Role of the cellular decapping activator LSM1-7 complex in the replication of positive-strand RNA viruses

By using the ability of the positive-strand RNA ((+)RNA) virus BMV to replicate in yeast it was previously shown that subunits of the LSm1-7 ring, as well as Pat1 and Dhh1 play an essential role in the transit of the BMV genome from translation to replication. In non-infected cells, these proteins m...

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Detalles Bibliográficos
Autor: Galão, Rui Pedro Ribeiro
Tipo de recurso: tesis doctoral
Estado:Versión publicada
Fecha de publicación:2010
País:España
Institución:CBUC, CESCA
Repositorio:TDR. Tesis Doctorales en Red
OAI Identifier:oai:www.tdx.cat:10803/7222
Acceso en línea:http://www.tdx.cat/TDX-0207111-144624
http://hdl.handle.net/10803/7222
Access Level:acceso abierto
Palabra clave:ARN missatger
Virus de l'hepatitis C
Actina
Antiviral
Antibiòtic
Messenger RNA
MicroRNA
Hepatitis C virus
HCV
Actin
Brome Mosaic Virus
BMV
578
Descripción
Sumario:By using the ability of the positive-strand RNA ((+)RNA) virus BMV to replicate in yeast it was previously shown that subunits of the LSm1-7 ring, as well as Pat1 and Dhh1 play an essential role in the transit of the BMV genome from translation to replication. In non-infected cells, these proteins mediate the transition of cellular mRNAs from a translational to a non-translational state by activating decapping in the 5'-3' - deadenylation-dependent mRNA decay pathway. Given the conservation of this pathway from yeast to humans and the common need of all (+)RNA viruses to regulate the transition of their genomes from active translation to a translationally inactive state to allow replication, an exciting possibility, and our working hypothesis, was that LSm1-7, Dhh1 and Pat1 are used not only by BMV to replicate in yeast but also by human (+) RNA viruses, such as HCV, to replicate in mammalian cells. Furthermore, given the key role of these proteins in a common step to all (+)RNA viruses, it is essential to characterize the not yet defined molecular mechanisms associated with such function. In this regard, we also hypothesized that the LSm1-7 complex, as member of the Sm family of proteins, would directly interact with viral genomes of (+)RNA viruses in order to play their role in the virus life cycle in a similar way that other family counterparts directly interact with their RNA targets in order to achieve their different cellular functions. In this work we were able to confirm both hypothesis showing that human homologues of the upper mentioned proteins LSm1-7, Rck/p54 and PatL1, are required for HCV RNA translation and replication. Additionally, we also showed that reconstituted LSm1-7 complexes specifically recognize important signals, either in BMV or HCV genomes, that regulate their translation and/or replication. These observations constitute the first evidence that the LSm1-7 complex is able to directly interact with viral genomes representing also novel LSm1-7 interaction sites. Given the common replication strategies of (+)RNA viruses and the conserved cellular functions of LSm1-7, Pat1 and Dhh1 from yeast to humans, our findings pinpoint a weak spot that may be exploited to generate broad-spectrum antiviral drugs.