Evaluation de l'utilisation du portfolio par les internes en médecine générale d'Angers

Contexte - Le portfolio est un outil de validation reconnu du parcours des internes en médecine générale. Articulé autour des activités d apprentissage, d enseignement et d évaluation, il permet l acquisition d une démarche réflexive et une évaluation formative. Il implique également la mise en œuvre du feedback pédagogique par le biais du tutorat et l analyse des Récits de Situation Complexe Authentique (RSCA). Objectifs - Les internes qui ont validé leur troisième année de Diplôme d Etudes Spécialisées (DES) de médecine générale à la faculté d Angers en 2010 constituent la première promotion angevine à avoir utilisé le portfolio dès le début de leur formation. Cette étude réalise un premier état des lieux afin d optimiser l utilisation future de cet outil. Méthode - Un questionnaire d évaluation, dont les items ont été ciblés par un focus group, a été soumis à tous les internes angevins inscrits en 3ème année de DES de médecine générale pour l année universitaire 2009-2010. Résultats - La majorité reconnaît l intérêt du portfolio dans l analyse des pratiques professionnelles. Le nouveau rôle actif de l apprenant est bien intégré mais reste dépendant de la qualité de la relation avec le tuteur. Le frein principal à la constitution du portfolio est le manque de temps. La crainte de s exposer à travers les RSCA est également évoquée. Conclusion - Une utilisation optimale du portfolio nécessite une présentation claire en début de DES puis l investissement réciproque de la part de l interne et du tuteur tout au long du cursus. Le processus d intégration de cet outil doit être accéléré en première année pour éviter une surcharge de travail en fin de DES.ANGERS-BU Médecine-Pharmacie (490072105) / SudocSudocFranceF

Multiple Roles of the τ131 Subunit of Yeast Transcription Factor IIIC (TFIIIC) in TFIIIB Assembly

Yeast transcription factor IIIC (TFIIIC) plays a key role in assembling the transcription initiation factor TFIIIB on class III genes after TFIIIC-DNA binding. The second largest subunit of TFIIIC, τ131, is thought to initiate TFIIIB assembly by interacting with Brf1/TFIIIB70. In this work, we have analyzed a TFIIIC mutant (τ131-ΔTPR2) harboring a deletion in τ131 removing the second of its 11 tetratricopeptide repeats. Remarkably, this thermosensitive mutation was selectively suppressed in vivo by overexpression of B”/TFIIIB90, but not Brf1 or TATA-binding protein. In vitro, the mutant factor preincubated at restrictive temperature bound DNA efficiently but lost transcription factor activity. The in vitro transcription defect was abolished at high concentrations of B” but not Brf1. Copurification experiments of baculovirus-expressed proteins confirmed a direct physical interaction between τ131 and B”. τ131, therefore, appears to be involved in the recruitment of both Brf1 and B”

Identification of proteins associated with RNA polymerase III using a modified tandem chromatin affinity purification.

International audienceTo identify the proteins associated with the RNA polymerase III (Pol III) machinery in exponentially growing yeast cells, we developed our own tandem chromatin affinity purification procedure (TChAP) after in vivo cross-link, allowing a reproducible and good recovery of the protein bait and its associated partners. In contrast to TFIIIA that could only be purified as a free protein, this protocol allows us to capture free Pol III together with Pol III bound on its target genes. Transcription factors, elongation factors, RNA-associated proteins and proteins involved in Pol III biogenesis were identified by mass spectrometry. Interestingly, the presence of all the TFIIIB subunits found associated with Pol III together with the absence of TFIIIC and chromatin factors including histones suggest that DNA-bound Pol III purified using TChAP is mainly engaged in transcription reinitiation

EWS, but Not EWS-FLI-1, Is Associated with Both TFIID and RNA Polymerase II: Interactions between Two Members of the TET Family, EWS and hTAF(II)68, and Subunits of TFIID and RNA Polymerase II Complexes

The t(11;22) chromosomal translocation specifically linked to Ewing sarcoma and primitive neuroectodermal tumor results in a chimeric molecule fusing the amino-terminus-encoding region of the EWS gene to the carboxyl-terminal DNA-binding domain encoded by the FLI-1 gene. As the function of the protein encoded by the EWS gene remains unknown, we investigated the putative role of EWS in RNA polymerase II (Pol II) transcription by comparing its activity with that of its structural homolog, hTAF(II)68. We demonstrate that a portion of EWS is able to associate with the basal transcription factor TFIID, which is composed of the TATA-binding protein (TBP) and TBP-associated factors (TAF(II)s). In vitro binding studies revealed that both EWS and hTAF(II)68 interact with the same TFIID subunits, suggesting that the presence of EWS and that of hTAF(II)68 in the same TFIID complex may be mutually exclusive. Moreover, EWS is not exclusively associated with TFIID but, similarly to hTAF(II)68, is also associated with the Pol II complex. The subunits of Pol II that interact with EWS and hTAF(II)68 have been identified, confirming the association with the polymerase. In contrast to EWS, the tumorigenic EWS–FLI-1 fusion protein is not associated with either TFIID or Pol II in Ewing cell nuclear extracts. These observations suggest that EWS and EWS–FLI-1 may play different roles in Pol II transcription

Dicistronic tRNA-5S rRNA genes in Yarrowia lipolytica: an alternative TFIIIA-independent way for expression of 5S rRNA genes.

International audienceIn eukaryotes, genes transcribed by RNA polymerase III (Pol III) carry their own internal promoters and as such, are transcribed as individual units. Indeed, a very few cases of dicistronic Pol III genes are yet known. In contrast to other hemiascomycetes, 5S rRNA genes of Yarrowia lipolytica are not embedded into the tandemly repeated rDNA units, but appear scattered throughout the genome. We report here an unprecedented genomic organization: 48 over the 108 copies of the 5S rRNA genes are located 3' of tRNA genes. We show that these peculiar tRNA-5S rRNA dicistronic genes are expressed in vitro and in vivo as Pol III transcriptional fusions without the need of the 5S rRNA gene-specific factor TFIIIA, the deletion of which displays a viable phenotype. We also report the existence of a novel putative non-coding Pol III RNA of unknown function about 70 nucleotide-long (RUF70), the 13 genes of which are devoid of internal Pol III promoters and located 3' of the 13 copies of the tDNA-Trp (CCA). All genes embedded in the various dicistronic genes, fused 5S rRNA genes, RUF70 genes and their leader tRNA genes appear to be efficiently transcribed and their products correctly processed in vivo