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 language | English (US) |
|---|---|
| Pages (from-to) | 2124-2131 |
| Number of pages | 8 |
| Journal | Journal of Biological Chemistry |
| Volume | 265 |
| Issue number | 4 |
| State | Published - Aug 29 1990 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Studies of the domain structure of mammalian DNA polymerase β. Identification of a discrete template binding domain
AU - Kumar, A.
AU - Widen, Steven
AU - Williams, K. R.
AU - Kedar, P.
AU - Karpel, R. L.
AU - Wilson, S. H.
PY - 1990/8/29
Y1 - 1990/8/29
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0025088519&partnerID=8YFLogxK
M3 - Article
C2 - 2404980
AN - SCOPUS:0025088519
VL - 265
SP - 2124
EP - 2131
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 4
ER -