Poly(ADP-ribose)-mediated post-translational modification of chromatin-associated human topoisomerase I. Inhibitory effects on catalytic activity

U. N. Kasid, B. Halligan, Leroy-Fong Liu, A. Dritschilo, M. Smulson

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Abstract

We have investigated the association of human topoisomerase I with poly(ADP-ribosylation) domains of chromatin and the effects of this modification on the enzyme activity. In vitro poly(ADP-ribosylation) assays demonstrated that this enzyme was one of the major acceptors for this chromatin-dependent post-translational modification. Western blotting procedures using antibody to topoisomerase I indicated that under extensive poly(ADP-ribosylation) conditions, where a majority of poly(ADP-ribose) acceptor molecules form aggregates, the major population of the topoisomerase I associated with chromatin was apparently non-aggregated. The catalytic activity of the topoisomerase I associated with the poly(ADP-ribosylated) chromatin was 3-5-fold inhibited. Additionally, antibody to poly(ADP-ribose) was used to immunofractionate selectively the modified domains of chromatin. Our data suggests the presence of topoisomerase I, both adjacent and distal to the poly(ADP-ribosylated) sites of chromatin. Unmodified and a significant portion of the modified species of enzyme migrated as approximately 100-kDa proteins. However, the modified form of topoisomerase was noted to be catalytically less active as compared to the enzyme bound to the non-poly(ADP-ribosylated) nucleosomes. These results provide evidence, at the cellular level, for the poly(ADP-ribosylation)-mediated regulation of human topoisomerase I and suggest a functional significance for poly(ADP-ribosylation) in topoisomerase-related processes (replication, transcription, and recombination) in eukaryotes.

Original languageEnglish
Pages (from-to)18687-18692
Number of pages6
JournalJournal of Biological Chemistry
Volume264
Issue number31
Publication statusPublished - 1989
Externally publishedYes

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Poly Adenosine Diphosphate Ribose
Type I DNA Topoisomerase
Post Translational Protein Processing
Adenosine Diphosphate
Chromatin
Catalyst activity
Enzymes
Nucleosomes
Antibodies
Enzyme Assays
Enzyme activity
Transcription
Eukaryota
Genetic Recombination
Assays
Western Blotting
Association reactions
Molecules

ASJC Scopus subject areas

  • Biochemistry

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Poly(ADP-ribose)-mediated post-translational modification of chromatin-associated human topoisomerase I. Inhibitory effects on catalytic activity. / Kasid, U. N.; Halligan, B.; Liu, Leroy-Fong; Dritschilo, A.; Smulson, M.

In: Journal of Biological Chemistry, Vol. 264, No. 31, 1989, p. 18687-18692.

Research output: Contribution to journalArticle

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