Abstract
Poly(A)+ RNA, isolated from a single 7-mo fetal human aorta, was used to synthesize cDNA by the RNase H method, and the cDNA was inserted into lambda gt10. Recombinant phage containing elastin sequences were identified by hybridization with cloned, exon-containing fragments of the human elastin gene. Three clones containing inserts of 3.3, 2.7, and 2.3 kilobases were selected for further analysis. Three overlapping clones containing 17.8 kilobases of the human elastin gene were also isolated from genomic libraries. Complete sequence analysis of the six clones demonstrated that: the cDNA encompassed the entire translated portion of the mRNA encoding 786 amino acids, including several unusual hydrophilic amino acid sequences not previously identified in porcine tropoelastin, exons encoding either hydrophobic or crosslinking domains in the protein alternated in the gene, and a great abundance of Alu repetitive sequences occurred throughout the introns. The data also indicated substantial alternative splicing of the mRNA. These results suggest the potential for significant variation in the precise molecular structure of the elastic fiber in the human population.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
- Burnett W., Finnigan-Bunick A., Yoon K., Rosenbloom J. Analysis of elastin gene expression in the developing chick aorta using cloned elastin cDNA. J Biol Chem. 1982 Feb 25;257(4):1569–1572. [PubMed] [Google Scholar]
- Burnett W., Rosenbloom J. Isolation and translation of elastin mRNA from chick aorta. Biochem Biophys Res Commun. 1979 Feb 14;86(3):478–484. doi: 10.1016/0006-291x(79)91739-x. [DOI] [PubMed] [Google Scholar]
- Calabretta B., Robberson D. L., Barrera-Saldaña H. A., Lambrou T. P., Saunders G. F. Genome instability in a region of human DNA enriched in Alu repeat sequences. Nature. 1982 Mar 18;296(5854):219–225. doi: 10.1038/296219a0. [DOI] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Cicila G., May M., Ornstein-Goldstein N., Indik Z., Morrow S., Yeh H. S., Rosenbloom J., Boyd C., Rosenbloom J., Yoon K. Structure of the 3' portion of the bovine elastin gene. Biochemistry. 1985 Jun 18;24(13):3075–3080. doi: 10.1021/bi00334a001. [DOI] [PubMed] [Google Scholar]
- Davidson J. M., Leslie B., Wolt T., Crystal R. G., Sandberg L. B. Characterization of a signal peptide sequence in the cell-free translation product of sheep elastin mRNA. Arch Biochem Biophys. 1982 Oct 1;218(1):31–37. doi: 10.1016/0003-9861(82)90317-4. [DOI] [PubMed] [Google Scholar]
- Davidson J. M., Shibahara S., Schafer M. P., Harrison M., Leach C., Tolstoshev P., Crystal R. G. Sheep elastin genes. Isolation and preliminary characterization of a 9.9-kilobase genomic clone. Biochem J. 1984 Jun 15;220(3):643–652. doi: 10.1042/bj2200643. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deininger P. L., Jolly D. J., Rubin C. M., Friedmann T., Schmid C. W. Base sequence studies of 300 nucleotide renatured repeated human DNA clones. J Mol Biol. 1981 Sep 5;151(1):17–33. doi: 10.1016/0022-2836(81)90219-9. [DOI] [PubMed] [Google Scholar]
- Emanuel B. S., Cannizzaro L., Ornstein-Goldstein N., Indik Z. K., Yoon K., May M., Oliver L., Boyd C., Rosenbloom J. Chromosomal localization of the human elastin gene. Am J Hum Genet. 1985 Sep;37(5):873–882. [PMC free article] [PubMed] [Google Scholar]
- Fleming W. W., Sullivan C. E., Torchia D. A. Characterization of molecular motions in 13C-labeled aortic elastin by 13C-1H magnetic double resonance. Biopolymers. 1980 Mar;19(3):597–617. doi: 10.1002/bip.1980.360190311. [DOI] [PubMed] [Google Scholar]
- Foster J. A., Bruenger E., Gray W. R., Sandberg L. B. Isolation and amino acid sequences of tropoelastin peptides. J Biol Chem. 1973 Apr 25;248(8):2876–2879. [PubMed] [Google Scholar]
- Foster J. A., Rich C. B., Fletcher S., Karr S. R., Przybyla A. Translation of chick aortic elastin messenger ribonucleic acid. Comparison to elastin synthesis in chick aorta organ culture. Biochemistry. 1980 Mar 4;19(5):857–864. doi: 10.1021/bi00546a005. [DOI] [PubMed] [Google Scholar]
- Foster J. A., Shapiro R., Voynow P., Crombie G., Faris B., Franzblau C. Isolation of soluble elastin from lathyritic chicks. Comparison to tropoelastin from copper deficient pigs. Biochemistry. 1975 Dec 2;14(24):5343–5347. doi: 10.1021/bi00695a019. [DOI] [PubMed] [Google Scholar]
- Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
- Hopp T. P., Woods K. R. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. doi: 10.1073/pnas.78.6.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Indik Z., Yoon K., Morrow S. D., Cicila G., Rosenbloom J., Rosenbloom J., Ornstein-Goldstein N. Structure of the 3' region of the human elastin gene: great abundance of Alu repetitive sequences and few coding sequences. Connect Tissue Res. 1987;16(3):197–211. doi: 10.3109/03008208709006976. [DOI] [PubMed] [Google Scholar]
- Karr S. R., Foster J. A. Primary structure of the signal peptide of tropoelastin b. J Biol Chem. 1981 Jun 25;256(12):5946–5949. [PubMed] [Google Scholar]
- Lehrman M. A., Schneider W. J., Südhof T. C., Brown M. S., Goldstein J. L., Russell D. W. Mutation in LDL receptor: Alu-Alu recombination deletes exons encoding transmembrane and cytoplasmic domains. Science. 1985 Jan 11;227(4683):140–146. doi: 10.1126/science.3155573. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mecham R. P., Morris S. L., Levy B. D., Wrenn D. S. Glucocorticoids stimulate elastin production in differentiated bovine ligament fibroblasts but do not induce elastin synthesis in undifferentiated cells. J Biol Chem. 1984 Oct 25;259(20):12414–12418. [PubMed] [Google Scholar]
- Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
- Miller E. J., Martin G. R., Mecca C. E., Piez K. A. The biosynthesis of elastin cross-links. The effect of copper deficiency and a lathyrogen. J Biol Chem. 1965 Sep;240(9):3623–3627. [PubMed] [Google Scholar]
- Orkin S. H., Michelson A. Partial deletion of the alpha-globin structural gene in human alpha-thalassaemia. Nature. 1980 Jul 31;286(5772):538–540. doi: 10.1038/286538a0. [DOI] [PubMed] [Google Scholar]
- Padgett R. A., Grabowski P. J., Konarska M. M., Seiler S., Sharp P. A. Splicing of messenger RNA precursors. Annu Rev Biochem. 1986;55:1119–1150. doi: 10.1146/annurev.bi.55.070186.005351. [DOI] [PubMed] [Google Scholar]
- Partridge S. M., Elsden D. F., Thomas J., Dorfman A., Telser A., Ho P. L. Incorporation of labelled lysine into the desmosine cross-bridges in elastin. Nature. 1966 Jan 22;209(5021):399–400. doi: 10.1038/209399b0. [DOI] [PubMed] [Google Scholar]
- Pinnell S. R., Martin G. R. The cross-linking of collagen and elastin: enzymatic conversion of lysine in peptide linkage to alpha-aminoadipic-delta-semialdehyde (allysine) by an extract from bone. Proc Natl Acad Sci U S A. 1968 Oct;61(2):708–716. doi: 10.1073/pnas.61.2.708. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ruskin B., Krainer A. R., Maniatis T., Green M. R. Excision of an intact intron as a novel lariat structure during pre-mRNA splicing in vitro. Cell. 1984 Aug;38(1):317–331. doi: 10.1016/0092-8674(84)90553-1. [DOI] [PubMed] [Google Scholar]
- Sandberg L. B., Leslie J. G., Leach C. T., Alvarez V. L., Torres A. R., Smith D. W. Elastin covalent structure as determined by solid phase amino acid sequencing. Pathol Biol (Paris) 1985 Apr;33(4):266–274. [PubMed] [Google Scholar]
- Sandberg L. B., Weissman N., Smith D. W. The purification and partial characterization of a soluble elastin-like protein from copper-deficient porcine aorta. Biochemistry. 1969 Jul;8(7):2940–2945. doi: 10.1021/bi00835a037. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmid C. W., Jelinek W. R. The Alu family of dispersed repetitive sequences. Science. 1982 Jun 4;216(4550):1065–1070. doi: 10.1126/science.6281889. [DOI] [PubMed] [Google Scholar]
- Sharp P. A. Conversion of RNA to DNA in mammals: Alu-like elements and pseudogenes. Nature. 1983 Feb 10;301(5900):471–472. doi: 10.1038/301471a0. [DOI] [PubMed] [Google Scholar]
- Yoon K., Davidson J. M., Boyd C., May M., LuValle P., Ornstein-Goldstein N., Smith J., Indik Z., Ross A., Golub E. Analysis of the 3' region of the sheep elastin gene. Arch Biochem Biophys. 1985 Sep;241(2):684–691. doi: 10.1016/0003-9861(85)90595-8. [DOI] [PubMed] [Google Scholar]
- Yoon K., May M., Goldstein N., Indik Z. K., Oliver L., Boyd C., Rosenbloom J. Characterization of a sheep elastin cDNA clone containing translated sequences. Biochem Biophys Res Commun. 1984 Jan 13;118(1):261–269. doi: 10.1016/0006-291x(84)91095-7. [DOI] [PubMed] [Google Scholar]