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| Author(s): |
Francisca dos Santos Borges
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Ribeirão Preto. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC) |
| Defense date: | 2022-03-04 |
| Examining board members: |
Angela Kaysel Cruz;
Fabiola Barbieri Holetz;
Giuseppe Palmisano
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| Advisor: | Angela Kaysel Cruz |
| Abstract | |
In the Leishmania parasite, most of the genes encoding ribosomal proteins (RPs) are present as two or more copies, and their transcripts have identical or similar coding regions (CDSs) and, in most cases, divergent untranslated regions (UTRs). The divergence in UTRs may be associated with a distinct regulation of expression for each CDS, and divergences within the CDSs, in turn, may result in functional differences between paralogs. RPs have been reported to exert extraribosomal functions (moonlight activities) in several organisms. Therefore, we decided to investigate ribosomal proteins in Leishmania by analyzing expression profiles of two RP genes duplicated in the genome of L. major. The genes coding for S16 ribosomal protein are duplicated as neighbor copies, RPS16_80 and RPS16_90; the transcripts have identical CDS and divergent UTRs. The copies of genes coding for the L13 ribosomal protein are located in different chromosomes, named RPL13_15 and RPL13_34, and their CDSs and UTRs are divergent. Herein, to study these proteins\' expression profiles, we generated transfectants with tagged RPs or knocked out RPs, using the CRISPR/Cas9 genomic editing system. Our data revealed that: 1) the expression of the two isoforms of RPS16 and RPL13 varies throughout development, with one of the isoforms being more expressed than the other; 2) there is a mechanism of paralog compensation; in the absence of one isoform of RPS16 and RPL13, the other has its expression increased, and 3) cell viability of all knockout parasites (for each one of the four genes) is increased under nutritional stress when compared to the control strain. We essayed to identify post-translational modifications in one of the RPL13 paralogs unsuccessfully. In conclusion, our results created essential tools to understand better the functional meaning of two ribosomal proteins\' coding genes duplication in Leishmania. (AU) | |
| FAPESP's process: | 19/05257-6 - Outcomes of the duplication of ribosomal protein coding genes in Leishmania |
| Grantee: | Francisca dos Santos Borges |
| Support Opportunities: | Scholarships in Brazil - Master |