Objective
The objective of this proposal is to initiate a close collaboration between two INTAS laboratories (J-A. Lepesant, Institut Jacques Monod, Paris, and R. Nöthiger, University of Zurich) and two NIS laboratories (I. Zhimulev, Academy of Sciences, Novosibirsk, and D. Koryakov, University of Novosibirsk) who share a common interest for developmental genetics in the fruitfully Drosophila melanogaster. These laboratories have an extensive experience in cytogenetics, genetics and mollar biology of gene expression in this model organism. They have not formally collaborated in the past, but they have developed a continuous and privileged exchange of scientific information, experimental protocols and biological material. In joining more tightly their efforts, their goal is to make an increased profit of their complementary expertise to investigate in greater detail two genetic systems which modulate, through their influence on the structure of chromatin or transcription complexes, the expression of a large fraction of the Drosophila genome during development. One is the Broad-Complex (BR-C) gene which encodes a family of Zinc finger trans-regulatory proteins which regulate, in response to the steroid hormone ecdysone, the activity of a large set of target genes required for the initiation and completion of metamorphosis. The second are the Sex-lethal and male-specific lethal (msl) genes which, in addition to somatic sexual differentiation, control dosage compensation by hypertranscribing the X-linked genes in males. The programme is composed of two subprojects with distinct specific aims. The aims of subproject 1 on the Broad-Complex (BR-C) are: to isolate and characterize at the genetic and mollar level a large series of novel mutations of the BR-C gene generated by insertion or excision of a transposable element. It is expected that this mutagenesis will firmly establish the relationship between the different genetic functions carried out by the BR-C and the protein isoforms it encodes. It should also provide a powerful approach for the identification and functional dissection of the regulatory elements controling the highly complex transcriptional pattern of the BR-C. To examine puffing activity in salivary gland polytene chromosomes in order to analyse, at the whole genome level, the alterations induced by these mutations on the pattern of transcriptional activity of BR-C target genes. It is expected that this study will deepen the understanding of the role of the BR-C gene in the regulation of the cascade of gene activity triggered by the hormonal signal. The aims of subproject 2 on the Sxl and msl genes are: to examine the structure and transcriptional activity of salivary gland polytene chromosomes in order to investigate the specific and differential alterations of chromatin structure brought about by inactivation or misexpression of these genes in males or in females. It is expected that this study will provide a direct test of the hypothesis that the Sxl and msl genes take part in the genetic control of heterochromatinisation of extended genomic regions which are locked in a permanent state of transcriptional inactivity during development as a result of this process. to examine the alterations induced by the mutation or misexpression of these genes on the extent and timing of replication activity in these heterochromatic regions. This study should provide an assessment of the role of the Sxl and msl genes in the regulation of replication of these particular genomic regions in correlation with their transcriptional activity. It is anticipated that the proposed research project will advance the understanding of chromosome structure and transcriptional regulation, and that the four participating laboratories will technically and intellectually profit from the collaboration. An additional and equally important objective of the proposal is the transfer and implementation in the Russian laboratories of technologies and methods currently used in the INTAS laboratories.
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Programme(s)
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Coordinator
75251 Paris
France
The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.