ETUDE DES INTERACTIONS ENTRE PROTEINES ET LESIONS DE L'ADN PAR RESONANCE PLASMONIQUE DE SURFACE PAR IMAGERIE (SPRI)

Abstract : DNA is the carrier of genetic information. DNA damage caused by various physical or chemical stresses is a challenge for cellular repair systems. These include the base excision repair system (BER) which involves several enzymes whose objectives are the recognition and removal of damaged bases, well-recognised functions for two glycosylases: prokaryotic Fpg and eukaryotic OGG1. Many approaches have been described to study DNA / protein interactions in vitro. With surface plasmon resonance imaging (SPRi), we have a real-time technique, without labeling, with which we can observe interactions in parallel for a single protein purified enzyme (Fpg, OGG1, EndoIV or Ape1) vis-à-vis various injuries to synthetic oligonucleotides immobilized on a gold surface. The damages studied were an oxidized base (8-oxoG), a cyclised base (cycloadenine) and analogues of abasic sites (THF and C3). We also studied the action of these enzymes on multiple lesions, in tandem, combining the 8-oxoG and 8-oxoA bases on the same strand of DNA. The originality of our system combines the direct analysis of the DNA / protein interaction with the indirect approach of observing its outcome by hybridization and amplification of the signal after a thermal ramp. The results obtained enable us to consider the use of our technique to observe the simultaneous repair of certain lesions by cell extracts for biochemical work, or by human tissue extracts for bio-medical work.
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Theses
Other. Université Joseph-Fourier - Grenoble I, 2010. French


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Submitted on : Friday, July 23, 2010 - 11:37:33 AM
Last modification on : Friday, July 23, 2010 - 11:41:33 AM

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Christelle Corne. ETUDE DES INTERACTIONS ENTRE PROTEINES ET LESIONS DE L'ADN PAR RESONANCE PLASMONIQUE DE SURFACE PAR IMAGERIE (SPRI). Other. Université Joseph-Fourier - Grenoble I, 2010. French. <tel-00505298>

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