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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
7
|
| pubmed:dateCreated |
1992-9-3
|
| pubmed:abstractText |
This study compares the synthesis of mutant type I collagen in cultured dermal fibroblasts and trabecular osteoblasts that were isolated from a patient with moderately severe osteogenesis imperfecta (type IV). Previous study of this patient's dermal fibroblasts revealed a 2000 dalton deletion located in cyanogen bromide peptide 4 of alpha 2(I)-collagen. The phenotype of the bone cell cultures was defined by a 3-4 day logarithmic phase doubling time, predominantly type I collagen production over type III and alkaline phosphatase activity 13.5 times dermal fibroblast levels. The current study revealed that both fibroblasts and osteoblasts synthesized a normal and a shortened alpha 2(I) chain, each as the product of separate alleles. Following pepsin treatment of the procollagens, a shortened alpha 1(I) chain was also seen in both cell types. Cyanogen bromide peptide mapping of osteoblast alpha-chains demonstrated the same deletions in the cyanogen bromide peptide 4 as observed in the fibroblast cyanogen bromide maps. PAGE analysis of oligonucleotide-specific cDNA that was reverse transcribed from RNA isolated from fibroblasts and osteoblasts also demonstrated the presence of two bands, one the normal size of alpha 2(I) cDNA and a second species that was smaller by 54 base pairs. Sequencing of polymerase chain reaction-amplified cDNA fragments revealed an in-frame deletion of exon 12. This finding was confirmed by the RNase protection method. Genomic DNA sequencing detected a T----G point mutation in the second position of the 5' splice donor site of intron 12. Therefore, in this patient with osteogenesis imperfecta there was no qualitative alteration in the osteoblast-specific expression of this mutant alpha 2(I)-collagen allele compared to dermal fibroblasts.
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Jul
|
| pubmed:issn |
0884-0431
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
7
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
793-805
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading |
pubmed-meshheading:1642148-Alkaline Phosphatase,
pubmed-meshheading:1642148-Amino Acid Sequence,
pubmed-meshheading:1642148-Base Sequence,
pubmed-meshheading:1642148-Cells, Cultured,
pubmed-meshheading:1642148-Collagen,
pubmed-meshheading:1642148-DNA,
pubmed-meshheading:1642148-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:1642148-Exons,
pubmed-meshheading:1642148-Female,
pubmed-meshheading:1642148-Fibroblasts,
pubmed-meshheading:1642148-Humans,
pubmed-meshheading:1642148-Middle Aged,
pubmed-meshheading:1642148-Molecular Sequence Data,
pubmed-meshheading:1642148-Mutation,
pubmed-meshheading:1642148-Osteoblasts,
pubmed-meshheading:1642148-Osteogenesis Imperfecta,
pubmed-meshheading:1642148-Polymerase Chain Reaction,
pubmed-meshheading:1642148-Procollagen,
pubmed-meshheading:1642148-Transcription, Genetic
|
| pubmed:year |
1992
|
| pubmed:articleTitle |
Expression of mutant alpha (I)-procollagen in osteoblast and fibroblast cultures from a proband with osteogenesis imperfecta type IV.
|
| pubmed:affiliation |
Division of Geriatric Medicine and Gerontology, Johns Hopkins University School of Medicine, Baltimore, Maryland.
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
|