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. 1983;2(8):1223–1227. doi: 10.1002/j.1460-2075.1983.tb01573.x

Procollagen alpha 2(I) mRNA in dermatosparactic fibroblasts: evidence for post-transcriptional regulation.

G Lozano 1, O Helle 1, P K Müller 1
PMCID: PMC555264  PMID: 10872312

Abstract

Dermatosparaxis is a genetic defect of connective tissues found in man and animals. Dermatosparactic sheep fibroblasts fail to cleave off the aminopropeptide of procollagen, this makes the feedback regulation less effective and leads to an elevated production of collagen. We have isolated the total RNA from dermatosparactic and normal sheep fibroblasts and then compared the amount of mRNA sequences complementary to alpha 2(I) genomic DNA. Using methyl mercury gels, the Northern hybridization procedure and dotting hybridization, we were able to show that the levels of mRNA specific for alpha 2(I) collagen were only marginally different in both fibroblast strains. Furthermore, the amount of collagen mRNA translatable in a cell-free translation system was the same in affected and non-affected sheep cells. Our data provide further evidence for a post-transcriptional control of collagen synthesis in fibroblasts.

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Selected References

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

  1. Adams S. L., Sobel M. E., Howard B. H., Olden K., Yamada K. M., de Crombrugghe B., Pastan I. Levels of translatable mRNAs for cell surface protein, collagen precursors, and two membrane proteins are altered in Rous sarcoma virus-transformed chick embryo fibroblasts. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3399–3403. doi: 10.1073/pnas.74.8.3399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bellamy G., Bornstein P. Evidence for procollagen, a biosynthetic precursors of collagen. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1138–1142. doi: 10.1073/pnas.68.6.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. Boyd C. D., Tolstoshev P., Schafer M. P., Trapnell B. C., Coon H. C., Kretschmer P. J., Nienhuis A. W., Crystal R. G. Isolation and characterization of a 15-kilobase genomic sequence coding for part of the Pro alpha 2 chain of sheep type I collagen. J Biol Chem. 1980 Apr 10;255(7):3212–3220. [PubMed] [Google Scholar]
  6. Fessler J. H., Fessler L. I. Biosynthesis of procollagen. Annu Rev Biochem. 1978;47:129–162. doi: 10.1146/annurev.bi.47.070178.001021. [DOI] [PubMed] [Google Scholar]
  7. Helle O., Nes N. N. A hereditary skin defect in sheep. Acta Vet Scand. 1972;13(3):443–445. doi: 10.1186/BF03547060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Lapière C. M., Lenaers A., Kohn L. D. Procollagen peptidase: an enzyme excising the coordination peptides of procollagen. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3054–3058. doi: 10.1073/pnas.68.12.3054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. OGUR M., ROSEN G. The nucleic acids of plant tissues; the extraction and estimation of desoxypentose nucleic acid and pentose nucleic acid. Arch Biochem. 1950 Feb;25(2):262–276. [PubMed] [Google Scholar]
  13. Overbeek P. A., Merlino G. T., Peters N. K., Cohn V. H., Moore G. P., Kleinsmith L. J. Characterization of five members of the actin gene family in the sea urchin. Biochim Biophys Acta. 1981 Dec 28;656(2):195–205. doi: 10.1016/0005-2787(81)90087-3. [DOI] [PubMed] [Google Scholar]
  14. Paglia L., Wilczek J., de Leon L. D., Martin G. R., Hörlein D., Müller P. Inhibition of procollagen cell-free synthesis by amino-terminal extension peptides. Biochemistry. 1979 Oct 30;18(22):5030–5034. doi: 10.1021/bi00589a034. [DOI] [PubMed] [Google Scholar]
  15. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  16. Prockop D. J., Tuderman L. Posttranslational enzymes in the biosynthesis of collagen: extracellular enzymes. Methods Enzymol. 1982;82(Pt A):305–319. doi: 10.1016/0076-6879(82)82068-5. [DOI] [PubMed] [Google Scholar]
  17. Rowe D. W., Moen R. C., Davidson J. M., Byers P. H., Bornstein P., Palmiter R. D. Correlation of procollagen mRNA levels in normal and transformed chick embryo fibroblasts with different rates of procollagen synthesis. Biochemistry. 1978 May 2;17(9):1581–1590. doi: 10.1021/bi00602a001. [DOI] [PubMed] [Google Scholar]
  18. Sandmeyer S., Gallis B., Bornstein P. Coordinate transcriptional regulation of type I procollagen genes by Rous sarcoma virus. J Biol Chem. 1981 May 25;256(10):5022–5028. [PubMed] [Google Scholar]
  19. Sobel M. E., Yamamoto T., Adams S. L., DiLauro R., Avvedimento V. E., de Crombrugghe B., Pastan I. Construction of a recombinant bacterial plasmid containing a chick pro-alpha2 collagen gene sequence. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5846–5850. doi: 10.1073/pnas.75.12.5846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tolstoshev P., Haber R., Trapnell B. C., Crystal R. G. Procollagen messenger RNA levels and activity and collagen synthesis during the fetal development of sheep lung, tendon, and skin. J Biol Chem. 1981 Sep 25;256(18):9672–9679. [PubMed] [Google Scholar]
  21. Wiestner M., Krieg T., Hörlein D., Glanville R. W., Fietzek P., Müller P. K. Inhibiting effect of procollagen peptides on collagen biosynthesis in fibroblast cultures. J Biol Chem. 1979 Aug 10;254(15):7016–7023. [PubMed] [Google Scholar]
  22. Wiestner M., Rohde H., Helle O., Krieg T., Timpl R., Müller P. K. Low rate of procollagen conversion in dermatosparactic sheep fibroblasts is paralleled by increased synthesis of type I and type III collagens. EMBO J. 1982;1(4):513–516. doi: 10.1002/j.1460-2075.1982.tb01200.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yamamoto T., Sobel M. E., Adams S. L., Avvedimento V. E., DiLauro R., Pastan I., de Crombrugghe B., Showalter A., Pesciotta D., Fietzek P. Construction of a recombinant bacterial plasmid containing pro-alpha 1(I) collagen DNA sequences. J Biol Chem. 1980 Mar 25;255(6):2612–2615. [PubMed] [Google Scholar]

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