany.|BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany.
authors.forename: Ulrike Christina
authors.initials: UC
authors.lastname: Burk
authors.affiliation: Institute of Molecular Medicine and Cell Research, Albert-Ludwigs-University Freiburg, Freiburg, Germany.
authors.forename: Martin Lothar
authors.initials: ML
authors.lastname: Biniossek
authors.affiliation: Experimental Medicine I, Nikolaus-Fiebiger-Center for Molecular Medicine, University Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Brabletz
authors.affiliation: Institute of Molecular Medicine and Cell Research, Albert-Ludwigs-University Freiburg, Freiburg, Germany.|BIOSS Centre for Biological Signaling Studies, University of Freiburg, Freiburg, Germany.|German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Schilling
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D002853:Chromatography, Liquid; D054751:Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / Q000096:biosynthesis / Q000235:genetics*; D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit / Q000096:biosynthesis / Q000235:genetics*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D054461:Lim Kinases / Q000235:genetics; D035683:MicroRNAs / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D053719:Tandem Mass Spectrometry
chemical_list: D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; C517834:PRKAR1A protein, human; D034741:RNA, Small Interfering; C515892:LIMK1 protein, human; D054461:Lim Kinases; D054751:Cyclic AMP-Dependent Protein Kinase Catalytic Subunits; C517760:PRKACB protein, human
keywords: PKA; cofilin; miR-200c; miRNA target cluster; proteomics