Systematic profiling of subtelomeric silencing factors in budding yeast
Por:
Juárez-Reyes A., Avelar-Rivas J.A., Hernandez-Valdes J.A., Hua B., Campos S.E., González J., González A., Springer M., Mancera E., DeLuna A.
Publicada:
1 ene 2023
Resumen:
Subtelomeric gene silencing is the negative transcriptional regulation of genes located close to telomeres. This phenomenon occurs in a variety of eukaryotes with salient physiological implications, such as cell adherence, virulence, immune-system escape, and ageing. The process has been widely studied in the budding yeast Saccharomyces cerevisiae, where genes involved in this process have been identified mostly on a gene-by-gene basis. Here, we introduce a quantitative approach to study gene silencing, that couples the classical URA3 reporter with GFP monitoring, amenable to high-throughput flow cytometry analysis. This dual silencing reporter was integrated into several subtelomeric loci in the genome, where it showed a gradual range of silencing effects. By crossing strains with this dual reporter at the COS12 and YFR057W subtelomeric query loci with gene-deletion mutants, we carried out a large-scale forward screen for potential silencing factors. The approach was replicable and allowed accurate detection of expression changes. Results of our comprehensive screen suggest that the main players influencing subtelomeric silencing were previously known, but additional potential factors underlying chromatin conformation are involved. We validate and report the novel silencing factor LGE1, a protein with unknown molecular function required for histone H2B ubiquitination. Our strategy can be readily combined with other reporters and gene perturbation collections, making it a versatile tool to study gene silencing at a genome-wide scale. © The Author(s) 2023. Published by Oxford University Press on behalf of The Genetics Society of America.
Filiaciones:
Unidad de Genómica Avanzada (Langebio), Centro de Investigación y de Estudios Avanzados, 36824 Irapuato, Guanajuato, Mexico
Departamento de Ingeniería Genética, Centro de Investigación y de Estudios Avanzados, Unidad Irapuato, 36824 Irapuato, Guanajuato, Mexico
Nouryon Chemicals Research Centre, Expert Capability Center Deventer, 7418AJ Deventer, Netherlands
Department of Systems Biology, Harvard Medical School, Boston, MA 02115, United States
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, 04510 Ciudad de México, Mexico
Departamento de Biología Celular, Universidad Nacional Autónoma de México, Facultad de Ciencias, 04510 Ciudad de México, Mexico
Green Published, gold, All Open Access; Gold
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