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. 1993 Oct;92(4):1994–2002. doi: 10.1172/JCI116794

Defective splicing of mRNA from one COL1A1 allele of type I collagen in nondeforming (type I) osteogenesis imperfecta.

M L Stover 1, D Primorac 1, S C Liu 1, M B McKinstry 1, D W Rowe 1
PMCID: PMC288367  PMID: 8408653

Abstract

Osteogenesis imperfecta (OI) type I is the mildest form of heritable bone fragility resulting from mutations within the COL1A1 gene. We studied fibroblasts established from a child with OI type I and demonstrated underproduction of alpha 1 (I) collagen chains and alpha 1 (I) mRNA. Indirect RNase protection suggested two species of alpha 1 (I) mRNA, one of which was not collinear with fully spliced alpha 1 (I) mRNA. The noncollinear population was confined to the nuclear compartment of the cell, and contained the entire sequence of intron 26 and a G-->A transition in the first position of the intron donor site. The G-->A transition was also identified in the genomic DNA. The retained intron contained an in-frame stop codon and introduced an out-of-frame insertion within the collagen mRNA producing stop codons downstream of the insertion. These changes probably account for the failure of the mutant RNA to appear in the cytoplasm. Unlike other splice site mutations within collagen mRNA that resulted in exon skipping and a truncated but inframe RNA transcript, this mutation did not result in production of a defective collagen pro alpha 1 (I) chain. Instead, the mild nature of the disease in this case reflects failure to process the defective mRNA and thus the absence of a protein product from the mutant allele.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barsh G. S., David K. E., Byers P. H. Type I osteogenesis imperfecta: a nonfunctional allele for pro alpha 1 (I) chains of type I procollagen. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3838–3842. doi: 10.1073/pnas.79.12.3838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baserga S. J., Benz E. J., Jr Nonsense mutations in the human beta-globin gene affect mRNA metabolism. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2056–2060. doi: 10.1073/pnas.85.7.2056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bateman J. F., Lamande S. R., Dahl H. H., Chan D., Mascara T., Cole W. G. A frameshift mutation results in a truncated nonfunctional carboxyl-terminal pro alpha 1(I) propeptide of type I collagen in osteogenesis imperfecta. J Biol Chem. 1989 Jul 5;264(19):10960–10964. [PubMed] [Google Scholar]
  4. Bateman J. F., Moeller I., Hannagan M., Chan D., Cole W. G. Characterization of three osteogenesis imperfecta collagen alpha 1(I) glycine to serine mutations demonstrating a position-dependent gradient of phenotypic severity. Biochem J. 1992 Nov 15;288(Pt 1):131–135. doi: 10.1042/bj2880131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bonadio J., Ramirez F., Barr M. An intron mutation in the human alpha 1(I) collagen gene alters the efficiency of pre-mRNA splicing and is associated with osteogenesis imperfecta type II. J Biol Chem. 1990 Feb 5;265(4):2262–2268. [PubMed] [Google Scholar]
  6. Byers P. H. Brittle bones--fragile molecules: disorders of collagen gene structure and expression. Trends Genet. 1990 Sep;6(9):293–300. doi: 10.1016/0168-9525(90)90235-x. [DOI] [PubMed] [Google Scholar]
  7. Byers P. H., Shapiro J. R., Rowe D. W., David K. E., Holbrook K. A. Abnormal alpha 2-chain in type I collagen from a patient with a form of osteogenesis imperfecta. J Clin Invest. 1983 Mar;71(3):689–697. doi: 10.1172/JCI110815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Byers P. H., Steiner R. D. Osteogenesis imperfecta. Annu Rev Med. 1992;43:269–282. doi: 10.1146/annurev.me.43.020192.001413. [DOI] [PubMed] [Google Scholar]
  9. Byers P. H., Wallis G. A., Willing M. C. Osteogenesis imperfecta: translation of mutation to phenotype. J Med Genet. 1991 Jul;28(7):433–442. doi: 10.1136/jmg.28.7.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cheng J., Fogel-Petrovic M., Maquat L. E. Translation to near the distal end of the penultimate exon is required for normal levels of spliced triosephosphate isomerase mRNA. Mol Cell Biol. 1990 Oct;10(10):5215–5225. doi: 10.1128/mcb.10.10.5215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chipman S. D., Shapiro J. R., McKinstry M. B., Stover M. L., Branson P., Rowe D. W. Expression of mutant alpha (I)-procollagen in osteoblast and fibroblast cultures from a proband with osteogenesis imperfecta type IV. J Bone Miner Res. 1992 Jul;7(7):793–805. doi: 10.1002/jbmr.5650070709. [DOI] [PubMed] [Google Scholar]
  12. Chipman S. D., Sweet H. O., McBride D. J., Jr, Davisson M. T., Marks S. C., Jr, Shuldiner A. R., Wenstrup R. J., Rowe D. W., Shapiro J. R. 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. doi: 10.1073/pnas.90.5.1701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chou Q., Russell M., Birch D. E., Raymond J., Bloch W. Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res. 1992 Apr 11;20(7):1717–1723. doi: 10.1093/nar/20.7.1717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Chu M. L., Myers J. C., Bernard M. P., Ding J. F., Ramirez F. Cloning and characterization of five overlapping cDNAs specific for the human pro alpha 1(I) collagen chain. Nucleic Acids Res. 1982 Oct 11;10(19):5925–5934. doi: 10.1093/nar/10.19.5925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Chu M. L., Rowe D., Nicholls A. C., Pope F. M., Prockop D. J. Presence of translatable mRNA for pro alpha 2(I) chains in fibroblasts from a patient with osteogenesis imperfecta whose type I collagen does not contain alpha 2(I) chains. Coll Relat Res. 1984 Oct;4(5):389–394. doi: 10.1016/s0174-173x(84)80006-0. [DOI] [PubMed] [Google Scholar]
  16. Chu M. L., Williams C. J., Pepe G., Hirsch J. L., Prockop D. J., Ramirez F. Internal deletion in a collagen gene in a perinatal lethal form of osteogenesis imperfecta. Nature. 1983 Jul 7;304(5921):78–80. doi: 10.1038/304078a0. [DOI] [PubMed] [Google Scholar]
  17. Dominski Z., Kole R. Cooperation of pre-mRNA sequence elements in splice site selection. Mol Cell Biol. 1992 May;12(5):2108–2114. doi: 10.1128/mcb.12.5.2108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  19. Genovese C., Brufsky A., Shapiro J., Rowe D. Detection of mutations in human type I collagen mRNA in osteogenesis imperfecta by indirect RNase protection. J Biol Chem. 1989 Jun 5;264(16):9632–9637. [PubMed] [Google Scholar]
  20. Genovese C., Rowe D. Analysis of cytoplasmic and nuclear messenger RNA in fibroblasts from patients with type I osteogenesis imperfecta. Methods Enzymol. 1987;145:223–235. doi: 10.1016/0076-6879(87)45012-x. [DOI] [PubMed] [Google Scholar]
  21. Genovese C., Rowe D., Kream B. Construction of DNA sequences complementary to rat alpha 1 and alpha 2 collagen mRNA and their use in studying the regulation of type I collagen synthesis by 1,25-dihydroxyvitamin D. Biochemistry. 1984 Dec 4;23(25):6210–6216. doi: 10.1021/bi00320a049. [DOI] [PubMed] [Google Scholar]
  22. Hamosh A., Trapnell B. C., Zeitlin P. L., Montrose-Rafizadeh C., Rosenstein B. J., Crystal R. G., Cutting G. R. Severe deficiency of cystic fibrosis transmembrane conductance regulator messenger RNA carrying nonsense mutations R553X and W1316X in respiratory epithelial cells of patients with cystic fibrosis. J Clin Invest. 1991 Dec;88(6):1880–1885. doi: 10.1172/JCI115510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hawkins J. D. A survey on intron and exon lengths. Nucleic Acids Res. 1988 Nov 11;16(21):9893–9908. doi: 10.1093/nar/16.21.9893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kadler K. E., Torre-Blanco A., Adachi E., Vogel B. E., Hojima Y., Prockop D. J. A type I collagen with substitution of a cysteine for glycine-748 in the alpha 1(I) chain copolymerizes with normal type I collagen and can generate fractallike structures. Biochemistry. 1991 May 21;30(20):5081–5088. doi: 10.1021/bi00234a035. [DOI] [PubMed] [Google Scholar]
  25. Koivisto U. M., Turtola H., Aalto-Setälä K., Top B., Frants R. R., Kovanen P. T., Syvänen A. C., Kontula K. The familial hypercholesterolemia (FH)-North Karelia mutation of the low density lipoprotein receptor gene deletes seven nucleotides of exon 6 and is a common cause of FH in Finland. J Clin Invest. 1992 Jul;90(1):219–228. doi: 10.1172/JCI115839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kuivaniemi H., Kontusaari S., Tromp G., Zhao M. J., Sabol C., Prockop D. J. 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. An explanation for exon skipping some mutations and not others. J Biol Chem. 1990 Jul 15;265(20):12067–12074. [PubMed] [Google Scholar]
  27. Kuivaniemi H., Sabol C., Tromp G., Sippola-Thiele M., Prockop D. J. A 19-base pair deletion in the pro-alpha 2(I) gene of type I procollagen that causes in-frame RNA splicing from exon 10 to exon 12 in a proband with atypical osteogenesis imperfecta and in his asymptomatic mother. J Biol Chem. 1988 Aug 15;263(23):11407–11413. [PubMed] [Google Scholar]
  28. Kuivaniemi H., Tromp G., Prockop D. J. Mutations in collagen genes: causes of rare and some common diseases in humans. FASEB J. 1991 Apr;5(7):2052–2060. doi: 10.1096/fasebj.5.7.2010058. [DOI] [PubMed] [Google Scholar]
  29. Lichtler A., Barrett N. L., Carmichael G. G. Simple, inexpensive preparation of T1/T2 ribonuclease suitable for use in RNase protection experiments. Biotechniques. 1992 Feb;12(2):231–232. [PubMed] [Google Scholar]
  30. Maquat L. E., Kinniburgh A. J. A beta zero-thalassemic beta-globin RNA that is labile in bone marrow cells is relatively stable in HeLa cells. Nucleic Acids Res. 1985 Apr 25;13(8):2855–2867. doi: 10.1093/nar/13.8.2855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Myers J. C., Chu M. L., Faro S. H., Clark W. J., Prockop D. J., Ramirez F. Cloning a cDNA for the pro-alpha 2 chain of human type I collagen. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3516–3520. doi: 10.1073/pnas.78.6.3516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Myers R. M., Larin Z., Maniatis T. Detection of single base substitutions by ribonuclease cleavage at mismatches in RNA:DNA duplexes. Science. 1985 Dec 13;230(4731):1242–1246. doi: 10.1126/science.4071043. [DOI] [PubMed] [Google Scholar]
  33. Nakahashi Y., Fujita H., Taketani S., Ishida N., Kappas A., Sassa S. The molecular defect of ferrochelatase in a patient with erythropoietic protoporphyria. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):281–285. doi: 10.1073/pnas.89.1.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Niwa M., Berget S. M. Mutation of the AAUAAA polyadenylation signal depresses in vitro splicing of proximal but not distal introns. Genes Dev. 1991 Nov;5(11):2086–2095. doi: 10.1101/gad.5.11.2086. [DOI] [PubMed] [Google Scholar]
  35. Pavlin D., Lichtler A. C., Bedalov A., Kream B. E., Harrison J. R., Thomas H. F., Gronowicz G. A., Clark S. H., Woody C. O., Rowe D. W. Differential utilization of regulatory domains within the alpha 1(I) collagen promoter in osseous and fibroblastic cells. J Cell Biol. 1992 Jan;116(1):227–236. doi: 10.1083/jcb.116.1.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Peterkofsky B., Diegelmann R. Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. Biochemistry. 1971 Mar 16;10(6):988–994. doi: 10.1021/bi00782a009. [DOI] [PubMed] [Google Scholar]
  37. Pihlajaniemi T., Dickson L. A., Pope F. M., Korhonen V. R., Nicholls A., Prockop D. J., Myers J. C. Osteogenesis imperfecta: cloning of a pro-alpha 2(I) collagen gene with a frameshift mutation. J Biol Chem. 1984 Nov 10;259(21):12941–12944. [PubMed] [Google Scholar]
  38. Robberson B. L., Cote G. J., Berget S. M. Exon definition may facilitate splice site selection in RNAs with multiple exons. Mol Cell Biol. 1990 Jan;10(1):84–94. doi: 10.1128/mcb.10.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shapiro J. R., Stover M. L., Burn V. E., McKinstry M. B., Burshell A. L., Chipman S. D., Rowe D. W. An osteopenic nonfracture syndrome with features of mild osteogenesis imperfecta associated with the substitution of a cysteine for glycine at triple helix position 43 in the pro alpha 1(I) chain of type I collagen. J Clin Invest. 1992 Feb;89(2):567–573. doi: 10.1172/JCI115622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Spotila L. D., Constantinou C. D., Sereda L., Ganguly A., Riggs B. L., Prockop D. J. 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 U S A. 1991 Jun 15;88(12):5423–5427. doi: 10.1073/pnas.88.12.5423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Steinmann B., Rao V. H., Vogel A., Bruckner P., Gitzelmann R., Byers P. H. Cysteine in the triple-helical domain of one allelic product of the alpha 1(I) gene of type I collagen produces a lethal form of osteogenesis imperfecta. J Biol Chem. 1984 Sep 10;259(17):11129–11138. [PubMed] [Google Scholar]
  42. Stolow D. T., Berget S. M. Identification of nuclear proteins that specifically bind to RNAs containing 5' splice sites. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):320–324. doi: 10.1073/pnas.88.2.320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sykes B., Puddle B., Francis M., Smith R. The estimation of two collagens from human dermis by interrupted gel electrophoresis. Biochem Biophys Res Commun. 1976 Oct 18;72(4):1472–1480. doi: 10.1016/s0006-291x(76)80180-5. [DOI] [PubMed] [Google Scholar]
  44. Talerico M., Berget S. M. Effect of 5' splice site mutations on splicing of the preceding intron. Mol Cell Biol. 1990 Dec;10(12):6299–6305. doi: 10.1128/mcb.10.12.6299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tromp G., Prockop D. J. 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. doi: 10.1073/pnas.85.14.5254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Urlaub G., Mitchell P. J., Ciudad C. J., Chasin L. A. Nonsense mutations in the dihydrofolate reductase gene affect RNA processing. Mol Cell Biol. 1989 Jul;9(7):2868–2880. doi: 10.1128/mcb.9.7.2868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Weil D., D'Alessio M., Ramirez F., Eyre D. R. Structural and functional characterization of a splicing mutation in the pro-alpha 2(I) collagen gene of an Ehlers-Danlos type VII patient. J Biol Chem. 1990 Sep 15;265(26):16007–16011. [PubMed] [Google Scholar]
  48. Weil D., D'Alessio M., Ramirez F., Steinmann B., Wirtz M. K., Glanville R. W., Hollister D. W. Temperature-dependent expression of a collagen splicing defect in the fibroblasts of a patient with Ehlers-Danlos syndrome type VII. J Biol Chem. 1989 Oct 5;264(28):16804–16809. [PubMed] [Google Scholar]
  49. Weil D., D'Alessio M., Ramirez F., de Wet W., Cole W. G., Chan D., Bateman J. F. A base substitution in the exon of a collagen gene causes alternative splicing and generates a structurally abnormal polypeptide in a patient with Ehlers-Danlos syndrome type VII. EMBO J. 1989 Jun;8(6):1705–1710. doi: 10.1002/j.1460-2075.1989.tb03562.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Wenstrup R. J., Shrago-Howe A. W., Lever L. W., Phillips C. L., Byers P. H., Cohn D. H. The effects of different cysteine for glycine substitutions within alpha 2(I) chains. Evidence of distinct structural domains within the type I collagen triple helix. J Biol Chem. 1991 Feb 5;266(4):2590–2594. [PubMed] [Google Scholar]
  51. Willing M. C., Cohn D. H., Byers P. H. Frameshift mutation near the 3' end of the COL1A1 gene of type I collagen predicts an elongated Pro alpha 1(I) chain and results in osteogenesis imperfecta type I. J Clin Invest. 1990 Jan;85(1):282–290. doi: 10.1172/JCI114424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Willing M. C., Pruchno C. J., Atkinson M., Byers P. H. 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] [Google Scholar]

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