caa a22 4500 475744837 CHVBK 20180406123510.0 cr unu---uuuuu 170329e20000801xx s 000 0 eng 10.1023/A:1026507727700 doi (NATIONALLICENCE)springer-10.1023/A:1026507727700 Translation is regulated via the 3′ untranslated region of α-myosin heavy chain mRNA by calcium but not by its localization [Elektronische Daten] [Gordana Nikcevic, Merja Perhonen, Samuel Boateng, Brenda Russell] Posttranscriptional regulation plays an important role in α-myosin heavy chain (α-MyHC) protein synthesis in cardiac muscle cells. In the present study, we test the effects of calcium and mRNA mislocalization on α-MyHC translation in order to determine the mechanism(s) contributing to translational block via the 3′ untranslated region (3′UTR). Neonatal rat cardiac myocytes were treated for 6 h with L-isoproterenol (10 μM) to enhance beating, with 10 μM verapamil to block beating and mislocalize mRNA, or with 3 μM colchicine to enhance beating but mislocalize mRNA by depolymerization of the microtubules. In order to determine whether translation is regulated by the 3′UTR, either a control (SV40 3′UTR) or the experimental (α-MyHC 3′UTR) was placed after a luciferase reporter gene and transfected into the myocytes. The amount of luciferase protein only decreased significantly in verapamil arrested cells transfected with the α-MyHC 3′UTR construct (P < 0.01). To control for the possibility that pharmacological treatments might affect transcription or message stability, we analyzed neomycin and luciferase mRNA levels transcribed from the same transfected plasmid. No significant changes were found with an RNase protection assay. These results suggest that calcium but not mRNA localization regulates protein synthesis and further, this is mediated by the 3′ untranslated region of α-MyHC. Kluwer Academic Publishers, 2000 Nikcevic Gordana Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, 60612-7342, Chicago, IL, USA aut Perhonen Merja Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, 60612-7342, Chicago, IL, USA aut Boateng Samuel Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, 60612-7342, Chicago, IL, USA aut Russell Brenda Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, 60612-7342, Chicago, IL, USA aut Journal of Muscle Research & Cell Motility Kluwer Academic Publishers 21/6(2000-08-01), 599-607 0142-4319 21:6<599 2000 21 10974 https://doi.org/10.1023/A:1026507727700 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1026507727700 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nikcevic Gordana Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, 60612-7342, Chicago, IL, USA aut NATIONALLICENCE 700 1- Perhonen Merja Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, 60612-7342, Chicago, IL, USA aut NATIONALLICENCE 700 1- Boateng Samuel Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, 60612-7342, Chicago, IL, USA aut NATIONALLICENCE 700 1- Russell Brenda Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, 60612-7342, Chicago, IL, USA aut NATIONALLICENCE 773 0- Journal of Muscle Research & Cell Motility Kluwer Academic Publishers 21/6(2000-08-01), 599-607 0142-4319 21:6<599 2000 21 10974 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer