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 G.
AU - Cutting, Garry R.
PY - 1991
Y1 - 1991
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
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U2 - 10.1172/JCI115510
DO - 10.1172/JCI115510
M3 - Article
C2 - 1721624
AN - SCOPUS:0026322140
SN - 0021-9738
VL - 88
SP - 1880
EP - 1885
JO - Journal of Clinical Investigation
JF - Journal of Clinical Investigation
IS - 6
ER -