Severe deficiency of cystic fibrosis transmembrane conductance regulator messenger RNA carrying nonsense mutations R553X and W1316X in respiratory epithelial cells of patients with cystic fibrosis

Ada Hamosh, Bruce C. Trapnell, Pamela L. Zeitlin, Chahrzad Montrose-Rafizadeh, Beryl J. Rosenstein, Ronald Crystal, Garry R. Cutting

Research output: Contribution to journalArticle

114 Citations (Scopus)

Abstract

Cystic fibrosis (CF) is the most common, lethal inherited disorder in the Caucasian population. We have recently reported two African-American patients with nonsense mutations in each CF gene and severe pancreatic disease, but mild pulmonary disease. In order to examine the effect of these nonsense mutations on CF gene expression, bronchial and nasal epithelial cells were obtained from one of these patients (no. 246), a compound heterozygote for nonsense mutations R553X and W1316X; a healthy normal individual; a patient (no. 528) homozygous for the common CF mutation (ΔF508); and a CF patient (no. 272) who carries the R553X mutation and a mis-sense mutation, S549N. When mRNA from bronchial cells of the normal individual, the ΔF508 homozygote, and the S549N/R553X compound heterozygote was reverse transcribed and amplified by polymerase chain reaction using primers derived from the CF gene, DNA fragments of the predicted size were observed. However, patient no. 246 with nonsense mutations in each CF gene has no detectable cystic fibrosis transmembrane conductance regulator (CFTR) messenger RNA, and therefore should have severely diminished, and possibly absent, CFTR protein. Furthermore, < 2% of the CFTR transcripts in nasal epithelial cells from patient no. 272 (S549N/R553X) were derived from the gene with the nonsense mutation. We conclude that severe reduction in CFTR mRNA causes CF, but can have different consequences in the lung and pancreas.

Original languageEnglish
Pages (from-to)1880-1885
Number of pages6
JournalJournal of Clinical Investigation
Volume88
Issue number6
DOIs
Publication statusPublished - 1 Jan 1991
Externally publishedYes

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Nonsense Codon
Cystic Fibrosis
Epithelial Cells
Messenger RNA
Heterozygote
Nose
Mutation
Genes
Pancreatic Diseases
Homozygote
African Americans
Lung Diseases
Pancreas
Gene Expression
Polymerase Chain Reaction
Lung
DNA

Keywords

  • Allele-specific oligonucleotides
  • Genotype
  • Phenotype
  • Polymerase chain reaction
  • Reverse transcription

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Severe deficiency of cystic fibrosis transmembrane conductance regulator messenger RNA carrying nonsense mutations R553X and W1316X in respiratory epithelial cells of patients with cystic fibrosis. / Hamosh, Ada; Trapnell, Bruce C.; Zeitlin, Pamela L.; Montrose-Rafizadeh, Chahrzad; Rosenstein, Beryl J.; Crystal, Ronald; Cutting, Garry R.

In: Journal of Clinical Investigation, Vol. 88, No. 6, 01.01.1991, p. 1880-1885.

Research output: Contribution to journalArticle

Hamosh, Ada ; Trapnell, Bruce C. ; Zeitlin, Pamela L. ; Montrose-Rafizadeh, Chahrzad ; Rosenstein, Beryl J. ; Crystal, Ronald ; Cutting, Garry R. / Severe deficiency of cystic fibrosis transmembrane conductance regulator messenger RNA carrying nonsense mutations R553X and W1316X in respiratory epithelial cells of patients with cystic fibrosis. In: Journal of Clinical Investigation. 1991 ; Vol. 88, No. 6. pp. 1880-1885.
@article{6c37029793f84c5a8a045fed8900dcc3,
title = "Severe deficiency of cystic fibrosis transmembrane conductance regulator messenger RNA carrying nonsense mutations R553X and W1316X in respiratory epithelial cells of patients with cystic fibrosis",
abstract = "Cystic fibrosis (CF) is the most common, lethal inherited disorder in the Caucasian population. We have recently reported two African-American patients with nonsense mutations in each CF gene and severe pancreatic disease, but mild pulmonary disease. In order to examine the effect of these nonsense mutations on CF gene expression, bronchial and nasal epithelial cells were obtained from one of these patients (no. 246), a compound heterozygote for nonsense mutations R553X and W1316X; a healthy normal individual; a patient (no. 528) homozygous for the common CF mutation (ΔF508); and a CF patient (no. 272) who carries the R553X mutation and a mis-sense mutation, S549N. When mRNA from bronchial cells of the normal individual, the ΔF508 homozygote, and the S549N/R553X compound heterozygote was reverse transcribed and amplified by polymerase chain reaction using primers derived from the CF gene, DNA fragments of the predicted size were observed. However, patient no. 246 with nonsense mutations in each CF gene has no detectable cystic fibrosis transmembrane conductance regulator (CFTR) messenger RNA, and therefore should have severely diminished, and possibly absent, CFTR protein. Furthermore, < 2{\%} of the CFTR transcripts in nasal epithelial cells from patient no. 272 (S549N/R553X) were derived from the gene with the nonsense mutation. We conclude that severe reduction in CFTR mRNA causes CF, but can have different consequences in the lung and pancreas.",
keywords = "Allele-specific oligonucleotides, Genotype, Phenotype, Polymerase chain reaction, Reverse transcription",
author = "Ada Hamosh and Trapnell, {Bruce C.} and Zeitlin, {Pamela L.} and Chahrzad Montrose-Rafizadeh and Rosenstein, {Beryl J.} and Ronald Crystal and Cutting, {Garry R.}",
year = "1991",
month = "1",
day = "1",
doi = "10.1172/JCI115510",
language = "English",
volume = "88",
pages = "1880--1885",
journal = "Journal of Clinical Investigation",
issn = "0021-9738",
publisher = "The American Society for Clinical Investigation",
number = "6",

}

TY - JOUR

T1 - Severe deficiency of cystic fibrosis transmembrane conductance regulator messenger RNA carrying nonsense mutations R553X and W1316X in respiratory epithelial cells of patients with cystic fibrosis

AU - Hamosh, Ada

AU - Trapnell, Bruce C.

AU - Zeitlin, Pamela L.

AU - Montrose-Rafizadeh, Chahrzad

AU - Rosenstein, Beryl J.

AU - Crystal, Ronald

AU - Cutting, Garry R.

PY - 1991/1/1

Y1 - 1991/1/1

N2 - Cystic fibrosis (CF) is the most common, lethal inherited disorder in the Caucasian population. We have recently reported two African-American patients with nonsense mutations in each CF gene and severe pancreatic disease, but mild pulmonary disease. In order to examine the effect of these nonsense mutations on CF gene expression, bronchial and nasal epithelial cells were obtained from one of these patients (no. 246), a compound heterozygote for nonsense mutations R553X and W1316X; a healthy normal individual; a patient (no. 528) homozygous for the common CF mutation (ΔF508); and a CF patient (no. 272) who carries the R553X mutation and a mis-sense mutation, S549N. When mRNA from bronchial cells of the normal individual, the ΔF508 homozygote, and the S549N/R553X compound heterozygote was reverse transcribed and amplified by polymerase chain reaction using primers derived from the CF gene, DNA fragments of the predicted size were observed. However, patient no. 246 with nonsense mutations in each CF gene has no detectable cystic fibrosis transmembrane conductance regulator (CFTR) messenger RNA, and therefore should have severely diminished, and possibly absent, CFTR protein. Furthermore, < 2% of the CFTR transcripts in nasal epithelial cells from patient no. 272 (S549N/R553X) were derived from the gene with the nonsense mutation. We conclude that severe reduction in CFTR mRNA causes CF, but can have different consequences in the lung and pancreas.

AB - Cystic fibrosis (CF) is the most common, lethal inherited disorder in the Caucasian population. We have recently reported two African-American patients with nonsense mutations in each CF gene and severe pancreatic disease, but mild pulmonary disease. In order to examine the effect of these nonsense mutations on CF gene expression, bronchial and nasal epithelial cells were obtained from one of these patients (no. 246), a compound heterozygote for nonsense mutations R553X and W1316X; a healthy normal individual; a patient (no. 528) homozygous for the common CF mutation (ΔF508); and a CF patient (no. 272) who carries the R553X mutation and a mis-sense mutation, S549N. When mRNA from bronchial cells of the normal individual, the ΔF508 homozygote, and the S549N/R553X compound heterozygote was reverse transcribed and amplified by polymerase chain reaction using primers derived from the CF gene, DNA fragments of the predicted size were observed. However, patient no. 246 with nonsense mutations in each CF gene has no detectable cystic fibrosis transmembrane conductance regulator (CFTR) messenger RNA, and therefore should have severely diminished, and possibly absent, CFTR protein. Furthermore, < 2% of the CFTR transcripts in nasal epithelial cells from patient no. 272 (S549N/R553X) were derived from the gene with the nonsense mutation. We conclude that severe reduction in CFTR mRNA causes CF, but can have different consequences in the lung and pancreas.

KW - Allele-specific oligonucleotides

KW - Genotype

KW - Phenotype

KW - Polymerase chain reaction

KW - Reverse transcription

UR - http://www.scopus.com/inward/record.url?scp=0026322140&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026322140&partnerID=8YFLogxK

U2 - 10.1172/JCI115510

DO - 10.1172/JCI115510

M3 - Article

VL - 88

SP - 1880

EP - 1885

JO - Journal of Clinical Investigation

JF - Journal of Clinical Investigation

SN - 0021-9738

IS - 6

ER -