title: Upregulation of proteasome activity rescues cardiomyocytes following pulse treatment with a proteasome inhibitor.
abstract: This study examined the hypothesis that cardiomyocyte metabolism is inherently linked to the Ubiquitin Proteasome System. Rat neonatal ventricular cardiomyocytes were pulse-treated with 5 αM lactacystin for 30 min, resulting in 95% loss of proteasome activity, and then maintained in culture for up to 24 h. Pulse-treatment resulted in 36% decrease in cardiomyocyte mitochondrial reductase activity by 8 h which improved to 15% by 24 h. Bax proteins were increased 2.5-fold by 8 h but declined by 16 h. Similar effects were observed for ubiquitinated proteins suggesting recovery of proteasome function. Proteasome activity started to increase by 4 h and was back to baseline by 16 h. Multiple proteasome subunits, including α1, were upregulated with peak 2 to 2.5-fold increased protein levels at 8-16 h post-lactacystin which then declined. Incubating cardiomyocytes with 4 αM morpholino-antisense oligonucleotides to the α1-subunit for up to 24 h post-lactacystin diminished recovery of proteasome activity (45% at 24 h) and prevented the increase in α1 protein levels. Ubiquitinated proteins remained elevated and cardiomyocyte mitochondrial reductase activity was decreased 35% by 16 h. These results show that diminished function of the ubiquitin proteasome system decreases cardiomyocyte metabolism. If proteasome activity recovers, function improves, but preventing recovery diminishes metabolic function supporting the hypothesis that cardiomyocyte metabolism is inherently linked to the ubiquitin proteasome system.
authors.affiliation: The Center for Heart and Lung Research, The Feinstein Institute for Medical Research Manhasset, NY, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.affiliation: The Center for Heart and Lung Research, The Feinstein Institute for Medical Research Manhasset, NY, USA ; Department of Biology, LIU-POST Brookville, NY, USA.
authors.forename: Justine
authors.initials: J
authors.lastname: Calise
authors.affiliation: The Center for Heart and Lung Research, The Feinstein Institute for Medical Research Manhasset, NY, USA.
authors.forename: Keren
authors.initials: K
authors.lastname: Powell
authors.affiliation: The Center for Heart and Lung Research, The Feinstein Institute for Medical Research Manhasset, NY, USA.
authors.forename: Andras
authors.initials: A
authors.lastname: Divald
authors.affiliation: The Center for Heart and Lung Research, The Feinstein Institute for Medical Research Manhasset, NY, USA ; Departments of Science Education and Molecular Medicine, Hofstra-NSLIJ School of Medicine Hempstead, NY.
authors.forename: Saul R
authors.initials: SR
authors.lastname: Powell
keywords: Ubiquitin proteasome system; apoptosis; cardiomyocyte; metabolic function