In our lab at IRB Barcelona Lluis Ribas de Pouplana discovered a new mitochondrial protein in insects and called it SLIMP (seryl-tRNA synthetase-like insect mitochondrial protein). The gene coding for this protein has been pointed out in 2009 at Illinois laboratories. Here, Ambrus et al. discovered the presence of SLIMP gene (CG31133) from a genetic screen while analyzing the effects of its depletion in a de2f1 mutant. They deleted de2f1 gene in Drosophila melanogaster and observed an arrest of the cell cycle accompanied by a reduction in viability, thus they screened which gene deletion could recover the phenotype. As a result, they produced double knock-outs of de2f1 plus several genes, among which there was CG31133. In our laboratory the transcriptomic of SLIMP-KD cells have been analyzed and several transcripts appeared to be regulated by SLIMP. The most likely explanation is that SLIMP is repressing an alternative pathway promoting cell cycle progression. Still and all, we do not know which of these modulated transcripts are direct targets of SLIMP and which are in common with dE2F pathway. The aim of this thesis is to validate that result producing three stable cell lines: SLIMP/dE2F1, SLIMP/dE2F2 and SLIMP/dE2F1/dE2F2 knock-downs. In that manner I will be able to compare the transcriptional analysis of these new cell lines with the one of SLIMP- KD and discriminate among the genes modulated by SLIMP and the ones modulated by both SLIMP and dE2F. The overall strategy starts with the production of two expression vectors for the transcription of two dsRNAs for dE2F1 and dE2F2, the transfection of S2 SLIMP-KD D. melanogaster embryo cells with the plasmids and the characterization of the phenotype.
Characterization of SLIMP, a protein related to the cell cycle
BONSEMBIANTE, AURELIANO
2021/2022
Abstract
In our lab at IRB Barcelona Lluis Ribas de Pouplana discovered a new mitochondrial protein in insects and called it SLIMP (seryl-tRNA synthetase-like insect mitochondrial protein). The gene coding for this protein has been pointed out in 2009 at Illinois laboratories. Here, Ambrus et al. discovered the presence of SLIMP gene (CG31133) from a genetic screen while analyzing the effects of its depletion in a de2f1 mutant. They deleted de2f1 gene in Drosophila melanogaster and observed an arrest of the cell cycle accompanied by a reduction in viability, thus they screened which gene deletion could recover the phenotype. As a result, they produced double knock-outs of de2f1 plus several genes, among which there was CG31133. In our laboratory the transcriptomic of SLIMP-KD cells have been analyzed and several transcripts appeared to be regulated by SLIMP. The most likely explanation is that SLIMP is repressing an alternative pathway promoting cell cycle progression. Still and all, we do not know which of these modulated transcripts are direct targets of SLIMP and which are in common with dE2F pathway. The aim of this thesis is to validate that result producing three stable cell lines: SLIMP/dE2F1, SLIMP/dE2F2 and SLIMP/dE2F1/dE2F2 knock-downs. In that manner I will be able to compare the transcriptional analysis of these new cell lines with the one of SLIMP- KD and discriminate among the genes modulated by SLIMP and the ones modulated by both SLIMP and dE2F. The overall strategy starts with the production of two expression vectors for the transcription of two dsRNAs for dE2F1 and dE2F2, the transfection of S2 SLIMP-KD D. melanogaster embryo cells with the plasmids and the characterization of the phenotype.The text of this website © Università degli studi di Padova. Full Text are published under a non-exclusive license. Metadata are under a CC0 License
https://hdl.handle.net/20.500.12608/42308