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Investigation of mutants with aberrant transcriptional regulation of OLE1
Author(s) -
Cribbs Lauren,
Filippelli Sara,
McDonough Virginia
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.s1.00607
Subject(s) - mutant , transcription (linguistics) , biology , transcriptional regulation , gene , gene expression , wild type , microbiology and biotechnology , genetics , philosophy , linguistics
The OLE1 gene in Saccharomyces cerevisiae encodes the Δ9 desaturase. Previously, we isolated mutants that affect the expression of OLE1 . In this work, we describe the phenotypes of two of these mutants, arv1 and nro (“ n o r egulation of OLE1” ). OLE1 promoter ‐URA3 ORF reporter gene assays have shown that both mutants express OLE1 in the presence of the normally repressive unsaturated fatty acid 18:2Δ9,12. Analysis of OLE1 RNA amounts by qPCR, indicate that the nro mutant has similar amounts of mRNA present to the wildtype control, however, the level of OLE1 mRNA is significantly lower in arv1 , and confirmed that it is not regulating expression in response to 18:2 feeding. OLE1 promoter deletion analysis resulted in decreased OLE1 transcription in arv1 compared to the wildtype, and did not regulate in response to 18:2 feeding. Surprisingly, OLE1 expression regulation by 18:2 is somewhat recovered with a OLE1 promoter deletion between −488 and −471 in arv1 cells, although this regulation is lost at −255 (same as wildtype). This assay also indicated that the OLE1 promoter region between the −225 and −142 may be important for nro regulation of OLE1 transcription. These results indicate that the arv1 and nro mutants do have an effect on the transcriptional regulation of the OLE1 gene. Support or Funding Information Research reported was supported by the National Institute Of General Medical Sciences of the National Institutes of Health under Award Number R15GM132853