Dynamics of Transcription Factors in live Saccharomyces cerevisiae investigated by Single Molecule Tracking Microscopy

Authors

• E Haque
• J Oladosu
• H Li
• D Zeldin
• SJ London

Abstract

Single Molecule Tracking (SMT) allows for the biophysical characterization of protein dynamics in in vivo by providing direct information on the behavior of individual molecules. Saccharomyces cerevisiae or budding yeast, a unicellular eukaryote with a small and completely sequenced genome and the awesome powers of yeast genetics, is an attractive model organism for SMT. We used a custom-built Highly Inclined Laminated Optical (HILO) sheet illumination for SMT analysis of yeast transcription factors Ace1p and HSF1, RNA polymerase II, and histone H3 in live cells. For quantification, we applied to single molecule tracks pEM analysis which is an algorithm that determines the distinct mobility states of the molecules. We compared parameters of biophysical dynamics for each protein. Preliminary data reveal distinct mobility states for yeast Histone H3 protein compared to RNA Pol II, Transcription Factor Ace1p, and Heat Shock Factor 1 proteins in the presence and absence of heat shock conditions. The results presented here provides a foundation for characterizing dynamics of other factors involved in transcription initiation and elongation.

Scientific Focus Area: Cell Biology

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