eprintid: 1874
rev_number: 7
eprint_status: archive
userid: 1
dir: disk0/00/00/18/74
datestamp: 2002-01-30 13:55:00
lastmod: 2014-01-13 14:15:00
status_changed: 2012-08-14 15:03:06
type: doctoralThesis
metadata_visibility: show
creators_name: Pfeiffer, Ragen
title: Development of methods to investigate poly(ADP-ribose) polymerase-1 activity and DNA base-excision repair in relation to cancer and ageing
title_en: Development of methods to investigate oly(ADP-ribose) polymerase-1 activity and DNA base-excision repair in relation to cancer and ageing
title_de: Entwicklung von Methoden um Poly(ADP-ribose) polymerase-1-aktivität und DNA Basenexcisionsreperatur in Beziehung zu Krebs und Alterung zu untersuchen
ispublished: pub
subjects: 610
divisions: 850300
adv_faculty: af-14
keywords: DNS Schädigung , ToxikologieAgeing , Cancer , DNA repair , DNA damage , Toxicology
cterms_swd: Alterung
cterms_swd: Krebs
cterms_swd: DNS Reparatur
note: Teile in: Anal. Biochem.275, 118-122
abstract: Summary The work presented here comprises the development of a new immuno-dot-blot  method to detect poly(ADP-ribosyl)ation capacity in cells and the automation  of the fluorescence detected alkaline DNA unwinding (FADU) assay to measure  DNA strand breaks and repair.  Poly(ADP-ribose) polymerase-1 (PARP-1) is involved in DNA base-excision repair  and the maintenance of genomic stability, but emerges to have many more  functions in cellular physiology and pathophysiology. Cellular  poly(ADP-ribosyl)ation capacity is correlated with maximal life span in  mammalian species and a with longevity in humans. In this work a new, non-isotopic, simple and reliable method for the assessment  of poly(ADP-ribosyl)ation capacity is presented. In experiments comparing  fibroblasts from wild-type mice (PARP-1+/+) with those from PARP-1 knockout  mice (PARP-1-/-) we could demonstrate that the p(ADPr) formation assessed in  this assay is due almost exclusively to PARP-1 and only in trace amounts to  PARP-2 or other members of the PARP enzyme family.  Here an automated simplified version of the FADU assay is described, that  posesses very high reproducibility. To investigate if DNA base-excision repair capacity or half-life time can be  correlated with the life span of mammals, a preliminary set of comparative  experiments on g-irradiated human and rat blood cells was performed. The  percentage of lesions repaired during the first 50 min after irradiation was  significantly lower in the rat cells (17 vs 45 ). These data suggest that  there might exist a correlation between DNA base excision repair and  mammalian longevity. We have also performed experiments comparing DNA base excision repair in  PARP-1-/- and in PARP-1+/+ fibroblasts. DNA repair was found to be dramatically  impaired in PARP-1-/- cells but very active in PARP-1+/+ cells. These results  represent additional clear evidence for the involvement of PARP-1  in base excision repair. 
abstract_translated_text: Summary The work presented here comprises the development of a new immuno-dot-blot  method to detect poly(ADP-ribosyl)ation capacity in cells and the automation  of the fluorescence detected alkaline DNA unwinding (FADU) assay to measure  DNA strand breaks and repair.  Poly(ADP-ribose) polymerase-1 (PARP-1) is involved in DNA base-excision repair  and the maintenance of genomic stability, but emerges to have many more  functions in cellular physiology and pathophysiology. Cellular  poly(ADP-ribosyl)ation capacity is correlated with maximal life span in  mammalian species and a with longevity in humans. In this work a new, non-isotopic, simple and reliable method for the assessment  of poly(ADP-ribosyl)ation capacity is presented. In experiments comparing  fibroblasts from wild-type mice (PARP-1+/+) with those from PARP-1 knockout  mice (PARP-1-/-) we could demonstrate that the p(ADPr) formation assessed in  this assay is due almost exclusively to PARP-1 and only in trace amounts to  PARP-2 or other members of the PARP enzyme family.  Here an automated simplified version of the FADU assay is described, that  posesses very high reproducibility. To investigate if DNA base-excision repair capacity or half-life time can be  correlated with the life span of mammals, a preliminary set of comparative  experiments on g-irradiated human and rat blood cells was performed. The  percentage of lesions repaired during the first 50 min after irradiation was  significantly lower in the rat cells (17 vs 45 ). These data suggest that  there might exist a correlation between DNA base excision repair and  mammalian longevity. We have also performed experiments comparing DNA base excision repair in  PARP-1-/- and in PARP-1+/+ fibroblasts . DNA repair was found to be dramatically  impaired in PARP-1-/- cells but very active in PARP-1+/+ cells. These results  represent additional clear evidence for the involvement of PARP-1  in base excision repair. 
abstract_translated_lang: eng
date: 2001
date_type: published
id_scheme: DOI
id_number: 10.11588/heidok.00001874
ppn_swb: 1643278738
own_urn: urn:nbn:de:bsz:16-heidok-18742
date_accepted: 2001-12-17
advisor: HASH(0x564e1c608f88)
language: eng
bibsort: PFEIFFERRADEVELOPMEN2001
full_text_status: public
citation:   Pfeiffer, Ragen  (2001) Development of methods to investigate poly(ADP-ribose) polymerase-1 activity and DNA base-excision repair in relation to cancer and ageing.  [Dissertation]     
document_url: https://archiv.ub.uni-heidelberg.de/volltextserver/1874/1/Ragen.pdf