Julia Fuchs

Publications (3)10.16 Total impact

• Source

  Available from: Arnaud Buhot

  Article: TOX4 and its binding partners recognize DNA adducts generated by platinum anticancer drugs.

  Christophe Bounaix Morand du Puch, Ewa Barbier, Alexandra Kraut, Yohann Couté, Julia Fuchs, Arnaud Buhot, Thierry Livache, Michel Sève, Alain Favier, Thierry Douki, Didier Gasparutto, Sylvie Sauvaigo, Jean Breton
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  ABSTRACT: Platinating agents are commonly prescribed anticancer drugs damaging DNA. Induced lesions are recognized by a wide range of proteins. These are involved in cellular mechanisms such as DNA repair, mediation of cytotoxicity or chromatin remodeling. They therefore constitute crucial actors to understand pharmacology of these drugs. To expand our knowledge about this subproteome, we developed a ligand fishing trap coupled to high throughput proteomic tools. This trap is made of damaged plasmids attached to magnetic beads, and was exposed to cell nuclear extracts. Retained proteins were identified by nanoHPLC coupled to tandem mass spectrometry. This approach allowed us to establish a list of 38 proteins interacting with DNA adducts generated by cisplatin, oxaliplatin and satraplatin. Some of them were already known interactome members like high mobility group protein 1 (HMGB1) or the human upstream binding factor (hUBF), but we also succeeded in identifying unexpected proteins such as TOX HMG box family member 4 (TOX4), phosphatase 1 nuclear targeting subunit (PNUTS), and WD repeat-containing protein 82 (WDR82), members of a recently discovered complex. Interaction between TOX4 and platinated DNA was subsequently validated by surface plasmon resonance imaging (SPRi). These interactions highlight new cellular responses to DNA damage induced by chemotherapeutic agents.
  Archives of Biochemistry and Biophysics 03/2011; 507(2):296-303. · 2.93 Impact Factor
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  Available from: Arnaud Buhot

  Article: Effects of formamide on the thermal stability of DNA duplexes on biochips.

  Julia Fuchs, Daniela Dell'Atti, Arnaud Buhot, Roberto Calemczuk, Marco Mascini, Thierry Livache
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  ABSTRACT: In molecular biology, formamide (FA) is a commonly used denaturing agent for DNA. Although its influence on DNA duplex stability in solution is well established, little is known about immobilized DNA on microarrays. We measured thermal denaturation curves for oligonucleotides immobilized by two standard protocols: thiol self-assembling and pyrrole electrospotting. A decrease of the DNA denaturation temperature with increasing FA fraction of the solvent was observed on sequences with mutations for both surface chemistries. The average dissociation temperature decrease was found to be -0.58+/-0.05 degrees C/% FA (v/v) independently of grafting chemistry and probe sequence.
  Analytical Biochemistry 09/2009; 397(1):132-4. · 3.00 Impact Factor
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  Available from: Arnaud Buhot

  Article: SPR imaging for label-free multiplexed analyses of DNA N-glycosylase interactions with damaged DNA duplexes.

  Christelle Corne, Jean-Bernard Fiche, Didier Gasparutto, Valérie Cunin, Emmanuel Suraniti, Arnaud Buhot, Julia Fuchs, Roberto Calemczuk, Thierry Livache, Alain Favier
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  ABSTRACT: Base excision repair (BER) is the major mechanism for the correction of damaged nucleobases resulting from the alkylation and oxidation of DNA. The first step in the BER pathway consists of excision of the abnormal base by several specific DNA N-glycosylases. A decrease in BER activity was found to be related to an increased risk of carcinogenesis and aging. To investigate BER activities we set up a new device for DNA repair analysis based on surface plasmon resonance imaging (SPRi). Oligonucleotides bearing an abnormal nucleoside, namely 8-oxo-7,8-dihydro-2'-deoxyguanosine and (5'S)-5',8-cyclopurine-2'-deoxynucleoside, were grafted by a pyrrole electro-copolymerization process on a glass prism coated with a gold layer. The latter label-free DNA sensor chip permits the detection of N-glycosylase/AP-lyase activity as well as the binding of repair proteins to DNA damage without cleavage activity. Thus, the Fapy DNA N-glycosylase (Fpg) protein is shown as expected to bind and then cleave its natural substrate, namely 8-oxo-7,8-dihydro-guanine, together with the resulting abasic site. Using the current SPR imaging-based DNA array we observed an original binding activity of Fpg towards the (5'S)-5',8-cyclodAdenosine residue. These results altogether show that SPR imaging may be used to simultaneously and specifically detect recognition and excision of several damaged DNA nucleobases, and constitutes an interesting technique to screen inhibitors of DNA repair proteins.
  The Analyst 08/2008; 133(8):1036-45. · 4.23 Impact Factor