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Determining the Role, Expression and Interactions of FAP‐1 in S. cerevisiae , Cultivated in a Nitrogen‐limited Media
Author(s) -
Velez Astrid Carolina Rodriguez,
Matos Elsie Pares
Publication year - 2018
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.2018.32.1_supplement.666.7
Subject(s) - saccharomyces cerevisiae , nutrient , biology , cytoplasm , yeast , ammonium sulfate , nitrogen , gene expression , incubation , ammonium , gene , microbiology and biotechnology , biochemistry , chemistry , ecology , organic chemistry , chromatography
The protein FAP‐1 was found in the cytoplasm of Saccharomyces cerevisiae, and it confers resistance to the immunosuppressive drug and antifungal Rapamycin, by competing with the drug for the binding site, which is mutually exclusive for both, thus preventing the formation of the Fpr1p‐Rapamycin complex. This complex inhibits the products of the TORC1 which regulates essential pathways that control some cytoplasmic and nuclear events, and some nutrient regulated genes. In preliminary studies, it has been observed that FAP1 needs to be overexpressed in other to confer this resistance to the yeast. The complex FAP1:Fpr1p has been also observed in the absent of Rapamycin, especially when some nutrient levels are low. By using these observations, we proposed that the concentration of FAP1 might be inversely proportional to the availability of nutrients in culturing media. To prove this hypothesis, the strain CFY7‐PJ69‐6A of Saccharomyces cerevisiae will be cultured in Nitrogen‐limited media and at different time periods. The technique qPCR was used to quantify the amount of fap1 RNA available at different concentrations of ammonium sulfate (the nitrogen source), and at incubation points of 3 hrs, 6 hrs, 9 hrs and 12 hrs. We found that the expression of fap1 is inversely proportional to the availability of nitrogen in culturing media, whereas the growth and division was directly proportional to the nitrogen availability. The expression levels observed with the qPCR could be useful to optimize bioprocesses at industry by monitoring FAP1 in order to determine the availability of target nutrients in media in yeast cultures. Currently, we are interested in the identification of protein‐DNA interactions via Chromatin Immunoprecipitation, to determine which genes this transcription factor might regulate. Support or Funding Information This work is supported by the NIH‐MARC‐U‐STAR Grants: 5T34GM008419‐25 and 2T34GM008419‐26, and Puerto Rico's Louis Stoke Alliance for Minority Participation. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .