Rheostatic Regulation of the SERCA/Phospholamban Membrane Protein Complex Using Non-Coding RNA and Single-Stranded DNA oligonucleotides

Soller, Kailey J.; Verardi, Raffaello; Meng, Jing; Abrol, Neha; Yang, Jing; Walsh, Naomi; Vostrikov, Vitaly V.; Robia, Seth L.; Bowser, Michael T.; Veglia, Gianluigi

doi:10.1038/srep13000

Cited by 9 publications

(14 citation statements)

References 51 publications

(73 reference statements)

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“…As RNA length increases, inhibition is gradually relieved, with the 50-mer re-establishing SERCA basal activity. This is similar to the incremental relief we have previously observed for ssDNA (12). The random sequence library that was used in these experiments confirms that the RNA-PLN interaction is sequence-independent.…”

Section: Rna Sequences Bind Phospholamban With Low Nanomolar K Dsupporting

confidence: 72%

“…In the current study, we report that many XNAs bind PLN with similar strong affinity to what was found previously for SPIDRs (12). These molecules, which are tunable by length, would allow clinicians to match the reversal of SERCA inhibition to the severity of the disease.…”

mentioning

confidence: 57%

“…Reversing SERCA Inhibition-Previously, we found that ssDNA of varying lengths could relieve the inhibition of SERCA by PLN to different extents (12). We also determined that a 50-mer of RNA bound PLN with similar affinity and relieved inhibition of SERCA to a similar extent as ssDNA (12).…”

Section: Rna Sequences Bind Phospholamban With Low Nanomolar K Dmentioning

confidence: 99%

“…We also determined that a 50-mer of RNA bound PLN with similar affinity and relieved inhibition of SERCA to a similar extent as ssDNA (12). In the current study, we determined the full extent of different lengths of RNA binding to PLN and its functional effects on the SERCA-PLN complex.…”

Section: Rna Sequences Bind Phospholamban With Low Nanomolar K Dmentioning

confidence: 99%

“…We previously reported that random sequence, singlestranded DNA and RNA bind to PLN with high affinity (K d Ͻ 10 nM), and more importantly, relieve its inhibitory effects on SERCA, restoring the enzyme's basal activity. We found that PLN remained bound to SERCA upon the addition of ssDNA, and that the effects of ssDNA on the SERCA-PLN complex are length-dependent and tunable (12). We propose to refer to these unique sequences as short, protein-interacting DNAs and RNAs (i.e.…”

mentioning

confidence: 99%

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Reversal of Phospholamban Inhibition of the Sarco(endo)plasmic Reticulum Ca2+-ATPase (SERCA) Using Short, Protein-interacting RNAs and Oligonucleotide Analogs

Soller¹,

Yang²,

Veglia

et al. 2016

Journal of Biological Chemistry

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The sarco(endo)plasmic reticulum Ca 2؉ -ATPase (SERCA) and phospholamban (PLN) complex regulates heart relaxation through its removal of cytosolic Ca 2؉ during diastole. Dysfunction of this complex has been related to many heart disorders and is therefore a key pharmacological target. There are currently no therapeutics that directly target either SERCA or PLN. It has been previously reported that single-stranded DNA binds PLN with strong affinity and relieves inhibition of SERCA in a length-dependent manner. In the current article, we demonstrate that RNAs and single-stranded oligonucleotide analogs, or xeno nucleic acids (XNAs), also bind PLN strongly (K d <10 nM) and relieve inhibition of SERCA. Affinity for PLN is sequence-independent. Relief of PLN inhibition is length-dependent, allowing SERCA activity to be restored incrementally. The improved in vivo stability of XNAs offers more realistic pharmacological potential than DNA or RNA. We also found that microRNAs (miRNAs) 1 and 21 bind PLN strongly and relieve PLN inhibition of SERCA to a greater extent than a similar length random sequence RNA mixture. This may suggest that miR-1 and miR-21 have evolved to contain distinct sequence elements that are more effective at relieving PLN inhibition than random sequences.Calcium cycling in cardiomyocytes is a tightly regulated process that ensures proper muscle contractility (1, 2). Many of the proteins involved in this cycle have been implicated in cardiac failure, the leading cause of death world-wide (3, 4). Given the complexity of heart failure phenotypes, strategies for reversing declining cardiac performance are diverse and necessary. Gene therapy efforts for inherited forms of heart disease have been growing, but with the recent failure of the Calcium Upregulation by Percutaneous Administration of Gene Therapy in Cardiac Disease (CUPID) trials, drug development targeting specific proteins remains an essential, complimentary effort (5-7). A therapeutic that is tunable to specific phenotypes would be ideal for reversing aberrant calcium cycling.The sarco(endo)plasmic reticulum Ca 2ϩ -ATPase (SERCA) 3 is an essential Ca 2ϩ -handling protein that removes Ca 2ϩ ions from the cytosol, causing the heart muscle to relax. SERCA is a P-type ATPase, which in human cardiomyocytes is responsible for the removal of ϳ70% of Ca 2ϩ from the cytosol (8). This process, coupled with other Ca 2ϩ transport mechanisms, lowers the cytosolic Ca 2ϩ concentration enough to allow for muscle relaxation (diastole) (1). Phospholamban (PLN) is a 52-amino acid, single-pass transmembrane protein that inhibits SERCA when not phosphorylated by lowering its apparent calcium affinity and hampering Ca 2ϩ transport into the sarcoplasmic reticulum (9, 10). Upon ␤-adrenergic stimulation, protein kinase A (PKA) will phosphorylate PLN at Ser-16, restoring the apparent calcium affinity and Ca 2ϩ transport of SERCA (11). We previously reported that random sequence, singlestranded DNA and RNA bind to PLN with high affinity (K d Ͻ 10 nM), and more import...

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Section: Rna Sequences Bind Phospholamban With Low Nanomolar K Dsupporting

confidence: 72%

mentioning

confidence: 57%