Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1995 Jan 1;128(1):223–237. doi: 10.1083/jcb.128.1.223

Human COL2A1-directed SV40 T antigen expression in transgenic and chimeric mice results in abnormal skeletal development

PMCID: PMC2120328  PMID: 7822417

Abstract

The ability of SV40 T antigen to cause abnormalities in cartilage development in transgenic mice and chimeras has been tested. The cis- regulatory elements of the COL2A1 gene were used to target expression of SV40 T antigen to differentiating chondrocytes in transgenic mice and chimeras derived from embryonal stem (ES) cells bearing the same transgene. The major phenotypic consequences of transgenic (pAL21) expression are malformed skeleton, disproportionate dwarfism, and perinatal/neonatal death. Expression of T antigen was tissue specific and in the main characteristic of the mouse alpha 1(II) collagen gene. Chondrocyte densities and levels of alpha 1(II) collagen mRNAs were reduced in the transgenic mice. Islands of cells which express cartilage characteristic genes such as type IIB procollagen, long form alpha 1(IX) collagen, alpha 2(XI) collagen, and aggrecan were found in the articular and growth cartilages of pAL21 chimeric fetuses and neonates. But these cells, which were expressing T antigen, were not properly organized into columns of proliferating chondrocytes. Levels of alpha 1(II) collagen mRNA were reduced in these chondrocytes. In addition, these cells did not express type X collagen, a marker for hypertrophic chondrocytes. The skeletal abnormality in pAL21 mice may therefore be due to a retardation of chondrocyte maturation or an impaired ability of chondrocytes to complete terminal differentiation and an associated paucity of some cartilage matrix components.

Full Text

The Full Text of this article is available as a PDF (9.0 MB).

Selected References

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

  1. Adams J. M., Cory S. Transgenic models of tumor development. Science. 1991 Nov 22;254(5035):1161–1167. doi: 10.1126/science.1957168. [DOI] [PubMed] [Google Scholar]
  2. Adams S. L., Boettiger D., Focht R. J., Holtzer H., Pacifici M. Regulation of the synthesis of extracellular matrix components in chondroblasts transformed by a temperature-sensitive mutant of Rous sarcoma virus. Cell. 1982 Sep;30(2):373–384. doi: 10.1016/0092-8674(82)90235-5. [DOI] [PubMed] [Google Scholar]
  3. Ahmad N. N., Ala-Kokko L., Knowlton R. G., Jimenez S. A., Weaver E. J., Maguire J. I., Tasman W., Prockop D. J. Stop codon in the procollagen II gene (COL2A1) in a family with the Stickler syndrome (arthro-ophthalmopathy). Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6624–6627. doi: 10.1073/pnas.88.15.6624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ahmad N. N., McDonald-McGinn D. M., Zackai E. H., Knowlton R. G., LaRossa D., DiMascio J., Prockop D. J. A second mutation in the type II procollagen gene (COL2AI) causing stickler syndrome (arthro-ophthalmopathy) is also a premature termination codon. Am J Hum Genet. 1993 Jan;52(1):39–45. [PMC free article] [PubMed] [Google Scholar]
  5. Ala-Kokko L., Baldwin C. T., Moskowitz R. W., Prockop D. J. Single base mutation in the type II procollagen gene (COL2A1) as a cause of primary osteoarthritis associated with a mild chondrodysplasia. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6565–6568. doi: 10.1073/pnas.87.17.6565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bonaventure J., Stanescu R., Stanescu V., Allain J. C., Muriel M. P., Ginisty D., Maroteaux P. Type II collagen defect in two sibs with the Goldblatt syndrome, a chondrodysplasia with dentinogenesis imperfecta, and joint laxity. Am J Med Genet. 1992 Dec 1;44(6):738–753. doi: 10.1002/ajmg.1320440607. [DOI] [PubMed] [Google Scholar]
  7. Bornstein P., Sage H. Regulation of collagen gene expression. Prog Nucleic Acid Res Mol Biol. 1989;37:67–106. doi: 10.1016/s0079-6603(08)60695-9. [DOI] [PubMed] [Google Scholar]
  8. Brenner R. E., Nerlich A., Kirchner F., Mörike M., Terinde R., Teller W. M. Proliferation and collagen biosynthesis of osteoblasts and chondrocytes in short rib syndrome type beemer. Am J Med Genet. 1993 Jun 15;46(5):584–591. doi: 10.1002/ajmg.1320460525. [DOI] [PubMed] [Google Scholar]
  9. Brown D. M., Nichols B. E., Weingeist T. A., Sheffield V. C., Kimura A. E., Stone E. M. Procollagen II gene mutation in Stickler syndrome. Arch Ophthalmol. 1992 Nov;110(11):1589–1593. doi: 10.1001/archopht.1992.01080230089027. [DOI] [PubMed] [Google Scholar]
  10. Butel J. S., Jarvis D. L. The plasma-membrane-associated form of SV40 large tumor antigen: biochemical and biological properties. Biochim Biophys Acta. 1986 Oct 28;865(2):171–195. doi: 10.1016/0304-419x(86)90027-2. [DOI] [PubMed] [Google Scholar]
  11. Cheah K. S., Lau E. T., Au P. K., Tam P. P. Expression of the mouse alpha 1(II) collagen gene is not restricted to cartilage during development. Development. 1991 Apr;111(4):945–953. doi: 10.1242/dev.111.4.945. [DOI] [PubMed] [Google Scholar]
  12. Cheah K. S., Stoker N. G., Griffin J. R., Grosveld F. G., Solomon E. Identification and characterization of the human type II collagen gene (COL2A1). Proc Natl Acad Sci U S A. 1985 May;82(9):2555–2559. doi: 10.1073/pnas.82.9.2555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cole W. G., Hall R. K., Rogers J. G. The clinical features of spondyloepiphyseal dysplasia congenita resulting from the substitution of glycine 997 by serine in the alpha 1(II) chain of type II collagen. J Med Genet. 1993 Jan;30(1):27–35. doi: 10.1136/jmg.30.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. DeLucia A. L., Lewton B. A., Tjian R., Tegtmeyer P. Topography of simian virus 40 A protein-DNA complexes: arrangement of pentanucleotide interaction sites at the origin of replication. J Virol. 1983 Apr;46(1):143–150. doi: 10.1128/jvi.46.1.143-150.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fanning E., Knippers R. Structure and function of simian virus 40 large tumor antigen. Annu Rev Biochem. 1992;61:55–85. doi: 10.1146/annurev.bi.61.070192.000415. [DOI] [PubMed] [Google Scholar]
  16. Francomano C. A., Liberfarb R. M., Hirose T., Maumenee I. H., Streeten E. A., Meyers D. A., Pyeritz R. E. The Stickler syndrome: evidence for close linkage to the structural gene for type II collagen. Genomics. 1987 Dec;1(4):293–296. doi: 10.1016/0888-7543(87)90027-9. [DOI] [PubMed] [Google Scholar]
  17. Gallo G. J., Gruda M. C., Manuppello J. R., Alwine J. C. Activity of simian DNA-binding factors is altered in the presence of simian virus 40 (SV40) early proteins: characterization of factors binding to elements involved in activation of the SV40 late promoter. J Virol. 1990 Jan;64(1):173–184. doi: 10.1128/jvi.64.1.173-184.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Garofalo S., Metsäranta M., Ellard J., Smith C., Horton W., Vuorio E., de Crombrugghe B. Assembly of cartilage collagen fibrils is disrupted by overexpression of normal type II collagen in transgenic mice. Proc Natl Acad Sci U S A. 1993 May 1;90(9):3825–3829. doi: 10.1073/pnas.90.9.3825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Garofalo S., Vuorio E., Metsaranta M., Rosati R., Toman D., Vaughan J., Lozano G., Mayne R., Ellard J., Horton W. Reduced amounts of cartilage collagen fibrils and growth plate anomalies in transgenic mice harboring a glycine-to-cysteine mutation in the mouse type II procollagen alpha 1-chain gene. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9648–9652. doi: 10.1073/pnas.88.21.9648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hanahan D. Dissecting multistep tumorigenesis in transgenic mice. Annu Rev Genet. 1988;22:479–519. doi: 10.1146/annurev.ge.22.120188.002403. [DOI] [PubMed] [Google Scholar]
  21. Hanahan D. Heritable formation of pancreatic beta-cell tumours in transgenic mice expressing recombinant insulin/simian virus 40 oncogenes. Nature. 1985 May 9;315(6015):115–122. doi: 10.1038/315115a0. [DOI] [PubMed] [Google Scholar]
  22. Horton W. A., Machado M. A., Ellard J., Campbell D., Bartley J., Ramirez F., Vitale E., Lee B. Characterization of a type II collagen gene (COL2A1) mutation identified in cultured chondrocytes from human hypochondrogenesis. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4583–4587. doi: 10.1073/pnas.89.10.4583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Horton W. E., Jr, Cleveland J., Rapp U., Nemuth G., Bolander M., Doege K., Yamada Y., Hassell J. R. An established rat cell line expressing chondrocyte properties. Exp Cell Res. 1988 Oct;178(2):457–468. doi: 10.1016/0014-4827(88)90414-4. [DOI] [PubMed] [Google Scholar]
  24. Horton W., Miyashita T., Kohno K., Hassell J. R., Yamada Y. Identification of a phenotype-specific enhancer in the first intron of the rat collagen II gene. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8864–8868. doi: 10.1073/pnas.84.24.8864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Imai S., Takano T. Loss of collagenase gene expression in immortalized clones of SV40 T antigen-transformed human diploid fibroblasts. Biochem Biophys Res Commun. 1992 Nov 30;189(1):148–153. doi: 10.1016/0006-291x(92)91537-z. [DOI] [PubMed] [Google Scholar]
  26. Jensen N. A., Smith G. M., Shine H. D., Garvey J. S., Hood L. Distinct hypomyelinated phenotypes in MBP-SV40 large T transgenic mice. J Neurosci Res. 1993 Feb 15;34(3):257–264. doi: 10.1002/jnr.490340302. [DOI] [PubMed] [Google Scholar]
  27. Karaplis A. C., Luz A., Glowacki J., Bronson R. T., Tybulewicz V. L., Kronenberg H. M., Mulligan R. C. Lethal skeletal dysplasia from targeted disruption of the parathyroid hormone-related peptide gene. Genes Dev. 1994 Feb 1;8(3):277–289. doi: 10.1101/gad.8.3.277. [DOI] [PubMed] [Google Scholar]
  28. Kessel M., Gruss P. Homeotic transformations of murine vertebrae and concomitant alteration of Hox codes induced by retinoic acid. Cell. 1991 Oct 4;67(1):89–104. doi: 10.1016/0092-8674(91)90574-i. [DOI] [PubMed] [Google Scholar]
  29. Knowlton R. G., Katzenstein P. L., Moskowitz R. W., Weaver E. J., Malemud C. J., Pathria M. N., Jimenez S. A., Prockop D. J. Genetic linkage of a polymorphism in the type II procollagen gene (COL2A1) to primary osteoarthritis associated with mild chondrodysplasia. N Engl J Med. 1990 Feb 22;322(8):526–530. doi: 10.1056/NEJM199002223220807. [DOI] [PubMed] [Google Scholar]
  30. Kong R. Y., Kwan K. M., Lau E. T., Thomas J. T., Boot-Handford R. P., Grant M. E., Cheah K. S. Intron-exon structure, alternative use of promoter and expression of the mouse collagen X gene, Col10a-1. Eur J Biochem. 1993 Apr 1;213(1):99–111. doi: 10.1111/j.1432-1033.1993.tb17739.x. [DOI] [PubMed] [Google Scholar]
  31. Lee B., Vissing H., Ramirez F., Rogers D., Rimoin D. Identification of the molecular defect in a family with spondyloepiphyseal dysplasia. Science. 1989 May 26;244(4907):978–980. doi: 10.1126/science.2543071. [DOI] [PubMed] [Google Scholar]
  32. Liu C. Y., Olsen B. R., Kao W. W. Developmental patterns of two alpha 1(IX) collagen mRNA isoforms in mouse. Dev Dyn. 1993 Oct;198(2):150–157. doi: 10.1002/aja.1001980208. [DOI] [PubMed] [Google Scholar]
  33. Lovell-Badge R. H., Bygrave A., Bradley A., Robertson E., Tilly R., Cheah K. S. Tissue-specific expression of the human type II collagen gene in mice. Proc Natl Acad Sci U S A. 1987 May;84(9):2803–2807. doi: 10.1073/pnas.84.9.2803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mallein-Gerin F., Olsen B. R. Expression of simian virus 40 large T (tumor) oncogene in mouse chondrocytes induces cell proliferation without loss of the differentiated phenotype. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3289–3293. doi: 10.1073/pnas.90.8.3289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Metsäranta M., Garofalo S., Decker G., Rintala M., de Crombrugghe B., Vuorio E. Chondrodysplasia in transgenic mice harboring a 15-amino acid deletion in the triple helical domain of pro alpha 1(II) collagen chain. J Cell Biol. 1992 Jul;118(1):203–212. doi: 10.1083/jcb.118.1.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Muragaki Y., Nishimura I., Henney A., Ninomiya Y., Olsen B. R. The alpha 1 (IX) collagen gene gives rise to two different transcripts in both mouse embryonic and human fetal RNA. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2400–2404. doi: 10.1073/pnas.87.7.2400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ng L. J., Tam P. P., Cheah K. S. Preferential expression of alternatively spliced mRNAs encoding type II procollagen with a cysteine-rich amino-propeptide in differentiating cartilage and nonchondrogenic tissues during early mouse development. Dev Biol. 1993 Oct;159(2):403–417. doi: 10.1006/dbio.1993.1251. [DOI] [PubMed] [Google Scholar]
  38. Palotie A., Väisänen P., Ott J., Ryhänen L., Elima K., Vikkula M., Cheah K., Vuorio E., Peltonen L. Predisposition to familial osteoarthrosis linked to type II collagen gene. Lancet. 1989 Apr 29;1(8644):924–927. doi: 10.1016/s0140-6736(89)92507-5. [DOI] [PubMed] [Google Scholar]
  39. Parker M. I., Smith A. A., Mundell K., Collins M., Boast S., Ramirez F. The abolition of collagen gene expression in SV40-transformed fibroblasts is associated with trans-acting factor switching. Nucleic Acids Res. 1992 Nov 11;20(21):5825–5830. doi: 10.1093/nar/20.21.5825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Poole A. R., Pidoux I., Reiner A., Rosenberg L., Hollister D., Murray L., Rimoin D. Kniest dysplasia is characterized by an apparent abnormal processing of the C-propeptide of type II cartilage collagen resulting in imperfect fibril assembly. J Clin Invest. 1988 Feb;81(2):579–589. doi: 10.1172/JCI113356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Ryan M. C., Sandell L. J. Differential expression of a cysteine-rich domain in the amino-terminal propeptide of type II (cartilage) procollagen by alternative splicing of mRNA. J Biol Chem. 1990 Jun 25;265(18):10334–10339. [PubMed] [Google Scholar]
  42. Ryan M. C., Sieraski M., Sandell L. J. The human type II procollagen gene: identification of an additional protein-coding domain and location of potential regulatory sequences in the promoter and first intron. Genomics. 1990 Sep;8(1):41–48. doi: 10.1016/0888-7543(90)90224-i. [DOI] [PubMed] [Google Scholar]
  43. Sandell L. J., Morris N., Robbins J. R., Goldring M. B. Alternatively spliced type II procollagen mRNAs define distinct populations of cells during vertebral development: differential expression of the amino-propeptide. J Cell Biol. 1991 Sep;114(6):1307–1319. doi: 10.1083/jcb.114.6.1307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Sladek T. L., Jacobberger J. W. Simian virus 40 large T-antigen expression decreases the G1 and increases the G2 + M cell cycle phase durations in exponentially growing cells. J Virol. 1992 Feb;66(2):1059–1065. doi: 10.1128/jvi.66.2.1059-1065.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Spranger J., Maroteaux P. The lethal osteochondrodysplasias. Adv Hum Genet. 1990;19:1-103, 331-2. doi: 10.1007/978-1-4757-9065-8_1. [DOI] [PubMed] [Google Scholar]
  46. Stahl H., Knippers R. The simian virus 40 large tumor antigen. Biochim Biophys Acta. 1987 Oct 9;910(1):1–10. doi: 10.1016/0167-4781(87)90088-1. [DOI] [PubMed] [Google Scholar]
  47. Swiderski R. E., Solursh M. Localization of type II collagen, long form alpha 1(IX) collagen, and short form alpha 1(IX) collagen transcripts in the developing chick notochord and axial skeleton. Dev Dyn. 1992 Jun;194(2):118–127. doi: 10.1002/aja.1001940205. [DOI] [PubMed] [Google Scholar]
  48. Thenet S., Benya P. D., Demignot S., Feunteun J., Adolphe M. SV40-immortalization of rabbit articular chondrocytes: alteration of differentiated functions. J Cell Physiol. 1992 Jan;150(1):158–167. doi: 10.1002/jcp.1041500121. [DOI] [PubMed] [Google Scholar]
  49. Tiller G. E., Rimoin D. L., Murray L. W., Cohn D. H. Tandem duplication within a type II collagen gene (COL2A1) exon in an individual with spondyloepiphyseal dysplasia. Proc Natl Acad Sci U S A. 1990 May;87(10):3889–3893. doi: 10.1073/pnas.87.10.3889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Vandenberg P., Khillan J. S., Prockop D. J., Helminen H., Kontusaari S., Ala-Kokko L. Expression of a partially deleted gene of human type II procollagen (COL2A1) in transgenic mice produces a chondrodysplasia. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7640–7644. doi: 10.1073/pnas.88.17.7640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Vikkula M., Metsäranta M., Syvänen A. C., Ala-Kokko L., Vuorio E., Peltonen L. Structural analysis of the regulatory elements of the type-II procollagen gene. Conservation of promoter and first intron sequences between human and mouse. Biochem J. 1992 Jul 1;285(Pt 1):287–294. doi: 10.1042/bj2850287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Vikkula M., Palotie A., Ritvaniemi P., Ott J., Ala-Kokko L., Sievers U., Aho K., Peltonen L. Early-onset osteoarthritis linked to the type II procollagen gene. Detailed clinical phenotype and further analyses of the gene. Arthritis Rheum. 1993 Mar;36(3):401–409. doi: 10.1002/art.1780360317. [DOI] [PubMed] [Google Scholar]
  53. Vikkula M., Ritvaniemi P., Vuorio A. F., Kaitila I., Ala-Kokko L., Peltonen L. A mutation in the amino-terminal end of the triple helix of type II collagen causing severe osteochondrodysplasia. Genomics. 1993 Apr;16(1):282–285. doi: 10.1006/geno.1993.1179. [DOI] [PubMed] [Google Scholar]
  54. Vissing H., D'Alessio M., Lee B., Ramirez F., Godfrey M., Hollister D. W. Glycine to serine substitution in the triple helical domain of pro-alpha 1 (II) collagen results in a lethal perinatal form of short-limbed dwarfism. J Biol Chem. 1989 Nov 5;264(31):18265–18267. [PubMed] [Google Scholar]
  55. Winterpacht A., Hilbert M., Schwarze U., Mundlos S., Spranger J., Zabel B. U. Kniest and Stickler dysplasia phenotypes caused by collagen type II gene (COL2A1) defect. Nat Genet. 1993 Apr;3(4):323–326. doi: 10.1038/ng0493-323. [DOI] [PubMed] [Google Scholar]
  56. Wood A., Ashhurst D. E., Corbett A., Thorogood P. The transient expression of type II collagen at tissue interfaces during mammalian craniofacial development. Development. 1991 Apr;111(4):955–968. doi: 10.1242/dev.111.4.955. [DOI] [PubMed] [Google Scholar]
  57. al-Ubaidi M. R., Hollyfield J. G., Overbeek P. A., Baehr W. Photoreceptor degeneration induced by the expression of simian virus 40 large tumor antigen in the retina of transgenic mice. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1194–1198. doi: 10.1073/pnas.89.4.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES