Human HeLa cell enzymes that remove phosphoglycolate 3′-end groups from DNA

Thomas A. Winters, Michael Weinfeld, Timothy J. Jorgensen

Research output: Contribution to journalArticle

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 languageEnglish (US)
Pages (from-to)2573-2580
Number of pages8
JournalNucleic Acids Research
Volume20
Issue number10
StatePublished - May 25 1992
Externally publishedYes

Fingerprint

HeLa Cells
Enzymes
DNA
Cell
Bleomycin
Escherichia coli
Substrate
Substrates
Escherichia Coli
DNA-(Apurinic or Apyrimidinic Site) Lyase
DNA Polymerase I
Exonucleases
Enzyme activity
Nucleotides
Dependent
Assays
Human Activities
phosphoglycolate
Human
Dose

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Winters, T. A., Weinfeld, M., & Jorgensen, T. J. (1992). Human HeLa cell enzymes that remove phosphoglycolate 3′-end groups from DNA. Nucleic Acids Research, 20(10), 2573-2580.

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 journalArticle

Winters, TA, Weinfeld, M & Jorgensen, TJ 1992, 'Human HeLa cell enzymes that remove phosphoglycolate 3′-end groups from DNA', Nucleic Acids Research, vol. 20, no. 10, pp. 2573-2580.
Winters TA, Weinfeld M, Jorgensen TJ. Human HeLa cell enzymes that remove phosphoglycolate 3′-end groups from DNA. Nucleic Acids Research. 1992 May 25;20(10):2573-2580.
Winters, Thomas A. ; Weinfeld, Michael ; Jorgensen, Timothy J. / Human HeLa cell enzymes that remove phosphoglycolate 3′-end groups from DNA. In: Nucleic Acids Research. 1992 ; Vol. 20, No. 10. pp. 2573-2580.
@article{a87f8c476644429c96906195a52444c2,
title = "Human HeLa cell enzymes that remove phosphoglycolate 3′-end groups from DNA",
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.",
author = "Winters, {Thomas A.} and Michael Weinfeld and Jorgensen, {Timothy J.}",
year = "1992",
month = "5",
day = "25",
language = "English (US)",
volume = "20",
pages = "2573--2580",
journal = "Nucleic Acids Research",
issn = "1362-4962",
publisher = "Oxford University Press",
number = "10",

}

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.

UR - http://www.scopus.com/inward/record.url?scp=0026554433&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026554433&partnerID=8YFLogxK

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 -