Abstract
Skin fibroblasts from a proband with mild osteogenesis imperfecta (type I) synthesized normal proα2(I) chains and shortened proα2(I) chains of type-I procollagen. The type-I collagen that contained the shortened α2(I) chains was thermally unstable in that it was cleaved at 30°C by a mixture of trypsin and chymotrypsin. The mutation generating the shortened proα2(I) chains was shown to be a deletion of 19 base pairs from + 4 to + 22 of intron 13 of the COL1A2 gene by sequencing of genomic DNA and allele-specific oligonucleotide hybridization. The same mutation was found in the proband's affected father. Probe-protection experiments with S1 nuclease demonstrated that about 88% of the RNA transcripts from the mutated allele were spliced by exon skipping from exon 12 to exon 14 and that about 12% of the RNA transcripts were normally spliced. There was no evidence for use of cryptic splice sites, even though two cryptic splice sites had more favorable statistical scores and ΔG∘37 values than the new site that was created by the mutation and that was used for splicing of 12% of the transcripts into a normal mRNA. Comparison of the results with observations on 17 previously reported mutations that produced in-frame deletions of amino acids from the triple-helical domain of type-I collagen indicated that deletions in the N-terminal half of the α2(I) chain tended to produce milder phenotypes than similar deletions elsewhere in the α1(I) or α2(I) chains.
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Bonadio J, Ramirez F, Barr M (1990) An intron mutation in the human α1(I) collagen gene alters the efficiency of pre-mRNA splicing and is associated with osteogenesis imperfecta type II. J Biol Chem 265:2262–2268
Byers PH, Wallis GA, Willing MC (1991) Osteogenesis imperfecta: translation of mutation to phenotype. J Med Genet 28:433–442
Constantinou CD, Vogel BE, Jeffrey JJ, Prockop DJ (1987) The A and B fragments of normal type I procollagen have a similar thermal stability to proteinase digestion but are selectively destabilized by structural mutations. Eur J Biochem 163:247–251
Constantinou CD, Nielsen KB, Prockop DJ (1989) A lethal variant of osteogenesis imperfecta has a single base mutation that substitutes cysteine for glycine 904 of the α1(I) chain of type I procollagen. The asymptomatic mother has an unidentified mutation producing an overmodified and unstable type I procollagen. J Clin Invest 83:574–584
Engel J, Prockop DJ (1991) The zipper-like folding of collagen triple helices and the effects of mutations that disrupt the zipper. Annu Rev Biophys Biophys Chem 20:137–152
Freier S, Kierzek R, Jaeger JA, Sugimoto N, Caruthers MH, Neilson T, Turner DH (1986) Improved free-energy parameters for predictions of RNA. Proc Natl Acad Sci USA 83:9373–9377
Hawkins JR, Superti-Furga A, Steinmann B, Dalgleish R (1991) A 9-base pair deletion in COL1A1 in a lethal variant of osteogenesis imperfecta. J Biol Chem 266:22370–22374
Jacob M, Gallinaro H (1989) The 5′ splice site: phylogenetic evolution and variable geometry of association with U1RNA. Nucleic Acids Res 17:2159–2180
Kuivaniemi H, Tromp, G, Chu M-L, Prockop DJ (1988) Structure of a full-length cDNA clone for the preproα2(I) chain of human type I procollagen. Biochem J 252:633–640
Kuivaniemi H, Kontusaari S, Tromp G, Zhao M, Sabol C, Prockop DJ (1990) Identical G + 1 to A mutations in three different introns of the type III procollagen gene (COL3A1) produce different patterns of RNA splicing in three variants of Ehlers-Danlos syndrome IV. J Biol Chem 265:12067–12074
Kuivaniemi H, Tromp G, Prockop DJ (1991) Mutations in collagen genes: causes of rare and some common diseases in humans. FASEB J 5:2052–2050
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Nakajima H, Tada S (1971) Multiple osteogenesis imperfecta in one family. Seikei Geka (Jpn J Orthop Surg) 22:733–738
Pihlajaniemi T, Dickson LA, Pope FM, Lorhone VR, Nicholls A, Prockop DJ, Myers JC (1984) Osteogenesis imperfecta: cloning of a pro-α2(I) collagen gene with a frameshift mutation. J Biol Chem 259:12941–12944
Prockop DJ (1990) Mutations that alter the primary structure of type I collagen. J Biol Chem 265:15349–15352
Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N (1985) Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230:1350–1354
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Shapiro MB, Senapathy P (1987) RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression. Nucleic Acids Res 15:7155–7174
Spotila LD, Constantinou CD, Sereda L, Ganguly A, Riggs BL, Prockop DJ (1991) Mutation in a gene for type I procollagen (COL1A2) in a woman with postmenopausal osteoporosis: evidence for phenotypic and genotypic overlap with mild osteogenesis imperfecta. Proc Natl Acad Sci USA 88:5423–5427
Studencki AB, Wallace RB (1984) Allele-specific hybridization using oligonucleotide probes of very high specific activity: discrimination of the human β A- and β s-globin genes. DNA 3:7–15
Sykes B, Ogilvie D, Wordsworth P, Wallis G, Mathew C, Beighton P, Nicholls A, Pope FM, Thompson E, Tsipouras P, Schwartz R, Jensson O, Arnason A, Boresen A-L, Heiberg A, Frey D, Steinmann B (1990) Consistent linkage of dominantly inherited osteogenesis imperfecta to the type I collagen loci: COL1A1 and COL1A2. Am J Hum Genet 46:293–307
Tromp G, Prockop DJ (1988) Single base mutation in the proα2(I) collagen gene that causes efficient splicing of RNA from exon 27 to exon 29 and synthesis of a shortened but in-frame proα2(I) chain. Proc Natl Acad Sci USA 85:5254–5258
Wet WJ de, Pihlajaniemi T, Myers J, Kelly TE, Prockop DJ (1983) Synthesis of a shortened pro-α2(I) chain and decreased synthesis of pro-α2(I) chains in a proband with osteogenesis imperfecta. J Biol Chem 258:7721–7728
Wet WJ de, Bernard M, Benson-Chanda V, Chu M-L, Dickson L, Weil D, Ramirez F (1987) Organization of the human pro-α2(I) collagen gene. J Biol Chem 262:16032–16036
Zorn AM, Krieg PA (1991) PCR analysis of alternative splicing pathways: identification of artifacts generated by heteroduplex formation. Biotechniques 11:180–184
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Zhuang, J., Tromp, G., Kuivaniemi, H. et al. Deletion of 19 base pairs in intron 13 of the gene for the proα2(I) chain of type-I procollagen (COL1A2) causes exon skipping in a proband with type-I osteogenesis imperfecta. Hum Genet 91, 210–216 (1993). https://doi.org/10.1007/BF00218258
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DOI: https://doi.org/10.1007/BF00218258