title: Mechanisms underlying the protein-kinase mediated regulation of the HERG potassium channel synthesis.
abstract: The HERG (human ether-a-go-go related gene) potassium channel aids in the repolarization of the cardiomyocyte membrane at the end of each action potential. We have previously shown that sustained protein kinase A or C (PKA and PKC) activity specifically enhances channel synthesis over the course of hours to days in heterologous expression and cardiac myocytes. The kinase-mediated augmentation of the channel is post-transcriptional and occurs near or at the endoplasmic reticulum. Here we report our further investigations into the mechanisms of kinase-mediated augmentation of HERG channel protein. We show that HERG channel phosphorylation alone is not sufficient for the PKA-dependent increase to occur. In vitro translation studies indicate that an additional factor is required for the process. Pharmacologic inhibitors suggest that the channel augmentation is not due to kinase-mediated alteration in proteasome or lysosome activity. PKA activation had no effect on stability of HERG mRNA and polyribosomal profiling showed that kinase activity did not elevate translation from low to high rates. Transcriptional inhibition results suggest that the additional cellular factor is a PKA-regulated protein. Together, these findings suggest that PKA-mediated augmentation of HERG abundance is more complex than previously appreciated involving enhancement of already active translation rates, phosphorylation of the channel protein and at least one other cyclic-AMP/PKA-responsive protein. Further exploration of molecular components of this regulatory pathway will be necessary to determine exact mechanism and the biomedical impact of this process in vivo.
authors.affiliation: Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY, USA.
authors.forename: Yamini
authors.initials: Y
authors.lastname: Krishnan
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Renjian
authors.initials: R
authors.lastname: Zheng
authors.forename: Vikram
authors.initials: V
authors.lastname: Kanda
authors.forename: Thomas V
authors.initials: TV
authors.lastname: McDonald
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism; D000242:Cyclic AMP / Q000502:physiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism* / Q000502:physiology; D003609:Dactinomycin / Q000494:pharmacology; D000072237:ERG1 Potassium Channel; D004789:Enzyme Activation; D051638:Ether-A-Go-Go Potassium Channels / Q000096:biosynthesis / Q000235:genetics*; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D010766:Phosphorylation; D061988:Proteasome Inhibitors / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D048948:14-3-3 Proteins; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; C000606913:KCNH2 protein, human; D007976:Leupeptins; D061988:Proteasome Inhibitors; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D003609:Dactinomycin; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde