Objective This project is aimed to study in detail the molecular functions of a group of functionally related evolutionary conservative metazoan transcription factors described by the authors of the project, e.g.(i) TBP-like factor, TRF2, chromatin associated transcription factors ;(ii) e(y)2;(iii) its homologue de(y)2h and;(iv) e(y)3.According to preliminary genetic and molecular data these factors are involved in the control of transcription of many genes. Cloning and primary characterization of e(y)2, de(y)2h and e(y)3 was done in the course of previous INTAS (94-3801) involving almost the same group of authors.The TRF2 gene of Drosophila belongs to a novel family of TATA binding protein (TBP)-like factors. A P-element insertion in the regulatory region of the TRF2 gene induces lawc mutation. Its combination with the phP1 allele enhancing the mutated phenotype induces a more than five times decrease of TRF2 transcription leading to a reduction of expression of some Drosophila genes.In this project(i) a search for genes whose transcription depends on TRF2 will be performed by micro array technique using the RNAs from wild-type strain and from lawc phP1 strain. The nature of promoters belonging to sensitive and resistant genes will be analysed;(ii) search for genes that are modifiers of lawc/TRF2 will be undertaken using the genetic approach based on testing of the collection of genomic deficiencies.The e(y)2 gene present in higher eucaryotes encodes a small (101 amino acid) evolutionary conserved protein, containing no known motifs, which is present in nuclei of all tissues and activates transcription on chromatin templates. It is a component of about 700kDa multi-protein complex also containing TAFII40.It is planned to determine the nature of other proteins forming the e(y)2-containing complex(es) in human and Drosophila using(i) isolation of the complex and analysis its components by MALDI method;(ii) yeast two-hybrid system and;(iii) genetic screen for modifiers of the of e(y)2 mutation in Drosophila.The second gene homologous to e(y)2 (de(y)2h) was found only in Drosophila. As it is expressed only in males, it may be involved in dosage compensation.This hypothesis will be checked(i) by the analysis of de(y)2h protein on polytene chromosomes;(ii) by the analysis of the dosage compensation complex for the presence of de(y)2h;(iii) by the study of proteins interacting with de(y)2h in yeast two hybrid system and;(iv) in experiments on the knock out of either the e(y)2 or de(y)2h gene.The e(y)3 gene encodes a novel 230kD protein containing two DR domains as well as several other known motifs and exists as a part of a large complex with the approximate molecular mass of 1 MD. It has been suggested that it participates in position effect variegation (PEV).The e(y)3 gene has a structural homologue in human. To clarify the molecular role of e(y)3 gene(i) it will be studied whether e(y)3 interacts with other proteins involved in PEV using genetic and molecular methods (in situ colocolization and immuno-precipitation;(ii) a genetic analysis including the search for e(y)3 null mutation and the search for e(y)3 mutation modifiers will be performed. It will also be studied;(iii) how e(y)3 affects the expression of the same gene located either in euchromatin or in heterochromatin. Programme(s) IC-INTAS - International Association for the promotion of cooperation with scientists from the independent states of the former Soviet Union (INTAS), 1993- Topic(s) 4 - Life Sciences OPEN - OPEN Call Call for proposal Data not available Funding Scheme Data not available Coordinator Institut de Génétique et de Biologie Moléculaire et Cellulaire EU contribution No data Address Parc d'Innovation, Rue Laurent Fries 1 67404 Illkirch Cedex France See on map Total cost No data Participants (4) Sort alphabetically Sort by EU Contribution Expand all Collapse all Adolf-Butenandt-Institut-Molekularbiologie Germany EU contribution No data Address Schillerstrasse 44 80336 München See on map Total cost No data Institute of Molecular Biology Russia EU contribution No data Address Vavilov street 32 117334 Moscow See on map Total cost No data Russian Academy of Sciences Russia EU contribution No data Address Vavilov street 34/5 117334 Moscow See on map Total cost No data Siberian branch of Russian Academy of Sciences Russia EU contribution No data Address Lavrentiev prospect, 10 630090 Novosibirsk See on map Total cost No data
