Caracterización de proteína-fosfatasas de tipo 1 de Leishmania infantum y estudio mediante RNA-Seq de las alteraciones en la expresión génica diferencial inducidas en líneas Knock-in

Abstract

Leishmania infantum (Kinetoplastida: Trypanosomatidae) is the causative agent of visceral leishmaniasis in the Mediterranean Basin, where dogs are the main reservoir. The infection is transmitted to the mammalian host by the bite of dipteran of the subfamily Phlebotominae. The life cycle of the parasite is digenetic and two stages alternate: the motile promastigote stage which develops in the invertebrate host and the non-motile amastigote stage, which multiplies within phagocytes of the mammalian host. Genome sequencing of the main Leishmania species was published between 2005 and 2007, and the development of high-throughput techniques, such as DNA microarrays, quantitative proteomics, and more recent methods based on next generation sequencing, have made exhaustive analysis of gene expression profiles of the parasite possible throughout the life cycle. This has been performed in order to find possible virulence factors involved in pathogenicity and/or possible therapeutic targets and vaccine candidates. In a previous comparative transcriptomics analysis performed in the laboratory of Molecular Parasitology of the Biological Research Center (CSIC), differential expression of the genes LinJ.34.0840 and LinJ.15.0240 encoding protein phosphatases 1 (PP1) was described in promastigotes isolated from the vector Phlebotomus perniciosus. Even though it has become clear that the phosphorylation status of protein is remarkable during the differentiation processes of the parasite, funcionality of PP1 proteins from Leishmania spp. is not known accurately so far, as well as the signalling pathways in general. For this reason, characterization of these proteins was proposed to develop this doctoral thesis, which has been carried out in the laboratory mentioned...