Our research outlined the physiological effects of ribosome biogenesis deficiencies. We studied null mutants of ribosomal proteins from both ribosomal subunits to: 1) Evaluate their growth and translational capacity, 2) Investigate impacts on protein translation, and 3) Explore potential links to the cytosolic ribosome quality control system
Through rigorous validation, we have established that these proteins are dispensable for cellular viability and that they perform protein synthesis. I have stablished Ribosome Profiling methodology within the host laboratory for the model organisms, Saccharomyces cerevisiae. The application of Ribosome Profiling as well as RNA-Seq methodologies, aimed to elucidate comprehensive insights into the translatome and transcriptome in different yeast strains. All the information generated will be uploaded to the public genomics data webpage Gene Expression Omnibus (GEO), a database repository of high throughput analysis that allows the use of unique identifiers. Among the various unresolved questions, we attempt to elucidate whether aberrant ribosomes exhibit a preference for translating specific pools of transcripts.
In summary of the advancement of the outlined objectives within the action plan and concurrent project commitments, the primary outcomes are summarized as follows: First, physiological characterization of ribosomal mutants from both small and large subunit, namely RiboEscapers, revealed their non-essential role in cell viability.
Second, standardizing growth conditions at 30°C proved conducive for all RiboEscapers.
Third, analysis of ribosomal subunit conditions, polysome assembly, and cellular translation through polysome profiling demonstrated the translation ability of RiboEscapers strains.
Fourth, successful implementation of the Ribosome profiling technique in the host laboratory for Saccharomyces cerevisiae encompassed optimizing cell growth, lysis, nuclease digestion, ribosome isolation, RNA extraction, and footprint generation.
The identity of the ribosomal protein mutants are listed in the fellowship technical report and will be available upon publication.
Following the completion of experimental procedures, we are presently in the stage awaiting the informatics analysis on the deep-sequenced datasets.
The upcoming publications resulting from our ongoing projects will integrate these datasets. Moreover, I am currently engaged in the revision of an article resulting from collaborative actions with Heidelberg University. Additionally, we are preparing a revision within the host laboratory.
During the project, I had the opportunity to present the RiboEscapers project through oral presentations at two national conferences: the IX Spanish Network Meeting on RNA (RIBORED 2022) and the workshop "Elements Empowering RNA Function: From Bench to Bedside".
The project offered extensive training, including proficiency in using Saccharomyces cerevisiae and genetic methods, financial management skills, security training, improved communication through presentations and lectures, mentoring and leadership development by supervising students, strengthened teaching abilities through lectures, and contributing to a successful national grant application
Finally I coordinated the Pint of Science festival in Seville for science outreach. I participated in IBISibiliza's open-door events by the Institute of Biomedicine. As a Marie Curie Ambassador, I gave a presentation on scientific careers at the WOW Science lecture series and another on Rare Diseases for the European Researchers' Night.
