Dependence of trans-ADP-ribosylation and nuclear glycolysis on the Arg 34–ATP complex of Zn2+ finger I of poly-ADP-ribose polymerase-1

Kun, Ernest; Kirsten, Eva; Hakam, Alaeddin; Bauer, Pal I.; Mendeleyev, Jerome

doi:10.1016/j.febslet.2008.06.052

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Volume 582, Issue 18 , Pages 2709-2713, 6 August 2008

Dependence of trans-ADP-ribosylation and nuclear glycolysis on the Arg 34–ATP complex of Zn²⁺ finger I of poly-ADP-ribose polymerase-1

Edited by Judit Ovádi

Ernest Kun
- Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, CA 94143, United States
- Department of Anatomy, University of California-San Francisco, School of Medicine, San Francisco, CA 94143, United States
- Department of Cellular and Molecular Pharmacology, University of California-San Francisco, School of Medicine, San Francisco, CA 94143, United States
- Corresponding author. Address: Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, CA 94143, United States. Fax: +1 415 388 7024.
Eva Kirsten
- Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, CA 94143, United States
Alaeddin Hakam
- Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, CA 94143, United States
Pal I. Bauer
- Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, CA 94143, United States
- Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary
Jerome Mendeleyev
- Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, CA 94143, United States

Received 19 May 2008; received in revised form 14 June 2008; accepted 27 June 2008. published online 09 July 2008.

Abstract

The H-bonded complex of ATP with Arg 34 of Zn²⁺ finger I of poly-ADP-ribose polymerase-1 (PARP-1) determines trans-oligo-ADP-ribosylation from NAD⁺ to proteins other than PARP-1. This mechanism was tested in lysolecithin fractions of non-malignant and cancer cells separately and after their recombination. Cellular PARP-1 activity was recovered when the centrifugal sediment was recombined with the supernatant fraction containing cellular ADP-ribose oligomer acceptor proteins. Combination of the matrix fraction (Mx) of cancer cells (lacking OXPHOS) with its supernatant had the same PARP-1 activity as the Mx alone.

The supernatant of non-malignant cells was replaced by glycolytic enzymes as ADP-ribose acceptor. The hexokinase activity of the supernatant increased when OXPHOS of intact cells was uncoupled by carbonyl cyanide 4-(trifluoro methoxy) phenylhydrazone. trans-ADP-ribosylation was demonstrated by polyacrylamide gel electrophoresis.

Abbreviations: PARP-1, poly-ADP-ribose polymerase-1, LL, lysolecithin, Mx, matrix fraction, Su, metabolic fraction, AK, adenylate kinase, HK, hexokinase, FCCP, carbonyl cyanide 4-(trifluoro methoxy) phenylhydrazone