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Diagnosis of N ‐acetylglutamate synthase deficiency by use of cultured fibroblasts and avoidance of nonsense‐mediated mRNA decay
Author(s) -
Häberle J.,
Denecke J.,
Schmidt E.,
Koch H. G.
Publication year - 2003
Publication title -
journal of inherited metabolic disease
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 102
eISSN - 1573-2665
pISSN - 0141-8955
DOI - 10.1023/a:1025912417548
Subject(s) - nonsense mediated decay , cycloheximide , nonsense mutation , messenger rna , rna splicing , biology , microbiology and biotechnology , gene , stop codon , allele , nonsense , translation (biology) , mutation , genetics , enzyme , protein biosynthesis , rna , biochemistry , missense mutation
Molecular diagnosis of N ‐acetylglutamate synthase deficiency (NAGSD) has become possible now that the corresponding gene has been identified. We describe the genetic analysis of a patient with NAGSD using low‐level transcripts derived from cultured fibroblasts. One defective allele (c.1306–1307insT) was detected by PCR amplification. However, the transcript from a second mutation (IVS3‐2A>T), causing aberrant splicing with the generation of a premature termination codon, was not detected until interference of nonsense‐mediated mRNA decay was abrogated by the translation inhibitor cycloheximide. We demonstrate that low‐level transcripts in cells that do not express significant enzyme activity are a valuable tool for molecular studies of genetic alterations, and suggest routine abrogation of nonsense‐mediated mRNA decay using cycloheximide when transcript analysis is performed.