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Investigating the GADPH‐Sir2 interaction in S. cerevisiae (763.2)
Author(s) -
Palmer Claire
Publication year - 2014
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.28.1_supplement.763.2
Subject(s) - acetylation , gene silencing , chromatin immunoprecipitation , glyceraldehyde 3 phosphate dehydrogenase , saccharomyces cerevisiae , histone deacetylase , hdac8 , histone , mutant , immunoprecipitation , lysine , biology , trichostatin a , sirt3 , microbiology and biotechnology , yeast , biochemistry , dehydrogenase , gene , sirtuin , enzyme , gene expression , amino acid , promoter
The yeast glyceraldehyde‐phosphate dehydrogenase (GAPDH), Tdh3, has been found to interact with the silencing protein Sir2 in S. cerevisiae. Sir2 is a protein deacetylase that removes acetyl groups from the tails of histones H3 and H4. Deacetylated histones are associated with more densely packed chromatin and thus, transcriptional silencing. The Sir2‐GAPDH interaction is being investigated in order to further understand this connection between metabolism and gene silencing. We are interested in determining what part of Sir2 is necessary for this interaction with Tdh3. To this end, Sir2 mutant strains for use in coimmunoprecipitation experiments have been created. We hope to identify strains in which coimmunoprecipitation of Sir2 with Tdh3 is unsuccessful, indicating that these strains are missing a part of Sir2 that is essential for Tdh3 interaction. We are also investigating the possibility that Sir2 could be interacting with Tdh3 in its capacity as a deacetylase. A previous genetic screen suggests that Sir2 may also interact with two ribosomal proteins, Rpl22a and Rps7a. Tdh3, a similar GAPDH protein, Tdh2, Rpl22a and Rps7a are therefore being tested for acetylation by immunoprecipitatation and probing with an anti‐acetyl lysine antibody. Thus far, no evidence of acetylation on Tdh3 has been observed. However, further experiments are necessary to confirm this result. Finally, to better understand Tdh3’s role in silencing, we are creating a library of Tdh3 mutants defective for promoting silencing. Grant Funding Source : Supported by National Science Foundation