Dimethyl Sulfoxide Exposure Facilitates Phospholipid Biosynthesis and Cellular Membrane Proliferation in Yeast Cells

Abstract

Me 2 SO is a polar solvent that is widely used in biochemistry, pharmacology, and industry. Although there are several reports in the literature concerning the biological effects of Me 2 SO, the total cellular response remains unclear. In this paper, DNA microarray technology combined with the hierarchical clustering bioinformatics tool was used to assess the effects of Me 2 SO on yeast cells. We found that yeast exposed to Me 2 SO increased phospholipid biosynthesis through up-regulated gene expression. It was confirmed by Northern blotting that the level of INO1 and OPI3 gene transcripts, encoding key enzymes in phospholipid biosynthesis, were significantly elevated following treatment with Me 2 SO. Furthermore, the phospholipid content of the cells increased during exposure to Me 2 SO as shown by conspicuous incorporation of a lipophilic fluorescent dye (3,3 -dihexyloxacarbocyanine iodide) into the cell membranes. From these results we propose that Me 2 SO treatment induces membrane proliferation in yeast cells to alleviate the adverse affects of this chemical on membrane integrity.Dimethyl sulfoxide (Me 2 SO) 1 is widely used as a solvent in the chemical industry and as a cryoprotectant in biotechnology. It is present in the environment as a waste product of the paper industry and from the production of dimethyl sulfide (DMS) and also arises from the degradation of sulfur-containing pesticides (1). In addition, Me 2 SO forms naturally from photooxidation of DMS in the atmosphere and from degradation of DMS by phytoplankton in the marine environment (2). Because Me 2 SO has low volatility and is highly hygroscopic, it is rapidly scavenged from the atmosphere by rain and returned to earth, and thereby plays a role in the global sulfur cycle (1, 3).Microorganisms, including both prokaryotes and eukaryotes, have the capability of reducing Me 2 SO and can utilize it as a terminal electron acceptor during anaerobic growth (4 -6). Me 2 SO reductase from Escherichia coli and Rhodobacter sphaeroides catalyzes the reduction of Me 2 SO to DMS (7-9). This enzyme, which contains a molybdenum cofactor at the active center, is well characterized at the biochemical, biophysical, and molecular level and provides an excellent model system for investigating the structure and mechanism of electron transfer chain complexes (1, 10). Saccharomyces cerevisiae has a number of NADPH-dependent enzymes that, in conjunction with methionine sulfoxide reductase (MXR1), can reduce Me 2 SO to DMS (11).In E. coli, the combination of divalent cations as Ca 2ϩ and Me 2 SO has been shown to stimulate the efficiency of DNA transfer into the cell (12). In the case of mammalian cells, it is reported that the transfection efficiency is increased by Me 2 SO treatment after electroporation (13). The exact mechanism of how Me 2 SO increases membrane permeability leading to DNA uptake is unclear.In mammalian cells Me 2 SO (2% (v/v)) can induce morphological changes, for example in mouse erythroleukemic cells (14), or cause differentiation, for example ...

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