Abstract
We have purified three chromatographically distinct human enzyme activities from HeLa cells, that are capable of converting bleomycin-treated DNA into a substrate for E. coli DNA polymerase I. The bleomycin-treated DNA substrate used in this study has been characterized via a 32P-postlabeling assay and shown to contain strand breaks with 3′-phosphoglycolate termini as greater than 95% of the detectable dose-dependent lesions. The purified HeLa cell enzymes were shown to be capable of removing 3′-phosphoglycolates from this substrate. Also 3′-phosphoglycolate removal and nucleotide incorporation were enzyme dependent. In addition, all three Hela cell enzymes have been determined to possess Class II AP endonuclease activity. The enzymes lack 3′ → 5′ exonuclease activity and are, therefore, dissimilar to exonuclease III - an E. coli enzyme that can remove 3′-phosphoglycolate.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2573-2580 |
| Number of pages | 8 |
| Journal | Nucleic Acids Research |
| Volume | 20 |
| Issue number | 10 |
| State | Published - May 25 1992 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Genetics
- Statistics, Probability and Uncertainty
- Applied Mathematics
- Health, Toxicology and Mutagenesis
- Toxicology
- Genetics(clinical)
Cite this
Human HeLa cell enzymes that remove phosphoglycolate 3′-end groups from DNA. / Winters, Thomas A.; Weinfeld, Michael; Jorgensen, Timothy J.
In: Nucleic Acids Research, Vol. 20, No. 10, 25.05.1992, p. 2573-2580.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Human HeLa cell enzymes that remove phosphoglycolate 3′-end groups from DNA
AU - Winters, Thomas A.
AU - Weinfeld, Michael
AU - Jorgensen, Timothy J.
PY - 1992/5/25
Y1 - 1992/5/25
N2 - We have purified three chromatographically distinct human enzyme activities from HeLa cells, that are capable of converting bleomycin-treated DNA into a substrate for E. coli DNA polymerase I. The bleomycin-treated DNA substrate used in this study has been characterized via a 32P-postlabeling assay and shown to contain strand breaks with 3′-phosphoglycolate termini as greater than 95% of the detectable dose-dependent lesions. The purified HeLa cell enzymes were shown to be capable of removing 3′-phosphoglycolates from this substrate. Also 3′-phosphoglycolate removal and nucleotide incorporation were enzyme dependent. In addition, all three Hela cell enzymes have been determined to possess Class II AP endonuclease activity. The enzymes lack 3′ → 5′ exonuclease activity and are, therefore, dissimilar to exonuclease III - an E. coli enzyme that can remove 3′-phosphoglycolate.
AB - We have purified three chromatographically distinct human enzyme activities from HeLa cells, that are capable of converting bleomycin-treated DNA into a substrate for E. coli DNA polymerase I. The bleomycin-treated DNA substrate used in this study has been characterized via a 32P-postlabeling assay and shown to contain strand breaks with 3′-phosphoglycolate termini as greater than 95% of the detectable dose-dependent lesions. The purified HeLa cell enzymes were shown to be capable of removing 3′-phosphoglycolates from this substrate. Also 3′-phosphoglycolate removal and nucleotide incorporation were enzyme dependent. In addition, all three Hela cell enzymes have been determined to possess Class II AP endonuclease activity. The enzymes lack 3′ → 5′ exonuclease activity and are, therefore, dissimilar to exonuclease III - an E. coli enzyme that can remove 3′-phosphoglycolate.
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M3 - Article
C2 - 1375993
AN - SCOPUS:0026554433
VL - 20
SP - 2573
EP - 2580
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 1362-4962
IS - 10
ER -