Mouse DNA polymerase alpha. Subunit structure and identification of a species with associated exonuclease.

Y. C. Chen, E. W. Bohn, Stephen Planck, S. H. Wilson

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Abstract

Two species of alpha-polymerase with very similar catalytic properties have been purified to near homogeneity from a soluble protein fraction of mouse myeloma. Sedimentation analysis in 0.5 M salt-containing glycerol gradients indicated that both species had a native Mr of about 190,000. Each species contained nonidentical subunits with apparent molecular weights of about 47,000 and 54,000. Subunits of Mr = approximately 50,000 had been found previously in calf thymus alpha-polymerase (Holmes, A. M., Hesslewood, I. P., and Johnston, I. R. (1974) Eur. J. Biochem. 43, 487-499; (1976) Eur. J. Biochem. 62, 229-235). Tryptic peptide mapping failed to reveal primary structure homology between the subunits of the two enzymes. Thus, the two alpha-polymerases are clearly different species. These two enzymes are further distinguished by the fact that one of them has associated exonuclease activities. One activity degraded single-stranded DNA to mononucleotides in the 3' leads to 5' direction and acted distributively. The other exonuclease activity also degraded single-stranded DNA to mononucleotides, but this degradation was in the 5' leads to 3' direction in a processive fashion. Both exonuclease activities co-migrated with the polymerase activity during the final purification step of polyacrylamide gradient gel electrophoresis, which yielded the essentially homogenous alpha-polymerase, and also during sedimentation of the purified enzyme through a high salt glycerol gradient.

Original languageEnglish (US)
Pages (from-to)11678-11687
Number of pages10
JournalJournal of Biological Chemistry
Volume254
Issue number22
StatePublished - Nov 25 1979
Externally publishedYes

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DNA Polymerase I
Exonucleases
Single-Stranded DNA
Sedimentation
Glycerol
Enzymes
Salts
Thymus
Peptide Mapping
Electrophoresis
Thymus Gland
Purification
Polyacrylamide Gel Electrophoresis
Molecular Weight
Gels
Molecular weight
Degradation
Peptides
Proteins
Direction compound

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mouse DNA polymerase alpha. Subunit structure and identification of a species with associated exonuclease. / Chen, Y. C.; Bohn, E. W.; Planck, Stephen; Wilson, S. H.

In: Journal of Biological Chemistry, Vol. 254, No. 22, 25.11.1979, p. 11678-11687.

Research output: Contribution to journalArticle

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