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Metabolic state and cell cycle as determinants of facilitated uptake of genetic information by yeast Saccharomyces cerevisiae
Authors:Larisa Chaustova  Valė Miliukienė  Aurelijus Zimkus  Valdemaras Razumas
Affiliation:(1) Department of Bioelectrochemistry and Biospectroscopy, Institute of Biochemistry, LT-08662 Vilnius, Lithuania;(2) Department of Xenobiotics, Institute of Biochemistry, LT-08662 Vilnius, Lithuania;(3) Department of Biochemistry and Biophysics, Vilnius University, LT-01513 Vilnius, Lithuania
Abstract:The transformation efficiency of yeast cells during exponential growth might be characterised as undulatory. The aim of the study was to investigate the reason for the fluctuation in transformation efficiency of yeast Saccharomyces cerevisiae p63-DC5 cells during exponential growth. The heightened response to exogenous DNA was observed with the growing yeast culture when budded cells were predominant. To confirm this phenomenon we carried out synchronization of yeast cells with 10 mM hydroxyurea. Results showed that synchronous yeast cells in the S-phase of cell cycle have enhanced transformation efficiency. Furthermore, S. cerevisiae p63-DC5 cells in the S-phase were successfully transformed with plasmid pl13 in the absence of lithium acetate. We indicated that the permeability of yeast cells in the S-phase to tetraphenylphosphonium (TPP) cations was significantly higher than in asynchronous culture. The results of our study showed that the fluctuation in transformation efficiency was strictly dependent on the metabolic state of yeast cells and the capacity of the yeast cells to become competent was related to the S-phase of cell cycle.
Keywords:Saccharomyces cerevisiae  Transformation  Cell cycle  S-phase  Permeability  Tetraphenylphosphonium (TPP)
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