Proteomics

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Transcriptome and proteome changes triggered by overexpression of the transcriptional regulator Maf1 in the human pathogen Leishmania major


ABSTRACT: Maf1, originally described as a repressor of RNA polymerase III (RNAP III) transcription in yeast, participates in multiple functions across eukaryotes. However, the knowledge about Maf1in protozoan parasites is scarce. To initiate the study of Maf1 in Leishmania major, we generated a cell line that overexpresses this protein. Overexpression of Maf1 led to a significant reduction in the abundance of tRNAs, 5S rRNA and U4 snRNA, demonstrating that Maf1 regulates RNAP III activity in L. major. To further explore the roles played by Maf1 in this microorganism, global transcriptomic and proteomic changes due to Maf1 overexpression were determined using RNA-sequencing and label-free quantitative mass spectrometry. Compared to wild-type cells, differential expression was observed for 1082 transcripts (615 down-regulated and 467 up-regulated) and 205 proteins (132 down-regulated and 73 up-regulated) in the overexpressing cells. A correlation of 42% was found between transcriptomic and proteomic results. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that the differentially expressed genes and proteins are mainly involved in transcription, cell cycle regulation, lipid metabolism and transport, ribosomal biogenesis, carbohydrate metabolism, and cytoskeleton modification. Thus, our results suggest the involvement of Maf1 in the regulation of all these processes in L. major, as reported in other species, indicating that the functions performed by Maf1 were established early in eukaryotic evolution. Notably, our data also suggest the participation of L. major Maf1 in mRNA post-transcriptional control, a role that, to the best of our knowledge, has not been described in other organisms.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Leishmania Major Strain Friedlin

TISSUE(S): Cell Culture

SUBMITTER: Ebbing de Jong  

LAB HEAD: Santiago Martinez-Calvillo

PROVIDER: PXD050692 | Pride | 2024-08-27

REPOSITORIES: Pride

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20220711_Santiago_11.mgf Mgf
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20220711_Santiago_12.mgf Mgf
20220711_Santiago_12.raw Raw
20220711_Santiago_13.mgf Mgf
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Transcriptome and proteome changes triggered by overexpression of the transcriptional regulator Maf1 in the human pathogen Leishmania major.

Rivera-Rivas Luis A LA   Florencio-Martínez Luis E LE   Romero-Meza Gabriela G   Ortega-Ortiz Roberto C RC   Manning-Cela Rebeca G RG   Carrero Julio C JC   Nepomuceno-Mejía Tomás T   Martínez-Calvillo Santiago S  

FASEB journal : official publication of the Federation of American Societies for Experimental Biology 20240801 16


Maf1, originally described as a repressor of RNA polymerase III (RNAP III) transcription in yeast, participates in multiple functions across eukaryotes. However, the knowledge about Maf1 in protozoan parasites is scarce. To initiate the study of Maf1 in Leishmania major, we generated a cell line that overexpresses this protein. Overexpression of Maf1 led to a significant reduction in the abundance of tRNAs, 5S rRNA, and U4 snRNA, demonstrating that Maf1 regulates RNAP III activity in L. major. T  ...[more]

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