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Phosphorylation of SR Proteins and the Effect of Hydrogen Peroxide Stress on the Splicing Cycle in S. pombe Fission Yeast
Author(s) -
Davis Moriah Hope,
Piening Cole,
Marvin Micheal
Publication year - 2020
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.2020.34.s1.09641
Subject(s) - spliceosome , rna splicing , schizosaccharomyces pombe , sr protein , intron , splicing factor , yeast , schizosaccharomyces , biology , phosphorylation , microbiology and biotechnology , alternative splicing , exonic splicing enhancer , genetics , rna , exon , saccharomyces cerevisiae , gene
The spliceosome, a large and dynamic RNA‐protein complex, is responsible for effective pre‐mRNA splicing. Correct spliceosome assembly, function, and regulation have often been shown to involve SR proteins, which are dynamically phosphorylated. Alterations in the splicing process can lead to a variety of human diseases and the fission yeast, Schizosaccharomyces pombe, is one model organism for this research. It was previously determined that the conserved SR protein kinase in fission yeast, Dsk1 (Srpk1 in humans), is required for efficient splicing of introns. Previous identification of Dsk1 substrates and alanine mutation to prevent phosphorylation showed a minor splicing defect in the two fission yeast SR proteins (Srp1 and Srp2) under optimal growth conditions. Our current research takes a chemical‐genetic approach to exploring phosphorylation’s physical effects on specifically Srp1 and Srp2 protein substrates, and whether environmental stress, simulated by hydrogen peroxide exposure, can cause a higher retention rate of introns in a regulated way. Through the use of RT‐qPCR we can determine whether there is a splicing defect under the stress condition, and how phosphorylation of SR proteins affects the level of pre‐mRNA splicing defect. Support or Funding Information Previous work in Christine Guthrie’s lab in collaboration with Kevan Shokat and postdoctoral fellow Jesse Lipp, as well as previous work by Alex Piening of Rockhurst class 2018 and Clayton Carroll of Rockhurst class 2019. Thanks to Jeff Pleiss for microarray design as well as Nick Hertz & Alma Burlingame for mass spectrometry analysis. In addition, NIH NRSA postdoctoral fellowship for funding. Current funding and support provided by Rockhurst University.Rpl25a intron splicing in WT SR protein yeast strains and alanine mutated strains under optimal conditions.S. pombe cell responses due to increasing concentrations of hydrogen peroxide (Veal, E. 2014).