Evidence for degradation of mRNA encoding alpha-L-iduronidase in Hurler fibroblasts with premature termination alleles.
Mutations in the gene encoding alpha-L-iduronidase (IDUA) are the cause of Hurler syndrome. Fibroblasts from patients homozygous for nonsense IDUA alleles have much reduced mRNA detectable by Northern analysis, as has been observed in many other instances of premature translation termination. Yet RT-PCR (reverse transcription followed by PCR amplification) showed a normal level of a segment covering exons 1 and 2 in Hurler cells homozygous for alleles bearing the nonsense mutations, Q70X or W402X. The 3' end of the segment was between exons 2 and 4. The results indicate that the nonsense RNA was degraded to fragment(s), independent of the position of the mutation (exon 2 or exon 9, respectively). Treatment of the cells with cycloheximide resulted in some increase of intact mRNA, suggesting that translation is required for mRNA degradation.
Menon
K P
KP
Department of Biological Chemistry, UCLA School of Medicine 90024-1737.
Neufeld
E F
EF
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D017931:DNA Primers / Q000235:genetics; D005091:Exons; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007068:Iduronidase / Q000235:genetics*; D008969:Molecular Sequence Data; D008059:Mucopolysaccharidosis I / Q000235:genetics* / Q000378:metabolism*; D009154:Mutation; D010443:Peptide Chain Termination, Translational / Q000235:genetics; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*
D017931:DNA Primers; D012333:RNA, Messenger; D003513:Cycloheximide; D007068:Iduronidase