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Temperature sensitivity of aberrant RNA splicing with a mutation in theG +5 position of intron 37 of the gene for type III procollagen from a patient with Ehlers–Danlos syndrome type IV
Author(s) -
Wu Yuli,
Kuivaniemi Helena,
Tromp Gerard,
Strobel Deike,
Romanic Anne M.,
Prockop Darwin J.
Publication year - 1993
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.1380020106
Subject(s) - intron , biology , rna , exon , splice site mutation , rna splicing , microbiology and biotechnology , exon skipping , gene , procollagen peptidase , small nuclear rna , messenger rna , mutant , genetics , non coding rna
A single‐base mutation in intron 37 of the gene for type III procollagen (COL3A1) was found in a proband with the type IV variant of Ehlers—Danlos syndrome. Probe‐protection experiments with S1 nuclease and RNA from fibroblasts incubated at 37°C demonstrated that about 35% of the total mRNA or about 70% of the mRNA from mutated allele was spliced by exon skipping. The effects of the mutation were temperature‐sensitive in that the amount of RNA from the mutated allele that was spliced by exon skipping was 87.1 ± 7.7% at 31°C, 70.1 ± 6.5% at 37°C, and 85.4 ± 11.1% at 42°C. The effects of temperature on aberrant RNA splicing were, therefore, the reverse of those reported for four previous mutants incollagen genes. The increase in abnormal RNA splicing when the temperature was raised from 31° to 37°C seen with previously reported mutants suggested that RNA–RNA hybridization of U1snRNA to the 5′‐splice site in the substrate may be limiting in the processing of transcripts from the mutated alleles, since RNA–RNA hybridizations become less favorable at higher temperatures. The decrease in abnormal RNA splicing seen here when the temperature was raised from 31° to 37°C suggested that protein–RNA or protein–protein binding steps become rate limiting with the G +5 mutation in intron 37 of the COL3A1 gene. © 1993 Wiley‐Liss, Inc.