Obiettivo The project plans a multi-lateral cooperation involving two Russian (Moscow and Pushchino) and two Western (Dutch and Finnish) groups and is aimed at investigation of some fundamental and applied aspects of reproduction and evolution of viruses with positive RNA-strand genome. For 3 of these groups the proposal is a continuation of previous successful INTAS grants (95-1365 and 97-10348). Picornaviruses (polioviruses, coxsackieviruses, parechoviruses) and Q(phage will be used as experimental objects. Multidisciplinary approach including biochemical, biophysical, morphological, and chemical techniques as well as genetic engineering and bioinformatics will be implemented.The following main issues will be addressed(a) The effects of picornavirus infection on the expression of the host cell genome and on the infrastructure of infected cells. To this end, the global mRNA expression in cells infected with different picornaviruses under different conditions will be studied by cDNA microchip technology and attempts to identify key host genes involved in the defensive and pathological reactions will be undertaken. The mechanisms of alteration of intracellular trafficking (including the changes in protein distribution among the nuclear and cytoplasmic compartments, recently discovered by an applicant) will be investigated;(b) The factors controlling implementation of the defensive responses to viral infection. In particular the steps that couple viral components with upstream apoptotic reactions (e. g., such as activation of initiating caspases) will be investigated. Contribution of cell-specific difference in the balance of proapoptotic and antiapoptotic factors to the character of the response to picornavirus infection will be studied;(c) The structure/function relationships in replicative (oriL, oriR, cre) and translational (IRES) cis-acting elements of picornavirus genomes. In particular, a new approach to elucidation of the structural stringency of the apparently conserved control elements will be exploited: either individual motifs or entire elements will be replaced, in the full-length viral genomes, by random sequences followed by selection and analysis of viable progeny. Also physical methods (e. g., NMR) will be used to solve the complex structure of the regulatory elements;(d) The mechanisms and biological significance of recombination between RNA molecules. Special attention will be paid to the mechanism of nonreplicative RNA recombination recently discovered by the applicants. The mechanisms of cleavage and ligation of the RNA fragments will be studied. Occurrence and significance of natural recombination between picornaviruses will be studied by both sequencing of genomes of circulating viruses and by analysis of sequence databases;(e) Development of new tools for RNA analysis in evolutionary and molecular epidemiological studies. In particular, an RT-PCR version of the molecular colony technique recently worked out by one of the applicant will be further developed and adapted for studying recombination between any viral and/or cellular RNAs. Possible advantages of cDNA microarray technique for fast and precise characterization of the genomes of circulating polioviruses will be evaluated.The advantage of the proposed cooperation consists in using diverse experimental objects to check the generality of the conclusions made; using diverse approaches to the solution of the same problem, and sharing expertise, resources and ideas. The collaborative investigations, which have already been carried out, proved to be successful, as judged by the importance of the results obtained and published in high ranked journals. As the experience shows, such a collaborative program adds significantly to the progress of the research of each participating group. Programma(i) IC-INTAS - International Association for the promotion of cooperation with scientists from the independent states of the former Soviet Union (INTAS), 1993- Argomento(i) 4 - Life Sciences OPEN - OPEN Call Invito a presentare proposte Data not available Meccanismo di finanziamento Data not available Coordinatore University Medical Center Nijmegen Contributo UE Nessun dato Indirizzo Geert Grooteplein zuid 24 6500 HB Nijmegen Paesi Bassi Mostra sulla mappa Costo totale Nessun dato Partecipanti (3) Classifica in ordine alfabetico Classifica per Contributo UE Espandi tutto Riduci tutto M.P.Chumakov Institute of Poliomyelitis & Viral Encephalitides Russia Contributo UE Nessun dato Indirizzo Kievskoye Shosse, 27 km 142782 Moscow Region Mostra sulla mappa Costo totale Nessun dato Russian Academy of Sciences Russia Contributo UE Nessun dato Indirizzo Institutskaya 4 142292 Pushchino, Moscow Region Mostra sulla mappa Costo totale Nessun dato UNIVERSITY OF HELSINKI Finlandia Contributo UE Nessun dato Indirizzo Haartmaninkatu 3 00014 HELSINKI Mostra sulla mappa Costo totale Nessun dato
