Nuclear organisation and epigenetic regulation of gene expression in Dictyostelium discoideum

A series of vectors for the over-expression of tagged proteins in Dictyostelium were designed, constructed and tested. These vectors allow the addition of an N- or C-terminal tag (GFP, RFP, 3xFLAG, 3xHA, 6xMYC and TAP) with an optimized polylinker sequence and no additional amino acid residues at the N or C terminus. Different selectable markers (Blasticidin and gentamicin) are available as well as an extra chromosomal version; these allow copy number and thus expression level to be controlled, as well as allowing for more options with regard to complementation, co- and super-transformation. Finally, the vectors share standardized cloning sites, allowing a gene of interest to be easily transfered between the different versions of the vectors as experimental requirements evolve. The organisation and dynamics of the Dictyostelium nucleus during the cell cycle was investigated. The centromeric histone H3 (CenH3) variant serves to target the kinetochore to the centromeres and thus ensures correct chromosome segregation during mitosis and meiosis. A number of Dictyostelium histone H3-domain containing proteins as GFP-tagged fusions were expressed and it was found that one of them functions as CenH3 in this species. Like CenH3 from some other species, Dictyostelium CenH3 has an extended N-terminal domain with no similarity to any other known proteins. The targeting domain, comprising α-helix 2 and loop 1 of the histone fold is required for targeting CenH3 to centromeres. Compared to the targeting domain of other known and putative CenH3 species, Dictyostelium CenH3 has a shorter loop 1 region. The localisation of a variety of histone modifications and histone modifying enzymes was examined. Using fluorescence in situ hybridisation (FISH) and CenH3 chromatin-immunoprecipitation (ChIP) it was shown that the six telocentric centromeres contain all of the DIRS-1 and most of the DDT-A and skipper transposons. During interphase the centromeres remain attached to the centrosome resulting in a single CenH3 cluster which also contains the putative histone H3K9 methyltransferase SuvA, H3K9me3 and HP1 (heterochromatin protein 1). Except for the centromere cluster and a number of small foci at the nuclear periphery opposite the centromeres, the rest of the nucleus is largely devoid of transposons and heterochromatin associated histone modifications. At least some of the small foci correspond to the distal telomeres, suggesting that the chromosomes are organised in a Rabl-like manner. It was found that in contrast to metazoans, loading of CenH3 onto Dictyostelium centromeres occurs in late G2 phase. Transformation of Dictyostelium with vectors carrying the G418 resistance cassette typically results in the vector integrating into the genome in one or a few tandem arrays of approximately a hundred copies. In contrast, plasmids containing a Blasticidin resistance cassette integrate as single or a few copies. The behaviour of transgenes in the nucleus was examined by FISH, and it was found that low copy transgenes show apparently random distribution within the nucleus, while transgenes with more than approximately 10 copies cluster at or immediately adjacent to the centromeres in interphase cells regardless of the actual integration site along the chromosome. During mitosis the transgenes show centromere-like behaviour, and ChIP experiments show that transgenes contain the heterochromatin marker H3K9me2 and the centromeric histone variant H3v1. This clustering, and centromere-like behaviour was not observed on extrachromosomal transgenes, nor on a line where the transgene had integrated into the extrachromosomal rDNA palindrome. This suggests that it is the repetitive nature of the transgenes that causes the centromere-like behaviour. A Dictyostelium homolog of DET1, a protein largely restricted to multicellular eukaryotes where it has a role in developmental regulation was identified. As in other species Dictyostelium DET1 is nuclear localised. In ChIP experiments DET1 was found to bind the promoters of a number of developmentally regulated loci. In contrast to other species where it is an essential protein, loss of DET1 is not lethal in Dictyostelium, although viability is greatly reduced. Loss of DET1 results in delayed and abnormal development with enlarged aggregation territories. Mutant slugs displayed apparent cell type patterning with a bias towards pre-stalk cell types.

[Ressenya del llibre Paysage et système: de l'organisation écologique à l'organisation visuelle]

Ressenya del llibre Paysage et système: de l'organisation écologique à l'organisation visuelle. L’obra és el resultat de la confrontació dels enfocaments d’investigadors de Bèlgica, Canadà, França i Polònia sobre geografia, ecologia, agronomia i arquitectura del paisatge en el marc de la reunió anual de l’Association Canadienne des Géographes celebrat a la Universitat d’Ottawa el juny de 1982

Organisation and time management

University of Southampton, Dyslexia Services have developed a range of academic study skills resources available to download. This resource supports organisation and time management skills.

On the organisation of Educational Development Centres

NETTLE Project Publication

INFO2009 CW2 2012-13 Briefing, advice and organisation

Notes on group organisation process and planning

A contrastive analysis of non-native interlanguage english intonation systems and their implication in the organisation of information from a functional perspectiva : a study based on a computerized corpus of spanish learners of english

El contexto teórico y empírico de esta investigación sobre entonación, se enmarca dentro de la filosofía lingüística de la teoría sistémico-funcional. El modelo metodológico empleado se basa en la Lingüística de Corpus. La descripción de la adquisición y aprendizaje de la lengua extranjera está justificada dentro del marco teórico de la teoría de interlengua y la adquisición de segundas lenguas y lenguas extranjeras.. El presente estudio del corpus comparativo y longitudinal de aprendices y hablantes nativos de lengua inglesa, tiene como objetivo principal investigar los modelos de entonación producidos por ambos grupos de hablantes. Se pretende demostrar que la diferencia a nivel entonativo entre estos dos grupos no sólo tiene como resultado que los no nativos tengan acento extranjero; sino que puede afectar al mensaje transmitido, en cuanto a la estructura y organización de la información dentro del discurso oral en las metafunciones textual e interpersonal.. Se asume la existencia de un sistema entonativo de interlengua, de esta forma este análisis tiene como objeto no sólo reflejar los errores sino también los posibles sistemas aproximativos de los aprendices mencionados..

Impact of social capital on flexible work organisation models : comparative study of european countries. 'El impacto del capital social en modelos flexibles de organización del trabajo: un estudio comparativo de países europeos'.

Resumen basado en el de la publicación

Online learning matters : extending your organisation's learning provision.

El libro explica, desde un punto de vista organizativo, cómo preparar cursos que ofrezcan formación online o semipresencial en el ámbito de la educación de adultos. Incluye: conceptos del aprendizaje, aprendizaje en línea y el papel de las metodologías online para ampliar y mejorar la oferta educativa; diseño del espacio de aprendizaje; desarrollo de la oferta educativa online; facilitar el aprendizaje en un entorno online; recursos y creación de contenidos; garantía de calidad y control.