Studies of the domain structure of mammalian DNA polymerase β. Identification of a discrete template binding domain

A. Kumar, Steven Widen, K. R. Williams, P. Kedar, R. L. Karpel, S. H. Wilson

Research output: Contribution to journalArticle

149 Citations (Scopus)

Abstract

Characterization of the domain structure of DNA polymerase β is reported. Large scale overproduction of the rat protein in coli was achieved, and the purified recombinant protein was verified by sequencing tryptic peptides. This protein is both a single-stranded DNA binding protein and a DNA polymerase consisting of one polypeptide chain of 334 amino acids. As revealed by controlled proteolysis experiments, the protein is organized in two relatively protease-resistant segments linked by a short protease-sensitive region. One of these protease-resistant segments represents the NH2-terminal 20% of the protein. This NH2-terminal domain (of about 75 residues) has strong affinity for single-stranded nucleic acids. The other protease-resistant segment, representing the COOH-terminal domain of ~ 250 residues, does not bind to nucleic acids. Neither domain, tested as purified proteins, has substantial DNA polymerase activity. The results suggest that the NH2-terminal domain is principally responsible for the template binding activity of the intact protein.

Original languageEnglish (US)
Pages (from-to)2124-2131
Number of pages8
JournalJournal of Biological Chemistry
Volume265
Issue number4
StatePublished - Aug 29 1990
Externally publishedYes

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DNA-Directed DNA Polymerase
Peptide Hydrolases
Proteins
Nucleic Acids
Proteolysis
Peptides
DNA-Binding Proteins
Recombinant Proteins
Rats
Amino Acids
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Studies of the domain structure of mammalian DNA polymerase β. Identification of a discrete template binding domain. / Kumar, A.; Widen, Steven; Williams, K. R.; Kedar, P.; Karpel, R. L.; Wilson, S. H.

In: Journal of Biological Chemistry, Vol. 265, No. 4, 29.08.1990, p. 2124-2131.

Research output: Contribution to journalArticle

Kumar, A. ; Widen, Steven ; Williams, K. R. ; Kedar, P. ; Karpel, R. L. ; Wilson, S. H. / Studies of the domain structure of mammalian DNA polymerase β. Identification of a discrete template binding domain. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 4. pp. 2124-2131.
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