Prof. Alexander Büerkle

Participant code: UKON

Prof. Alexander Büerkle

Molecular Toxicology Group, Department of Biology, Box X911, University of Konstanz, D-78457 Konstanz, Germany.

Group’s experience:

The group is headed by Prof Alexander Buerkle, Chair of Molecular Toxicology, Dept of Biology, University of Konstanz, Germany. Before his appointment in 2002, he has been a Senior Lecturer at the Department of Gerontology, Institute for Ageing and Health, University of Newcastle upon Tyne, UK, and previously he had been a research group leader at the Institute of Applied Tumour Virology, German Cancer Research Centre, Heidelberg, for over a decade. The major focus of the group is the elucidation of the biological functions of poly(ADP-ribosyl)ation [3], in particular with respect to ageing and longevity, using methodology of toxicology, molecular and cell biology and biochemistry. The group could establish a positive correlation of cellular poly(ADP-ribosyl)ation capacity in leukocytes with life span of 13 mammalian species studied and has followed this up recently at the mechanistic level. They could also show significantly higher poly(ADP-ribosyl)ation capacity in lymphoblastoid cell lines derived from centenarians compared to controls, providing further support for the notion that longevity is associated with a high poly(ADP-ribosyl)ation capacity. In extensive work on transfected cell cultures overexpressing either PARP-1 or a dominant negative version thereof, they could demonstrate that poly(ADP-ribosyl)ation is a key regulator of and assurance factor for genomic stability under conditions of genotoxic stress [4], which is very compatible with the above-mentioned association of longevity with higher poly(ADP-ribosyl)ation capacity. At the level of methodology, the group succeeded establishing a non-radioactive immuno-dot-blot assay to quantify cellular poly(ADP-ribosyl)ation capacity [5] as well as an automated version of the classical FADU method to quantify DNA strand breaks in live cells (protected by US patent 6,210,893). These assays will extensively be used the present project. The Chair of Molecular Toxicology, comprising two closely interacting research groups, is equipped with a full range of the most modern infrastructure necessary to perform the work proposed, including containment level II laboratories and state-of-the-art equipment for molecular & cellular biology and biochemistry work.

Personnel involved:

1) Prof Alexander Buerkle, (M)group leader; scientific supervision, responsible for Workpackage 3
2) Dr Sascha Beneke (F) (postdoc); molecular biology and biochemistry of poly(ADP-ribosyl)ation
3) Dr Christine Brabeck (F) (postdoc); biochemistry of poly(ADP-ribosyl)ation and DNA repair
4) Dr Joerg Diefenbach (M) (postdoc); molecular biology of poly(ADP-ribosyl)ation
5) Mr Thilo Sindlinger (M) (technician); immuno-dot-blot assay to measure PARP-1 activity and automated FADU assay to measure DNA strand breaks and repair.
6) Postdoc to be appointed; salary applied for in the present proposal

Recent relevant publications:

1) Alvarez-Gonzalez R, Spring H, Müller M, Bürkle A. Selective loss of poly(ADP-ribose) and the 85-kDa fragment of poly(ADP-ribose) polymerase in nucleoli during alkylation-induced apoptosis of HeLa cells. J Biol Chem 1999, 274, 32122-32126.

2) Pfeiffer R, Brabeck C, Bürkle A. Quantitative nonisotopic immuno-dot-blot method for the assessment of cellular poly(ADP-ribosyl)ation capacity. Anal Biochem 1999, 275, 118-122.

3) Meyer R, Müller M, Beneke S, Küpper JH, Bürkle A. Negative regulation of alkylation-induced sister-chromatid exchanges by poly(ADP-ribose) polymerase-1 activity. Int J Cancer 2000, 88, 351-355.

4) Beneke S, Alvarez-Gonzalez R, Bürkle A. Comparative characterisation of poly(ADP-ribose) polymerase-1 from two mammalian species with different life span. Exp Gerontol 2000, 35, 989-1002.

5) Bürkle A. Physiology and pathophysiology of poly(ADP-ribosyl)ation. BioEssays 2001, 23, 795-806.

6) Bürkle A. PARP-1: a regulator of genomic stability linked with mammalian longevity. ChemBioChem 2001, 2, 725-728.

7) Brown ML, Franco D, Bürkle A, Chang Y, Role of poly(ADP-ribosyl)ation in DNA-PKcs-independent V(D)J recombination. Proc Natl Acad Sci USA 2002, 99, 4532-4537.

8) Hartwig A, Pelzer A, Asmuss M, Bürkle A. Very low concentrations of arsenite suppress poly(ADP-ribosyl)ation in mammalian cells. Int J Cancer 2003, 104, 1-6.

9) Brabeck C, Pfeiffer R, Leake L, Beneke S, Meyer R, Bürkle A. L-Selegiline potentiates cellular poly(ADP-ribose) formation induced by DNA damage. J Pharmacol Exp Ther 2003, 306, 973-979.

10) Beneke S, Diefenbach J, Bürkle A. Poly(ADP-ribosyl)ation inhibitors as therapeutic drug candidates. Int J Cancer 2004, in press.