caputo.sandrine - Publications List

caputo.sandrine@gmail.com

FORMATION
Doctorat (2002-2006)– Ecole Doctorale InterBio – Univ. Paris VI
Responsable : Dr. Bernard Gilquin et Sophie Zinn-Justin (CEA)
DEA (2001-2002)– Méthodes Spectroscopiques – Univ. Paris VI – Option RMN – Mention bien.
Maîtrise et Licence de Chimie (1999-2001)– Université Paris XII (Créteil) – Spécialité Physique – Mention bien.
DEUG Science de la Vie (1997-1999)– Université Paris XII (Créteil) – Option Chimie.

EXPERIENCE PROFESSIONNELLE
Post-doctorante (depuis avril 2007) – CNRS-ICSN et Ecole Polytechnique – Gif sur Yvette et Palaiseau (91, France)
Analyse structurale de la protéine ribosomale S1 de E. coli et ses interactions avec l’ARN
Responsable : Dr François Bontems (Equipe Lallemand)

Chercheur doctorant (janv 2002-oct 2006)– CEA / Département d’Ingénierie et d’Etude des Protéines – Saclay (91, France)
DEA – CEA / DIEP
Analyse structurale de protéines de l’enveloppe nucléaire impliquées dans des pathologies génétiques.
Responsable : Dr. Bernard Gilquin et Sophie Zinn-Justin (CEA)

COMPETENCES
Techniques: RMN (2D et 3D, Relaxation, interaction protéine-protéine/ADN/ARN), Dichroïsme Circulaire, BIACORE, ITC, Cristallogenèse
Biologie moléculaire (mutagenèse dirigée, amplification de plasmide)
Biochimie : Expression, production et purification de protéines recombinantes chez E. coli
Chromatographie (Gel filtration, chromatographie par affinité, chromatographie par échange d’ions)
Gels SDS-PAGE, Western Blot, Gels retards,GST-pulldown
Synthèse et purification d’ARN
Appareillage utilisé : ÄKTApurifier, Gradifrac, Biorad
Modélisation moléculaire
Culture cellulaire (notions)
Informatique: Environnements : UNIX, Linux, Windows
Logiciels de - RMN (Xwinmr, Nmrpipe, Topspin, Felix, Sparky), Sybyl, Pymol,
- bureautique (Word, Excel, Power-Point)


Journal articles

2008	DOI PMID Aliprandi, Sizun, Perez, Mareuil, Caputo, Leroy, Odaert, Laalami, Uzan, Bontems (2008) S1 ribosomal protein functions in translation initiation and ribonuclease RegB activation are mediated by similar RNA/protein interactions. An NMR and SAXS analysis. J Biol Chem Jan Abstract: The ribosomal protein S1, in Escherichia coli, is necessary for the recognition by the ribosome of the translation initiation codon of most messenger RNAs. It also participates in other functions. In particular, it stimulates the T4 endoribonuclease RegB, which inactivates some of the phage mRNAs, when their translation is no longer required, by cleaving them in the middle of their Shine- Dalgarno sequence. In each function, S1 seems to target very different RNAs, which led to the hypothesis that it possesses different RNA binding sites. We previously demonstrated that the ability of S1 to activate RegB is carried by a fragment of the protein formed of three consecutive domains (domains D3, D4 and D5). The same fragment plays a central role in all other functions. We analyzed its structural organization and its interactions with three RNAs: two RegB substrates and a translation initiation region. We show that these three RNAs bind the same area of the protein through a set of systematic (common to the three RNAs) and specific (RNA-dependent) interactions. We also show that, in the absence of RNA, the D4 and D5 domains are associated while the D3 and D4 domains are in equilibrium between an open (non-interacting) and a closed (weakly interacting) form and that RNA binding induces a structural reorganization of the fragment. All these results suggest that the ability of S1 to recognize different RNAs results from a high adaptability of both its structure and its binding surface. Notes:
2006	DOI PMID Sandrine Caputo, Joël Couprie, Isabelle Duband-Goulet, Emilie Kondé, Feng Lin, Sandrine Braud, Muriel Gondry, Bernard Gilquin, Howard J Worman, Sophie Zinn-Justin (2006) The carboxyl-terminal nucleoplasmic region of MAN1 exhibits a DNA binding winged helix domain. J Biol Chem 281: 26. 18208-18215 Jun Abstract: MAN1 is an integral protein of the inner nuclear membrane that interacts with nuclear lamins and emerin, thus playing a role in nuclear organization. It also binds to chromatin-associated proteins and transcriptional regulators, including the R-Smads, Smad1, Smad2, and Smad3. Mutations in the human gene encoding MAN1 cause sclerosing bone dysplasias, which sometimes have associated skin abnormalities. At the molecular level, these mutations lead to loss of the MAN1-R-Smads interaction, thus perturbing transforming growth factor beta superfamily signaling pathway. As a first step to understanding the physical basis of MAN1 interaction with R-Smads, we here report the structural characterization of the carboxyl-terminal nucleoplasmic region of MAN1, which is responsible for Smad binding. This region exhibits an amino-terminal globular domain adopting a winged helix fold, as found in several Smad-associated sequence-specific DNA binding factors. Consistently, it binds to DNA through the positively charged recognition helix H3 of its winged helix motif. However, it does not show the predicted carboxyl-terminal U2AF homology domain in solution, suggesting that the folding and stability of such a domain in MAN1 depend upon binding to an unidentified partner. Modeling the complex between DNA and the winged helix domain shows that the regions involved in DNA binding are essentially distinct from those reported to be involved in Smad binding. This suggests that MAN1 binds simultaneously to R-Smads and their targeted DNA sequences. Notes:
	DOI PMID Sandrine Caputo, Feng Lin, Bernard Gilquin, Howard J Worman, Sophie Zinn-Justin, Joël Couprie (2006) NMR assignment of region 655-775 of human MAN1. J Biomol NMR 36 Suppl 1: Abstract: Notes:

Journal articles