Published October 3, 2003 | Version v1
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Radiation-induced cross-link DNA damages: synthesis, measurement and insertion into oligonucleotides for replication and enzymatic repair studies

Creators

Description

This research thesis addresses the synthesis, measurement and study of the biological impact of radio-induced DNA double damages. In the first part, the author reports the study of the reactivity and fate of the 5-(2'-desoxy-uridilyl)methyl radical which is one of the intermediates formed by oxidizing photo-sensitisation of thymine. The next part reports results of the formation and measurement of double damages of isolated and cellular DNA, notably in the case of γ irradiation. The third part reports the study of in vitro replication of one of the double damages. The behaviour of different polymerases with respect to the damage is reported. Finally, the modified oligonucleotide has been used as a substrate to highlight possible activities of enzymatic repair for this type of cross-link damages by purified proteins or proteins present within cellular extracts

Abstract (French)

Le travail de these rapporte ici, a ete dedie a la synthese, a la mesure et a l'etude de l'impact biologique de lesions doubles radio-induites de l'ADN. Le chapitre I est consacre a la reactivite et le devenir du radical 5-(2'-desoxyuridilyl)methyle, un des principaux intermediaires formes par photosensibilisation oxydative de la thymine. Un nucleoside modifie precurseur de ce radical, la 5-(phenylthiomethyl)-2'-desoxyuridine (PhSdUrd), a ete synthetise, puis incorpore dans des oligonucleotides. L'intermediaire radicalaire a ete genere dans les chaines oligonucleotidiques par voie photochimique. Les oligonucleotides contenant la lesion pontee entre le methyle exocyclique de la thymine et le C-8 de la purine adjacente, ont ete isoles, puis caracterises par diverses methodes physico-chimiques et enzymologiques. Un oligonucleotide modifie de 21 residus de longueur a ete prepare afin d'etre utilise pour des etudes de replication et de reparation d'un dommage insere; ce dernier est une lesion double qui a ete generee de maniere specifique dans un site determine du fragment d'ADN. Le deuxieme chapitre rapporte les resultats sur la formation et la mesure des lesions doubles dans l'ADN isole et cellulaire. Le taux de formation des lesions dans l'ADN soumis a un rayonnement gamma, a ete determine. La troisieme partie concerne l'etude de la replication in vitro d'une des lesions doubles. Les resultats sur le comportement de differentes polymerases vis-a-vis de la lesion tandem sont presentes. Dans le dernier chapitre de ce memoire, l'oligonucleotide modifie a ete utilise comme substrat pour mettre a jour d'eventuelles activites de reparation enzymatique de ce type de lesions pontees vicinales, par des proteines purifiees ou presentes au sein d'extraits cellulaires

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Additional details

Additional titles

Original title (French)
Lesions pontees radio-induites de l'ADN: synthese, mesure et incorporation dans des oligonucleotides pour l'etude de leur replication et de leur reparation

Identifiers

URL
http://www.iaea.org/inis/Contacts/;

Publishing Information

Imprint Pagination
208 p.
Report number
FRCEA-TH--8721

INIS

Country of Publication
France
Country of Input or Organization
France
INIS RN
48078386
Subject category
S63: RADIATION, THERMAL, AND OTHER ENVIRONMENTAL POLLUTANT EFFECTS ON LIVING ORGANISMS AND BIOLOGICAL MATERIALS; S60: APPLIED LIFE SCIENCES;
Resource subtype / Literary indicator
Thesis
Descriptors DEI
DNA; DNA DAMAGES; DNA HELICASES; DNA POLYMERASES; DNA REPAIR; DNA REPLICATION; ENZYME ACTIVITY; GAMMA RADIATION; METHYL RADICALS; METHYL TRANSFERASES; PHOTOREACTIVATION; PHOTOSENSITIVITY; RADIATION INJURIES; STRUCTURE-ACTIVITY RELATIONSHIPS
Descriptors DEC
ALKYL RADICALS; BIOLOGICAL EFFECTS; BIOLOGICAL RADIATION EFFECTS; BIOLOGICAL RECOVERY; BIOLOGICAL REPAIR; CARBON-GROUP TRANSFERASES; DISEASES; ELECTROMAGNETIC RADIATION; ENZYMES; INJURIES; IONIZING RADIATIONS; NUCLEIC ACID REPLICATION; NUCLEIC ACIDS; NUCLEOTIDYLTRANSFERASES; ORGANIC COMPOUNDS; PHOSPHORUS-GROUP TRANSFERASES; POLYMERASES; PROTEINS; RADIATION EFFECTS; RADIATIONS; RADICALS; REPAIR; SENSITIVITY; TRANSFERASES

Optional Information

Notes
212 refs.; Available from the INIS Liaison Officer for France, see the 'INIS contacts' section of the INIS-NKM website for current contact and E-mail addresses: http://www.iaea.org/inis/Contacts/; Also available from Bibliotheques Universitaires Grenoble Alpes, scd@ujf-grenoble.fr