Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1982 Feb 11;10(3):935–946. doi: 10.1093/nar/10.3.935

Structural homology among calf thymus alpha-polymerase polypeptides.

W Albert, F Grummt, U Hübscher, S H Wilson
PMCID: PMC326212  PMID: 7063421

Abstract

A sample of highly purified calf thymus alpha-polymerase contained an abundant 118,000 Mr polypeptide as well as five lower molecular weight polypeptides in the range of 54,000- to 64,000-Mr. This 118,000-Mr polypeptide was capable of DNA polymerase activity, as revealed by in situ assay after SDS-polyacrylamide gel electrophoresis. Tryptic peptide mapping indicated that the 118,000-Mr polypeptide shared extensive primary structure homology with 57,000-, 58,000- and 64,000-Mr polypeptides and some limited homology with 54,000- and 56,000-Mr polypeptides. This is the first evidence that lower and higher Mr polypeptides of purified calf thymus alpha-polymerase share sequence homology; these results are interpreted in the context of a model that predicts the existence of a common precursor with molecular weight greater than 140,000.

Full text

PDF
935

Images in this article

937Image
on p.937
938Image
on p.938
940Image
on p.940
942Image
on p.942
943Image
on p.943

Selected References

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

  1. Banks G. R., Boezi J. A., Lehman I. R. A high molecular weight DNA polymerase from Drosophila melanogaster embryos. Purification, structure, and partial characterization. J Biol Chem. 1979 Oct 10;254(19):9886–9892. [PubMed] [Google Scholar]
  2. Banks G. R., Holloman W. K., Kairis M. V., Spanos A., Yarranton G. T. A DNA polymerase from Ustilago maydis. 1. Purification and properties of the polymerase activity. Eur J Biochem. 1976 Feb 2;62(1):131–142. doi: 10.1111/j.1432-1033.1976.tb10106.x. [DOI] [PubMed] [Google Scholar]
  3. Brakel C. L., Blumenthal A. B. Multiple forms of Drosophila embryo DNA polymerase: evidence for proteolytic conversion. Biochemistry. 1977 Jul 12;16(14):3137–3143. doi: 10.1021/bi00633a016. [DOI] [PubMed] [Google Scholar]
  4. Chen Y. C., Bohn E. W., Planck S. R., Wilson S. H. Mouse DNA polymerase alpha. Subunit structure and identification of a species with associated exonuclease. J Biol Chem. 1979 Nov 25;254(22):11678–11687. [PubMed] [Google Scholar]
  5. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  6. Elder J. H., Pickett R. A., 2nd, Hampton J., Lerner R. A. Radioiodination of proteins in single polyacrylamide gel slices. Tryptic peptide analysis of all the major members of complex multicomponent systems using microgram quantities of total protein. J Biol Chem. 1977 Sep 25;252(18):6510–6515. [PubMed] [Google Scholar]
  7. Fisher P. A., Korn D. DNA polymerase-alpha. Purification and structural characterization of the near homogeneous enzyme from human KB cells. J Biol Chem. 1977 Sep 25;252(18):6528–6535. [PubMed] [Google Scholar]
  8. Grummt F., Waltl G., Jantzen H. M., Hamprecht K., Huebscher U., Kuenzle C. C. Diadenosine 5',5'''-P1,P4-tetraphosphate, a ligand of the 57-kilodalton subunit of DNA polymerase alpha. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6081–6085. doi: 10.1073/pnas.76.12.6081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hesslewood I. P., Holmes A. M., Wakeling W. F., Johnston I. R. Studies on the purification and properties of a 6.8-S DNA polymerase activity found in calf-thymus DNA polymerase-alpha fraction. Eur J Biochem. 1978 Mar;84(1):123–131. doi: 10.1111/j.1432-1033.1978.tb12148.x. [DOI] [PubMed] [Google Scholar]
  10. Holmes A. M., Hesslewood I. P., Johnston I. R. Evidence that DNA polymerase-alpha of calf thymus contains a subunit of molecular weight 155000. Eur J Biochem. 1976 Feb 16;62(2):229–235. doi: 10.1111/j.1432-1033.1976.tb10152.x. [DOI] [PubMed] [Google Scholar]
  11. Holmes A. M., Johnston I. R. DNA polymerases of eukaryotes. FEBS Lett. 1975 Dec 15;60(2):233–243. doi: 10.1016/0014-5793(75)80721-6. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. McKune K., Holmes A. M. Further studies on partially purified calf thymus DNA polymerase a. Nucleic Acids Res. 1979 Jul 25;6(10):3341–3352. doi: 10.1093/nar/6.10.3341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mechali M., Abadiedebat J., de Recondo A. M. Eukaryotic DNA polymerase alpha. Structural analysis of the enzyme from regenerating rat liver. J Biol Chem. 1980 Mar 10;255(5):2114–2122. [PubMed] [Google Scholar]
  15. Planck S. R., Tanabe K., Wilson S. H. Distinction between mouse DNA polymerases alpha and beta by tryptic peptide mapping. Nucleic Acids Res. 1980 Jun 25;8(12):2771–2782. doi: 10.1093/nar/8.12.2771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Planck S. R., Wilson S. H. Studies on the structure of mouse helix-destabilizing protein-1. DNA binding and controlled proteolysis with trypsin. J Biol Chem. 1980 Dec 10;255(23):11547–11556. [PubMed] [Google Scholar]
  17. Spanos A., Sedgwick S. G., Yarranton G. T., Hübscher U., Banks G. R. Detection of the catalytic activities of DNA polymerases and their associated exonucleases following SDS-polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Apr 24;9(8):1825–1839. doi: 10.1093/nar/9.8.1825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Villani G., Fay P. J., Bambara R. A., Lehman I. R. Elongation of RNA-primed DNA templates by DNA polymerase alpha from Drosophila melanogaster embryos. J Biol Chem. 1981 Aug 10;256(15):8202–8207. [PubMed] [Google Scholar]
  19. Villani G., Sauer B., Lehman I. R. DNA polymerase alpha from Drosophila melanogaster embryos. Subunit structure. J Biol Chem. 1980 Oct 10;255(19):9479–9483. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES