Involvement of flocculin in negative potential-applied ITO electrode adhesion of yeast cells.

Sumihiro Koyama,Yuji Hatada,Chiaki Kato,Takashi Toyofuku,Yukari Ohta,Yuichi Nogi,Takehiko Nagahama,Taishi Tsubouchi,Masaaki Konishi,Akihiro Tame,Tetsuya Miwa,Yuriko Nagano,Fumiyoshi Abe,Keiko Usui,Katsuyuki Uematsu,Richard Calderone

doi:10.1093/femsyr/fov064

Abstract

The purpose of this study was to develop novel methods for attachment and cultivation of specifically positioned single yeast cells on a microelectrode surface with the application of a weak electrical potential. Saccharomyces cerevisiae diploid strains attached to an indium tin oxide/glass (ITO) electrode to which a negative potential between −0.2 and −0.4 V vs. Ag/AgCl was applied, while they did not adhere to a gallium-doped zinc oxide/glass electrode surface. The yeast cells attached to the negative potential-applied ITO electrodes showed normal cell proliferation. We found that the flocculin FLO10 gene-disrupted diploid BY4743 mutant strain (flo10Δ /flo10Δ) almost completely lost the ability to adhere to the negative potential-applied ITO electrode. Our results indicate that the mechanisms of diploid BY4743 S. cerevisiae adhesion involve interaction between the negative potential-applied ITO electrode and the Flo10 protein on the cell wall surface. A combination of micropatterning techniques of living single yeast cell on the ITO electrode and omics technologies holds potential of novel, highly parallelized, microchip-based single-cell analysis that will contribute to new screening concepts and applications.

Highlights

The first complete genome sequence of the yeast Saccharomyces cerevisiae was published (Goffeau et al 1996), enabling a wide variety of genetic engineering techniques and the strict functional analyses of proteins (Giaever et al 2002; Shibasaki, Maeda and Ueda 2009; Breker, Gymrek and Schuldiner 2013)
Saccharomyces cerevisiae diploid strains attached to an indium tin oxide/glass (ITO) electrode to which a negative potential between −0.2 and −0.4 V vs. Ag/AgCl was applied, while they did not adhere to a gallium-doped zinc oxide/glass electrode surface
Our results indicate that the mechanisms of diploid BY4743 S. cerevisiae adhesion involve interaction between the negative potential-applied ITO electrode and the Flo10 protein on the cell wall surface

Summary

Introduction

The first complete genome sequence of the yeast Saccharomyces cerevisiae was published (Goffeau et al 1996), enabling a wide variety of genetic engineering techniques and the strict functional analyses of proteins (Giaever et al 2002; Shibasaki, Maeda and Ueda 2009; Breker, Gymrek and Schuldiner 2013). A heterozygous diploid mutant collection of approximately 6000 strains of S. cerevisiae, in each of which one copy of a single gene is deleted, is commercially available With this collection, it is possible to evaluate the role of each gene product in the response of cells to a drug (Lum et al 2004; Parsons et al 2004, 2006; Roberge 2008). Drug-induced haploinsufficiency profiling (HIP)/homozygous profiling (HOP) assay was one of the first assays to take advantage of parallelized growth strategy (Smith et al 2010). The HIP/HOP assay has been applied to identify the protein targets in numerous drugs and successfully employed in industry (Smith et al 2010; Giaever and Nislow 2014)

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Conclusion