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Investigating the Role of Histone H4 Acetylation in RNA Splicing in Saccharomyces cerevisiae
Author(s) -
Kopew Jessica S,
Mendizabal Adriana G,
Lin Mengjia M,
Kress Tracy L
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.459.6
Subject(s) - rna splicing , biology , histone h4 , splicing factor , genetics , exonic splicing enhancer , histone h2a , histone , chromatin , chromatin remodeling , microbiology and biotechnology , gene , rna
The ability of RNA splicing and transcription to take place simultaneously allows for efficient coordination and regulation within cells. Few known connections exist between chromatin modifications, which regulate transcription, and RNA splicing in the yeast Sacchraomyces cerevisiae , as well as in metazoa1,2, leaving much to explore. Our previous research has shown that genetic interactions exist between splicing protein genes and the NuA4 complex genes in S. cerevisiae . In addition, we found that deletion or mutation of these NuA4 complex genes impact RNA splicing. Nua4 plays a key role in regulating transcription by acetylating histone H4; following this, the Swr1 chromatin remodeling complex is recruited to the acetylated histone and ultimately deposits H2A.Z in place of H2A, allowing for transcriptional activity3–5. Given these findings, our current research is focused on elucidating whether specific acetylation of histone H4 is required for coordinating splicing with transcription. We constructed an array of double mutant yeast strains comprising a deletion of an essential splicing factor gene and a point mutation in either H4K5, H4K8, H4K12, or H4K16 thus preventing a chemical modification. We conducted genetic interaction studies to identify synthetic sick/lethal, or suppressive genetic interactions between the two genes. The majority of the double mutant growth phenotypes reveal synthetic sick/lethal interactions when compared to single mutant strains, suggesting a functional link between H4 acetylation and RNA splicing. Using RT‐qPCR we determined that mutation of H4 resulted in a reduction in pre‐mRNA splicing. In addition, mutation of H4 exacerbates the splicing defects observed in splicing factor deletion strains, which is consistent with the genetic interactions. Our future research will focus on identifying the specific mechanism by which the histone H4 acetylation is functioning to regulate RNA splicing. To this end we are currently using chromatin immunoprecipitation to examine the impact of these H4 mutations on the recruitment of splicing factors. Support or Funding Information Research Corporation for the Advancement of Science (Cottrell College Science award no. 20186) National Institutes of Health (R15GM122026). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .