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Am J Hum Genet. 1998 Jan; 62(1): 98–110.
PMCID: PMC1376813

Analysis of the COL1A1 and COL1A2 genes by PCR amplification and scanning by conformation-sensitive gel electrophoresis identifies only COL1A1 mutations in 15 patients with osteogenesis imperfecta type I: identification of common sequences of null-allele mutations.


Although >90% of patients with osteogenesis imperfecta (OI) have been estimated to have mutations in the COL1A1 and COL1A2 genes for type I procollagen, mutations have been difficult to detect in all patients with the mildest forms of the disease (i.e., type I). In this study, we first searched for mutations in type I procollagen by analyses of protein and mRNA in fibroblasts from 10 patients with mild OI; no evidence of a mutation was found in 2 of the patients by the protein analyses, and no evidence of a mutation was found in 5 of the patients by the RNA analyses. We then searched for mutations in the original 10 patients and in 5 additional patients with mild OI, by analysis of genomic DNA. To assay the genomic DNA, we established a consensus sequence for the first 12 kb of the COL1A1 gene and for 30 kb of new sequences of the 38-kb COL1A2 gene. The sequences were then used to develop primers for PCR for the 103 exons and exon boundaries of the two genes. The PCR products were first scanned for heteroduplexes by conformation-sensitive gel electrophoresis, and then products containing heteroduplexes were sequenced. The results detected disease-causing mutations in 13 of the 15 patients and detected two additional probable disease-causing mutations in the remaining 2 patients. Analysis of the data developed in this study and elsewhere revealed common sequences for mutations causing null alleles.

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Selected References

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  • Ahmad NN, Dimascio J, Knowlton RG, Tasman WS. Stickler syndrome. A mutation in the nonhelical 3' end of type II procollagen gene. Arch Ophthalmol. 1995 Nov;113(11):1454–1457. [PubMed]
  • Ala-Kokko L, Kvist AP, Metsäranta M, Kivirikko KI, de Crombrugghe B, Prockop DJ, Vuorio E. Conservation of the sizes of 53 introns and over 100 intronic sequences for the binding of common transcription factors in the human and mouse genes for type II procollagen (COL2A1). Biochem J. 1995 Jun 15;308(Pt 3):923–929. [PMC free article] [PubMed]
  • Bernard MP, Myers JC, Chu ML, Ramirez F, Eikenberry EF, Prockop DJ. Structure of a cDNA for the pro alpha 2 chain of human type I procollagen. Comparison with chick cDNA for pro alpha 2(I) identifies structurally conserved features of the protein and the gene. Biochemistry. 1983 Mar 1;22(5):1139–1145. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Boedtker H, Finer M, Aho S. The structure of the chicken alpha 2 collagen gene. Ann N Y Acad Sci. 1985;460:85–116. [PubMed]
  • Brown DM, Vandenburgh K, Kimura AE, Weingeist TA, Sheffield VC, Stone EM. Novel frameshift mutations in the procollagen 2 gene (COL2A1) associated with Stickler syndrome (hereditary arthro-ophthalmopathy). Hum Mol Genet. 1995 Jan;4(1):141–142. [PubMed]
  • Chipman SD, Sweet HO, McBride DJ, Jr, Davisson MT, Marks SC, Jr, Shuldiner AR, Wenstrup RJ, Rowe DW, Shapiro JR. Defective pro alpha 2(I) collagen synthesis in a recessive mutation in mice: a model of human osteogenesis imperfecta. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1701–1705. [PMC free article] [PubMed]
  • Chu ML, de Wet W, Bernard M, Ramirez F. Fine structural analysis of the human pro-alpha 1 (I) collagen gene. Promoter structure, AluI repeats, and polymorphic transcripts. J Biol Chem. 1985 Feb 25;260(4):2315–2320. [PubMed]
  • D'Alessio M, Bernard M, Pretorius PJ, de Wet W, Ramirez F, Pretorious PJ. Complete nucleotide sequence of the region encompassing the first twenty-five exons of the human pro alpha 1(I) collagen gene (COL1A1) Gene. 1988 Jul 15;67(1):105–115. [PubMed]
  • Dalgleish R. The human type I collagen mutation database. Nucleic Acids Res. 1997 Jan 1;25(1):181–187. [PMC free article] [PubMed]
  • de Wet W, Bernard M, Benson-Chanda V, Chu ML, Dickson L, Weil D, Ramirez F. Organization of the human pro-alpha 2(I) collagen gene. J Biol Chem. 1987 Nov 25;262(33):16032–16036. [PubMed]
  • Dickson LA, de Wet W, Di Liberto M, Weil D, Ramirez F. Analysis of the promoter region and the N-propeptide domain of the human pro alpha 2(I) collagen gene. Nucleic Acids Res. 1985 May 24;13(10):3427–3438. [PMC free article] [PubMed]
  • Dickson LA, Pihlajaniemi T, Deak S, Pope FM, Nicholls A, Prockop DJ, Myers JC. Nuclease S1 mapping of a homozygous mutation in the carboxyl-propeptide-coding region of the pro alpha 2(I) collagen gene in a patient with osteogenesis imperfecta. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4524–4528. [PMC free article] [PubMed]
  • Ganguly A, Baldwin CT, Strobel D, Conway D, Horton W, Prockop DJ. Heterozygous mutation in the G+5 position of intron 33 of the pro-alpha 2(I) gene (COL1A2) that causes aberrant RNA splicing and lethal osteogenesis imperfecta. Use of carbodiimide methods that decrease the extent of DNA sequencing necessary to define an unusual mutation. J Biol Chem. 1991 Jun 25;266(18):12035–12040. [PubMed]
  • Ganguly A, Prockop DJ. Detection of mismatched bases in double stranded DNA by gel electrophoresis. Electrophoresis. 1995 Oct;16(10):1830–1835. [PubMed]
  • Ganguly A, Rock MJ, Prockop DJ. Conformation-sensitive gel electrophoresis for rapid detection of single-base differences in double-stranded PCR products and DNA fragments: evidence for solvent-induced bends in DNA heteroduplexes. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10325–10329. [PMC free article] [PubMed]
  • Hanke M, Wink M. Direct DNA sequencing of PCR-amplified vector inserts following enzymatic degradation of primer and dNTPs. Biotechniques. 1994 Nov;17(5):858–860. [PubMed]
  • Jego N, Thomas G, Hamelin R. Short direct repeats flanking deletions, and duplicating insertions in p53 gene in human cancers. Oncogene. 1993 Jan;8(1):209–213. [PubMed]
  • Körkkö J, Kuivaniemi H, Paassilta P, Zhuang J, Tromp G, DePaepe A, Prockop DJ, Ala-Kokko L. Two new recurrent nucleotide mutations in the COL1A1 gene in four patients with osteogenesis imperfecta: about one-fifth are recurrent. Hum Mutat. 1997;9(2):148–156. [PubMed]
  • Kuivaniemi H, Tromp G, Chu ML, Prockop DJ. Structure of a full-length cDNA clone for the prepro alpha 2(I) chain of human type I procollagen. Comparison with the chicken gene confirms unusual patterns of gene conservation. Biochem J. 1988 Jun 15;252(3):633–640. [PMC free article] [PubMed]
  • Kuivaniemi H, Tromp G, Prockop DJ. Mutations in fibrillar collagens (types I, II, III, and XI), fibril-associated collagen (type IX), and network-forming collagen (type X) cause a spectrum of diseases of bone, cartilage, and blood vessels. Hum Mutat. 1997;9(4):300–315. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Mättä A, Bornstein P, Penttinen RP. Highly conserved sequences in the 3'-untranslated region of the COL1A1 gene bind cell-specific nuclear proteins. FEBS Lett. 1991 Feb 11;279(1):9–13. [PubMed]
  • Myers JC, Dickson LA, de Wet WJ, Bernard MP, Chu ML, Di Liberto M, Pepe G, Sangiorgi FO, Ramirez F. Analysis of the 3' end of the human pro-alpha 2(I) collagen gene. Utilization of multiple polyadenylation sites in cultured fibroblasts. J Biol Chem. 1983 Aug 25;258(16):10128–10135. [PubMed]
  • Nakai K, Sakamoto H. Construction of a novel database containing aberrant splicing mutations of mammalian genes. Gene. 1994 Apr 20;141(2):171–177. [PubMed]
  • Ollila J, Lappalainen I, Vihinen M. Sequence specificity in CpG mutation hotspots. FEBS Lett. 1996 Nov 4;396(2-3):119–122. [PubMed]
  • Prockop DJ. Mutations that alter the primary structure of type I collagen. The perils of a system for generating large structures by the principle of nucleated growth. J Biol Chem. 1990 Sep 15;265(26):15349–15352. [PubMed]
  • Prockop DJ, Kivirikko KI. Collagens: molecular biology, diseases, and potentials for therapy. Annu Rev Biochem. 1995;64:403–434. [PubMed]
  • Redford-Badwal DA, Stover ML, Valli M, McKinstry MB, Rowe DW. Nuclear retention of COL1A1 messenger RNA identifies null alleles causing mild osteogenesis imperfecta. J Clin Invest. 1996 Feb 15;97(4):1035–1040. [PMC free article] [PubMed]
  • Saban J, King D. PCR genotyping of oim mutant mice. Biotechniques. 1996 Aug;21(2):190–192. [PubMed]
  • Sherwood AL, Bottenus RE, Martzen MR, Bornstein P. Structural and functional analysis of the first intron of the human alpha 2(I) collagen-encoding gene. Gene. 1990 May 14;89(2):239–244. [PubMed]
  • Sokolov BP, Prytkov AN, Tromp G, Knowlton RG, Prockop DJ. Exclusion of COL1A1, COL1A2, and COL3A1 genes as candidate genes for Ehlers-Danlos syndrome type I in one large family. Hum Genet. 1991 Dec;88(2):125–129. [PubMed]
  • Sykes B, Ogilvie D, Wordsworth P, Wallis G, Mathew C, Beighton P, Nicholls A, Pope FM, Thompson E, Tsipouras P, et al. Consistent linkage of dominantly inherited osteogenesis imperfecta to the type I collagen loci: COL1A1 and COL1A2. Am J Hum Genet. 1990 Feb;46(2):293–307. [PMC free article] [PubMed]
  • Tromp G, Prockop DJ. Single base mutation in the pro alpha 2(I) collagen gene that causes efficient splicing of RNA from exon 27 to exon 29 and synthesis of a shortened but in-frame pro alpha 2(I) chain. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5254–5258. [PMC free article] [PubMed]
  • Vasan NS, Kuivaniemi H, Vogel BE, Minor RR, Wootton JA, Tromp G, Weksberg R, Prockop DJ. A mutation in the pro alpha 2(I) gene (COL1A2) for type I procollagen in Ehlers-Danlos syndrome type VII: evidence suggesting that skipping of exon 6 in RNA splicing may be a common cause of the phenotype. Am J Hum Genet. 1991 Feb;48(2):305–317. [PMC free article] [PubMed]
  • Werle E, Schneider C, Renner M, Völker M, Fiehn W. Convenient single-step, one tube purification of PCR products for direct sequencing. Nucleic Acids Res. 1994 Oct 11;22(20):4354–4355. [PMC free article] [PubMed]
  • Westerhausen A, Constantinou CD, Pack M, Peng MZ, Hanning C, Olsen AS, Prockop DJ. Completion of the last half of the structure of the human gene for the Pro alpha 1 (I) chain of type I procollagen (COL1A1). Matrix. 1991 Dec;11(6):375–379. [PubMed]
  • Willing MC, Deschenes SP, Scott DA, Byers PH, Slayton RL, Pitts SH, Arikat H, Roberts EJ. Osteogenesis imperfecta type I: molecular heterogeneity for COL1A1 null alleles of type I collagen. Am J Hum Genet. 1994 Oct;55(4):638–647. [PMC free article] [PubMed]
  • Willing MC, Deschenes SP, Slayton RL, Roberts EJ. Premature chain termination is a unifying mechanism for COL1A1 null alleles in osteogenesis imperfecta type I cell strains. Am J Hum Genet. 1996 Oct;59(4):799–809. [PMC free article] [PubMed]
  • Willing MC, Pruchno CJ, Atkinson M, Byers PH. Osteogenesis imperfecta type I is commonly due to a COL1A1 null allele of type I collagen. Am J Hum Genet. 1992 Sep;51(3):508–515. [PMC free article] [PubMed]

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