Quantitative correlation between cellular proliferation and nuclear poly (ADP-ribose) polymerase (PARP-1)

Ernest Kun, Eva Kirsten, P. Bauer, Charles P. Ordahl

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Treatment of cells with lysophosphatidyl choline and centrifugal extraction can separate poly (ADP-ribose) synthetase (PARP-1) and DNA synthetase activities, permitting the experimental analysis and comparison of both multienzyme systems. Only PARP-1 is being assayed by our system. Ca2+ and Mg2+ have minor activating effects, and added histones are without activating action. Short end-blocked dsDNAs at nM concentrations and spermine at mM concentrations are maximally activating coenzymes of poly (ADP-ribose) synthesis. Comparison of non-proliferating non-malignant cells with rapidly growing cancer cells demonstrates that rates of poly (ADP-ribose) synthesis and DNA synthesis are highest in pre-confluent non-malignant cells and in proliferating cancer cells, and lowest in contact-inhibited non-malignant cells. Rates of poly (ADP-ribose) synthesis correlate with the number of enzymatically activable PARP-1 molecules per cell, determined under Vmax conditions where activity is linearly proportional to enzyme protein. Contact-inhibited non-malignant cells exhibit only trans-ADP-ribosylation that is not affected by ATP, while rapid growth, especially in cancer cells, demonstrates extensive auto-poly (ADP)-ribosylation that is strongly inhibited by ATP at concentrations present in cells exhibiting normal bioenergetics. Rates of mRNA synthesis in non-proliferating nonmalignant cells and in cancer cells were indistinguishable, indicating that the differences observed between cellular phenotypes are most likely due to reassembly of PARP-1 molecules in nuclei to homo-dimers (in cancer cells) and hetero-dimers (in non-cancer cells). A specific inhibitor and an inactivator of PARP-1 each inhibit DNA synthesis when intact cancer cells are pretreated with these drugs. Direct addition of these drugs to permeabilized cells performing DNA synthesis has no effect on DNA synthesis. The most striking diagnostic signal for cancer cells is activation of PARP-1 and of DNA synthesis.

Original languageEnglish
Pages (from-to)293-300
Number of pages8
JournalInternational Journal of Molecular Medicine
Volume17
Issue number2
Publication statusPublished - Feb 2006

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Cell Proliferation
Poly Adenosine Diphosphate Ribose
DNA
Neoplasms
Poly (ADP-Ribose) Polymerase-1
Ligases
Adenosine Diphosphate
Adenosine Triphosphate
Spermine
Coenzymes
Choline
Pharmaceutical Preparations
Histones
Energy Metabolism
Phenotype

Keywords

  • AMP-PNP
  • Control of DNA synthesis by PARP-1 activity
  • Lysophosphatidyl choline permeabilized cell fractionation of PARP-1 and DNA synthetase systems
  • PARP-1 mRNA analysis

ASJC Scopus subject areas

  • Genetics

Cite this

Quantitative correlation between cellular proliferation and nuclear poly (ADP-ribose) polymerase (PARP-1). / Kun, Ernest; Kirsten, Eva; Bauer, P.; Ordahl, Charles P.

In: International Journal of Molecular Medicine, Vol. 17, No. 2, 02.2006, p. 293-300.

Research output: Contribution to journalArticle

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