title: Rheostatic Regulation of the SERCA/Phospholamban Membrane Protein Complex Using Non-Coding RNA and Single-Stranded DNA oligonucleotides.
abstract: The membrane protein complex between sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) and phospholamban (PLN) is a prime therapeutic target for reversing cardiac contractile dysfunctions caused by calcium mishandling. So far, however, efforts to develop drugs specific for this protein complex have failed. Here, we show that non-coding RNAs and single-stranded DNAs (ssDNAs) interact with and regulate the function of the SERCA/PLN complex in a tunable manner. Both in HEK cells expressing the SERCA/PLN complex, as well as in cardiac sarcoplasmic reticulum preparations, these short oligonucleotides bind and reverse PLN's inhibitory effects on SERCA, increasing the ATPase's apparent Ca(2+) affinity. Solid-state NMR experiments revealed that ssDNA interacts with PLN specifically, shifting the conformational equilibrium of the SERCA/PLN complex from an inhibitory to a non-inhibitory state. Importantly, we achieved rheostatic control of SERCA function by modulating the length of ssDNAs. Since restoration of Ca(2+) flux to physiological levels represents a viable therapeutic avenue for cardiomyopathies, our results suggest that oligonucleotide-based drugs could be used to fine-tune SERCA function to counterbalance the extent of the pathological insults.
authors: Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Kailey J
authors: KJ
authors: Soller
authors: Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Raffaello
authors: R
authors: Verardi
authors: Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Meng
authors: M
authors: Jing
authors: Department of Cell and Molecular Physiology, Stritch School of Medicine, Loyola University Chicago, 60153.
authors: Neha
authors: N
authors: Abrol
authors: Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Jing
authors: J
authors: Yang
authors: Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Naomi
authors: N
authors: Walsh
authors: Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Vitaly V
authors: VV
authors: Vostrikov
authors: Department of Cell and Molecular Physiology, Stritch School of Medicine, Loyola University Chicago, 60153.
authors: Seth L
authors: SL
authors: Robia
authors: Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Michael T
authors: MT
authors: Bowser
authors: Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455.|Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Gianluigi
authors: G
authors: Veglia
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002470:Cell Survival; D004277:DNA, Single-Stranded / Q000378:metabolism*; D000939:Epitopes / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000378:metabolism*; D009841:Oligonucleotides / Q000378:metabolism*; D011485:Protein Binding; D022661:RNA, Untranslated / Q000378:metabolism*; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism*; D034421:Sus scrofa; D000099211:Phospholamban
chemical_list: D000596:Amino Acids; D002135:Calcium-Binding Proteins; D004277:DNA, Single-Stranded; D000939:Epitopes; D008565:Membrane Proteins; D009841:Oligonucleotides; D022661:RNA, Untranslated; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000099211:Phospholamban