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J Biol Chem. 2016 Oct 7;291(41):21510-21518. Epub 2016 Aug 16.

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

Author information

1
From the Departments of Chemistry and.
2
From the Departments of Chemistry and vegli001@umn.edu.
3
Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455.
4
From the Departments of Chemistry and bowser@umn.edu.

Abstract

The sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) and phospholamban (PLN) complex regulates heart relaxation through its removal of cytosolic Ca2+ 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 (Kd <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.

KEYWORDS:

Phospholamban (PLN); RNA-protein interaction; calcium ATPase (SERCA); cardiomyopathy; fluorescence anisotropy; microRNA (miRNA); oligonucleotide analogs; protein complex; protein-nucleic acid interaction; structure-function

PMID:
27531746
PMCID:
PMC5076822
DOI:
10.1074/jbc.M116.738807
[Indexed for MEDLINE]
Free PMC Article

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