Natriuretic peptide receptor-3 gene (NPR3): Nonsynonymous polymorphism results in significant reduction in protein expression because of accelerated degradation

Naveen Luke Pereira, Dong Lin, Linda Pelleymounter, Irene Moon, Gail Stilling, Bruce W. Eckloff, Eric D Wieben, Margaret May Redfield, John C Jr. Burnett, Vivien C. Yee, Richard M Weinshilboum

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9 Citations (Scopus)

Abstract

Background-The primary role of natriuretic peptide receptor-3 (NPR3) or NPR-C is in the clearance of natriuretic peptides that play an important role in modulating intravascular volume and vascular tone. Genetic variation in NPR3 has been associated with variation in blood pressure and obesity. Despite the importance of NPR3, sequence variation in the gene has not been addressed using DNA from different ethnic populations. We set out to identify and functionally characterize genetic variation in NPR3 in 3 ethnic groups. Methods and Results-DNA samples from 96 European American, 96 African American, and 96 Han Chinese American healthy subjects were used to resequence NPR3 exons, splice junctions, and flanking regions. We identified 105 polymorphisms, 50 of which were novel, including 8 nonsynonymous single-nucleotide polymorphisms, 7 were novel. Expression constructs were created for the nonsynonymous single-nucleotide polymorphisms. HEK293 cells were transfected with constructs for wild type and variant allozymes; and recombinant proteins were measured by quantitative Western blot analysis. The most significant change in NPR3 protein was observed for the Arg146 variant allozyme, with 20% of wild-type protein, primarily because of autophagy-dependent degradation. NPR3 structural modeling confirmed that the Arg146 variant protein was not compatible with wild-type conformation and could result in protein misfolding or instability. Conclusions-Multiple novel NPR3 genetic polymorphisms were identified in 3 ethnic groups. The Arg146 allozyme displayed a significant decrease in protein quantity because of degradation mediated predominantly by autophagy. This genetic variation could have a significant effect on the metabolism of natriuretic peptides with potential clinical implications.

Original languageEnglish (US)
Pages (from-to)201-210
Number of pages10
JournalCirculation: Cardiovascular Genetics
Volume6
Issue number2
DOIs
StatePublished - Apr 2013

Fingerprint

Natriuretic Peptides
Peptide Receptors
Genes
Proteins
Isoenzymes
Autophagy
Ethnic Groups
Single Nucleotide Polymorphism
Asian Americans
HEK293 Cells
DNA
Genetic Polymorphisms
Recombinant Proteins
African Americans
Blood Vessels
Exons
Healthy Volunteers
Obesity
Western Blotting
Blood Pressure

Keywords

  • Natriuretic peptide receptor-3
  • Natriuretic peptides
  • Pharmacogenetics
  • Polymorphism

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Genetics(clinical)
  • Genetics

Cite this

@article{439e1bf3aacb44178478c5e08d5ba15a,
title = "Natriuretic peptide receptor-3 gene (NPR3): Nonsynonymous polymorphism results in significant reduction in protein expression because of accelerated degradation",
abstract = "Background-The primary role of natriuretic peptide receptor-3 (NPR3) or NPR-C is in the clearance of natriuretic peptides that play an important role in modulating intravascular volume and vascular tone. Genetic variation in NPR3 has been associated with variation in blood pressure and obesity. Despite the importance of NPR3, sequence variation in the gene has not been addressed using DNA from different ethnic populations. We set out to identify and functionally characterize genetic variation in NPR3 in 3 ethnic groups. Methods and Results-DNA samples from 96 European American, 96 African American, and 96 Han Chinese American healthy subjects were used to resequence NPR3 exons, splice junctions, and flanking regions. We identified 105 polymorphisms, 50 of which were novel, including 8 nonsynonymous single-nucleotide polymorphisms, 7 were novel. Expression constructs were created for the nonsynonymous single-nucleotide polymorphisms. HEK293 cells were transfected with constructs for wild type and variant allozymes; and recombinant proteins were measured by quantitative Western blot analysis. The most significant change in NPR3 protein was observed for the Arg146 variant allozyme, with 20{\%} of wild-type protein, primarily because of autophagy-dependent degradation. NPR3 structural modeling confirmed that the Arg146 variant protein was not compatible with wild-type conformation and could result in protein misfolding or instability. Conclusions-Multiple novel NPR3 genetic polymorphisms were identified in 3 ethnic groups. The Arg146 allozyme displayed a significant decrease in protein quantity because of degradation mediated predominantly by autophagy. This genetic variation could have a significant effect on the metabolism of natriuretic peptides with potential clinical implications.",
keywords = "Natriuretic peptide receptor-3, Natriuretic peptides, Pharmacogenetics, Polymorphism",
author = "Pereira, {Naveen Luke} and Dong Lin and Linda Pelleymounter and Irene Moon and Gail Stilling and Eckloff, {Bruce W.} and Wieben, {Eric D} and Redfield, {Margaret May} and Burnett, {John C Jr.} and Yee, {Vivien C.} and Weinshilboum, {Richard M}",
year = "2013",
month = "4",
doi = "10.1161/CIRCGENETICS.112.964742",
language = "English (US)",
volume = "6",
pages = "201--210",
journal = "Circulation. Genomic and precision medicine",
issn = "1942-325X",
publisher = "Lippincott Williams and Wilkins Ltd.",
number = "2",

}

TY - JOUR

T1 - Natriuretic peptide receptor-3 gene (NPR3)

T2 - Nonsynonymous polymorphism results in significant reduction in protein expression because of accelerated degradation

AU - Pereira, Naveen Luke

AU - Lin, Dong

AU - Pelleymounter, Linda

AU - Moon, Irene

AU - Stilling, Gail

AU - Eckloff, Bruce W.

AU - Wieben, Eric D

AU - Redfield, Margaret May

AU - Burnett, John C Jr.

AU - Yee, Vivien C.

AU - Weinshilboum, Richard M

PY - 2013/4

Y1 - 2013/4

N2 - Background-The primary role of natriuretic peptide receptor-3 (NPR3) or NPR-C is in the clearance of natriuretic peptides that play an important role in modulating intravascular volume and vascular tone. Genetic variation in NPR3 has been associated with variation in blood pressure and obesity. Despite the importance of NPR3, sequence variation in the gene has not been addressed using DNA from different ethnic populations. We set out to identify and functionally characterize genetic variation in NPR3 in 3 ethnic groups. Methods and Results-DNA samples from 96 European American, 96 African American, and 96 Han Chinese American healthy subjects were used to resequence NPR3 exons, splice junctions, and flanking regions. We identified 105 polymorphisms, 50 of which were novel, including 8 nonsynonymous single-nucleotide polymorphisms, 7 were novel. Expression constructs were created for the nonsynonymous single-nucleotide polymorphisms. HEK293 cells were transfected with constructs for wild type and variant allozymes; and recombinant proteins were measured by quantitative Western blot analysis. The most significant change in NPR3 protein was observed for the Arg146 variant allozyme, with 20% of wild-type protein, primarily because of autophagy-dependent degradation. NPR3 structural modeling confirmed that the Arg146 variant protein was not compatible with wild-type conformation and could result in protein misfolding or instability. Conclusions-Multiple novel NPR3 genetic polymorphisms were identified in 3 ethnic groups. The Arg146 allozyme displayed a significant decrease in protein quantity because of degradation mediated predominantly by autophagy. This genetic variation could have a significant effect on the metabolism of natriuretic peptides with potential clinical implications.

AB - Background-The primary role of natriuretic peptide receptor-3 (NPR3) or NPR-C is in the clearance of natriuretic peptides that play an important role in modulating intravascular volume and vascular tone. Genetic variation in NPR3 has been associated with variation in blood pressure and obesity. Despite the importance of NPR3, sequence variation in the gene has not been addressed using DNA from different ethnic populations. We set out to identify and functionally characterize genetic variation in NPR3 in 3 ethnic groups. Methods and Results-DNA samples from 96 European American, 96 African American, and 96 Han Chinese American healthy subjects were used to resequence NPR3 exons, splice junctions, and flanking regions. We identified 105 polymorphisms, 50 of which were novel, including 8 nonsynonymous single-nucleotide polymorphisms, 7 were novel. Expression constructs were created for the nonsynonymous single-nucleotide polymorphisms. HEK293 cells were transfected with constructs for wild type and variant allozymes; and recombinant proteins were measured by quantitative Western blot analysis. The most significant change in NPR3 protein was observed for the Arg146 variant allozyme, with 20% of wild-type protein, primarily because of autophagy-dependent degradation. NPR3 structural modeling confirmed that the Arg146 variant protein was not compatible with wild-type conformation and could result in protein misfolding or instability. Conclusions-Multiple novel NPR3 genetic polymorphisms were identified in 3 ethnic groups. The Arg146 allozyme displayed a significant decrease in protein quantity because of degradation mediated predominantly by autophagy. This genetic variation could have a significant effect on the metabolism of natriuretic peptides with potential clinical implications.

KW - Natriuretic peptide receptor-3

KW - Natriuretic peptides

KW - Pharmacogenetics

KW - Polymorphism

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U2 - 10.1161/CIRCGENETICS.112.964742

DO - 10.1161/CIRCGENETICS.112.964742

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JO - Circulation. Genomic and precision medicine

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