| Autor: |
Hupfeld J; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany., Ernst M; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany., Knyrim M; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany., Binas S; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany., Kloeckner U; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany., Rabe S; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany., Quarch K; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany., Misiak D; Institute of Molecular Medicine, Medical Faculty of the Martin Luther University Halle-Wittenberg, Charles Tanford Protein Center, 06120 Halle (Saale), Germany., Fuszard M; Zentrum für Medizinische Grundlagenforschung, Core Facility-Proteomic Mass Spectrometry, Proteinzentrum Charles Tanford, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), Germany., Grossmann C; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany., Gekle M; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany., Schreier B; Julius-Bernstein Institute of Physiology, Medical Faculty of the Martin Luther University Halle-Wittenberg, 06112 Halle (Saale), Germany. |
| Abstrakt: |
MicroRNAs (miRs) contribute to different aspects of cardiovascular pathology, among them cardiac hypertrophy and atrial fibrillation. Cardiac miR expression was analyzed in a mouse model with structural and electrical remodeling. Next-generation sequencing revealed that miR-208b-3p was ~25-fold upregulated. Therefore, the aim of our study was to evaluate the impact of miR-208b on cardiac protein expression. First, an undirected approach comparing whole RNA sequencing data to miR-walk 2.0 miR-208b 3'-UTR targets revealed 58 potential targets of miR-208b being regulated. We were able to show that miR-208b mimics bind to the 3' untranslated region (UTR) of voltage-gated calcium channel subunit alpha1 C and Kcnj5, two predicted targets of miR-208b. Additionally, we demonstrated that miR-208b mimics reduce GIRK1/4 channel-dependent thallium ion flux in HL-1 cells. In a second undirected approach we performed mass spectrometry to identify the potential targets of miR-208b. We identified 40 potential targets by comparison to miR-walk 2.0 3'-UTR, 5'-UTR and CDS targets. Among those targets, Rock2 and Ran were upregulated in Western blots of HL-1 cells by miR-208b mimics. In summary, miR-208b targets the mRNAs of proteins involved in the generation of cardiac excitation and propagation, as well as of proteins involved in RNA translocation (Ran) and cardiac hypertrophic response (Rock2). |
