Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1983 Apr;80(7):1811–1815. doi: 10.1073/pnas.80.7.1811

In vitro synthesis of thymosin beta 4 encoded by rat spleen mRNA.

A W Filipowicz, B L Horecker
PMCID: PMC393699  PMID: 6572941

Abstract

Thymosin beta 4, containing 43 amino acids and acetylated at the NH2 terminus, is synthesized in vitro in a rabbit reticulocyte lysate or in a yeast protein-synthesis system in the presence of mRNA from rat spleen. The product formed was identified as beta 4 by immunoprecipitation by a specific anti-beta 4 antiserum, comigration with authentic beta 4 in NaDodSO4/polyacrylamide gel electrophoresis and in HPLC, and identity of peptide fragments. The immunoprecipitable product generated in the wheat germ protein-synthesizing system emerged slightly ahead of beta 4 in HPLC and appeared to lack the NH2-terminal acetyl group. There was no evidence for formation of a larger polypeptide precursor of beta 4 in any of the three systems used. In sucrose density gradient centrifugation, the mRNA coding for beta 4 was recovered in the 7-8S mRNA fraction.

Full text

PDF
1811

Images in this article

1811Image
on p.1811

Selected References

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

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bell G. I., Swain W. F., Pictet R., Cordell B., Goodman H. M., Rutter W. J. Nucleotide sequence of a cDNA clone encoding human preproinsulin. Nature. 1979 Nov 29;282(5738):525–527. doi: 10.1038/282525a0. [DOI] [PubMed] [Google Scholar]
  3. Bloemendal H. The vertebrate eye lens. Science. 1977 Jul 8;197(4299):127–138. doi: 10.1126/science.877544. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Comb M., Seeburg P. H., Adelman J., Eiden L., Herbert E. Primary structure of the human Met- and Leu-enkephalin precursor and its mRNA. Nature. 1982 Feb 25;295(5851):663–666. doi: 10.1038/295663a0. [DOI] [PubMed] [Google Scholar]
  6. Davies P. L., Roach A. H., Hew C. L. DNA sequence coding for an antifreeze protein precursor from winter flounder. Proc Natl Acad Sci U S A. 1982 Jan;79(2):335–339. doi: 10.1073/pnas.79.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Docherty K., Steiner D. F. Post-translational proteolysis in polypeptide hormone biosynthesis. Annu Rev Physiol. 1982;44:625–638. doi: 10.1146/annurev.ph.44.030182.003205. [DOI] [PubMed] [Google Scholar]
  8. Eisenman R., Shaikh R., Mason W. S. Identification of an avian oncovirus polyprotein in uninfected chick cells. Cell. 1978 May;14(1):89–104. doi: 10.1016/0092-8674(78)90304-5. [DOI] [PubMed] [Google Scholar]
  9. El-Dorry H. A., MacGregor J. S. Rat and rabbit liver mRNAs code for fructose 1,6-bisphosphatases that differ in molecular weight. Biochem Biophys Res Commun. 1982 Aug 31;107(4):1384–1389. doi: 10.1016/s0006-291x(82)80151-4. [DOI] [PubMed] [Google Scholar]
  10. Gedamu L., Dixon G. H. Assay of protamine messenger RNA from rainbow trout testis. J Biol Chem. 1976 Mar 10;251(5):1446–1454. [PubMed] [Google Scholar]
  11. Gedamu L., Dixon G. H. Purification and properties of biologically active rainbow trout testis protamine mRNA. J Biol Chem. 1976 Mar 10;251(5):1455–1463. [PubMed] [Google Scholar]
  12. Gubler U., Kilpatrick D. L., Seeburg P. H., Gage L. P., Udenfriend S. Detection and partial characterization of proenkephalin mRNA. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5484–5487. doi: 10.1073/pnas.78.9.5484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hannappel E., Davoust S., Horecker B. L. Isolation of peptides from calf thymus. Biochem Biophys Res Commun. 1982 Jan 15;104(1):266–271. doi: 10.1016/0006-291x(82)91969-6. [DOI] [PubMed] [Google Scholar]
  14. Hannappel E., Xu G. J., Morgan J., Hempstead J., Horecker B. L. Thymosin beta 4: a ubiquitous peptide in rat and mouse tissues. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2172–2175. doi: 10.1073/pnas.79.7.2172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kedes L. H. Histone genes and histone messengers. Annu Rev Biochem. 1979;48:837–870. doi: 10.1146/annurev.bi.48.070179.004201. [DOI] [PubMed] [Google Scholar]
  16. Kenny C., Moschera J. A., Stein S. Purification of human fibroblast interferon produced in the absence of serum by Cibacron Blue F3GA-Agarose and high-performance liquid chromatography. Methods Enzymol. 1981;78(Pt A):435–447. doi: 10.1016/0076-6879(81)78154-0. [DOI] [PubMed] [Google Scholar]
  17. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  18. Kido H., Vita A., Hannappel E., Horecker B. L. Aminoterminal acetylation of synthetic N alpha-desacetyl thymosin alpha 1. Arch Biochem Biophys. 1981 Apr 15;208(1):95–100. doi: 10.1016/0003-9861(81)90127-2. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Land H., Schütz G., Schmale H., Richter D. Nucleotide sequence of cloned cDNA encoding bovine arginine vasopressin-neurophysin II precursor. Nature. 1982 Jan 28;295(5847):299–303. doi: 10.1038/295299a0. [DOI] [PubMed] [Google Scholar]
  21. Low T. L., Hu S. K., Goldstein A. L. Complete amino acid sequence of bovine thymosin beta 4: a thymic hormone that induces terminal deoxynucleotidyl transferase activity in thymocyte populations. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1162–1166. doi: 10.1073/pnas.78.2.1162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nakanishi S., Inoue A., Kita T., Nakamura M., Chang A. C., Cohen S. N., Numa S. Nucleotide sequence of cloned cDNA for bovine corticotropin-beta-lipotropin precursor. Nature. 1979 Mar 29;278(5703):423–427. doi: 10.1038/278423a0. [DOI] [PubMed] [Google Scholar]
  23. Noda M., Furutani Y., Takahashi H., Toyosato M., Hirose T., Inayama S., Nakanishi S., Numa S. Cloning and sequence analysis of cDNA for bovine adrenal preproenkephalin. Nature. 1982 Jan 21;295(5846):202–206. doi: 10.1038/295202a0. [DOI] [PubMed] [Google Scholar]
  24. Palmiter R. D. Prevention of NH2-terminal acetylation of proteins synthesized in cell-free systems. J Biol Chem. 1977 Dec 25;252(24):8781–8783. [PubMed] [Google Scholar]
  25. 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]
  26. Rebar R. W., Miyake A., Low T. L., Goldstein A. L. Thymosin stimulates secretion of luteinizing hormone-releasing factor. Science. 1981 Nov 6;214(4521):669–671. doi: 10.1126/science.7027442. [DOI] [PubMed] [Google Scholar]
  27. Roberts J. L., Herbert E. Characterization of a common precursor to corticotropin and beta-lipotropin: cell-free synthesis of the precursor and identification of corticotropin peptides in the molecule. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4826–4830. doi: 10.1073/pnas.74.11.4826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wodnar-Filipowicz A., Szczesna E., Zan-Kowalczewska M., Muthukrishnan S., Szybiak U., Legocki A. B., Filipowicz W. 5'-Terminal 7-methylguanosine and mRNA function. The effect of enzymatic decapping and of cap analogs on translation of tobacco-mosaic-virus RNA and globin mRNA in vitro. Eur J Biochem. 1978 Dec 1;92(1):69–80. doi: 10.1111/j.1432-1033.1978.tb12724.x. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES