New polymorphisms in the human poly(ADP-ribose) polymerase-1 coding sequence: lack of association with longevity or with increased cellular poly(ADP-ribosyl)ation capacity

Received: 27 March 2000 / Accepted: 9 July 2000 / Published online: 30 August 2000 © Springer-Verlag 2000

FLORENCE COTTET MARIE-LAURE MUIRAS was a graduate student at the received her Ph.D. from the Ecole Pratique des Hautes University Paris VI, France, in Etudes (EPHE, Paris, France) the field of genetics of human in collaboration with the Fond- longevity. She is presently a ation Jean Dausset-Centre postdoctoral fellow at the Ger- d’étude du Polymorphisme hu- man Cancer Research Centre main (Paris, France) where she (Heidelberg, Germany). Her was interested in the genetics research interests include of human longevity. She is poly(ADP-ribosyl)ation and its now in charge of clinical trials relation to genomic instability    

Abstract Poly(ADP-ribose) polymerase-1 (PARP-1) en- coded by the PARP-1 gene, is a ubiquitous and abundant DNA-binding protein involved in the cellular response to various genotoxic agents. In a previous study we showed that maximal oligonucleotide-stimulated poly(ADP-ribo- syl)ation was significantly higher in permeabilised lym- phoblastoid cell lines from a French population of cente- narians compared with controls aged 20–70 years, sup- porting the notion that longevity is associated with a ge- netically determined, high poly(ADP-ribosyl)ation capaci- ty. Here, we describe four new genetic polymorphisms, three of which represent silent nucleotide variants (C T, 402 T C, G A), and one of which leads to a valine -to- 1011 1215 762 alanine exchange (T C). We undertook an association 2444 study between two of these polymorphisms and human longevity or poly(ADP-ribosyl)ation capacity in permea- bilised lymphoblastoid cells. By analysing 648 DNA sam- ples from a French population (324 centenarians and 324 controls) by fluorescent-allele-specific PCR, we showed the absence of any significant enrichment of any of the ge- notypes in the study of centenarians versus controls. Fur- thermore, we studied genotype distributions from individ- uals who had previously been tested for poly(ADP-ribo- syl)ation capacity. None of the genotype combinations at any polymorphic site studied could be related to a high or low level of poly(ADP-ribosyl)ation capacity. Together, these results strongly suggest that the longevity-related differences in the poly(ADP-ribosyl)ation capacity of hu- man lymphoblastoid cell lines cannot be explained by ge- netic polymorphisms in the PARP-1 coding sequence and that other mechanisms have to be considered as potential

MARIE-LAURE MUIRAS received her Ph.D. from the University Paris VI, France, in the field of genetics of human longevity. She is presently a postdoctoral fellow at the Ger- man Cancer Research Centre (Heidelberg, Germany). Her research interests include poly(ADP-ribosyl)ation and its

was a graduate student at the Ecole Pratique des Hautes Etudes (EPHE, Paris, France) in collaboration with the Fond- ation Jean Dausset-Centre d’étude du Polymorphisme hu- main (Paris, France) where she was interested in the genetics of human longevity. She is now in charge of clinical trials in a pharmaceutical company.

Fondation Jean Dausset-Centre d’Etude du Polymorphisme Humain, 27, rue Juliette Dodu, 75010 Paris, France

Key words Human longevity · Poly(ADP-ribose) polymerase · Centenarian · Allele-specific amplification Genetic polymorphism

A. Bürkle, University of Newcastle, Department of Gerontology, Institute for the Health of the Elderly, Wolfson Research Centre, Newcastle General Hospital, Westgate Road, Newcastle upon Tyne, NE4 6BE, UK

Abbreviations PARP: Poly(ADP-ribose) polymerase · ROS: reactive oxygen species · EBV: Epstein-Barr virus

Introduction

er types of DNA damage, namely poly(ADP-ribosyl)at- ion. This post-translational modification of nuclear pro- teins which facilitates the cellular recovery from DNA damage is catalysed by the nuclear enzyme poly(ADP- ribose) polymerase-1 (PARP-1; EC 2.4.2.30), with NAD+ serving as substrate [16, 17,18]. This enzyme is found in most eukaryotic species and accounts for about 85% of cellular poly(ADP-ribose) formation in mouse em- bryo fibroblasts [19]. Very recently, additional polypep- tides catalysing the poly(ADP-ribosyl)ation reaction have been described [20,21]; however, information re- garding the expression, structure and function of these novel PARPs is still limited. The amino-terminal do- main of PARP-1 binds to single- or double-stranded breaks in DNA. Binding to DNA breaks causes a drastic activation of the catalytic centre in the carboxy-terminal NAD+-binding domain. A number of “acceptor” pro- teins covalently modified with poly(ADP-ribose) have been identified in vivo and in vitro, including histones and PARP-1 itself. Treatment of cells with certain chemical or physical DNA-damaging agents, including γ ROS, alkylating agents, cisplatin [22], and - or ultravi- olet radiation [23], induces a dose-dependent stimula- tion of poly(ADP-ribose) synthesis.