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Microbiol Biotechnol Rep | Volume 1, Issue 2
November 16-17, 2017 Atlanta, Georgia, USA
Annual Congress on
Mycology and Fungal Infections
High-throughput screening to identify regulators of meiosis-specific gene expression in
Saccharomyces cerevisiae
Yona Kassir
Technion-Israel Institute of Technology, Israel
T
ranscriptional regulation is a key mechanism that controls the fate and response of cells to diverse signals.
Therefore, the identification of the signal transduction pathways as well as the DNA-binding proteins, which
mediate these signals, is a crucial step in elucidating how cell fate is regulated, and how can we perturb it. In
the talk, I will discuss bioinformatics and functional high-throughput genomic approaches. Our model system is
the budding yeast
Saccharomyces cerevisiae
, and the
IME1
gene that encodes the master regulator of meiosis.
High throughput technology, based on fluorescent reporters (R-SGA), allows the screening of an array of all
viable yeast gene deletion mutants. This protocol promoted the identification of too many potential transcription
factors, and signal transduction factors. The main problem we faced was to discriminate between false and true
regulators. Bioinformatic analysis identified potential cis-regulatory sequences with perfect homology to
known transcription factors (TF). However, these consensuses and their corresponding TFs were found to
be nonfunctional in the R-SGA analysis. Moreover, many TFS were shown to bind to a non-perfect site.
The most rewarding approach was to examine a TF only if its known upstream regulators were also found in the
screen. Specific examples will be given. In conclusion, our results support the view that although bioinformatic
analysis can provide a useful guide, functional assays are required for accurate identification of TF-binding site
interactions in complex promoters.
Biography
Yona Kassir is currently working as a Professor Emeritus at Technion-Israel Institute of Technology, Israel.
ykassir@tx.technion.ac.il