IN02, A Positive Regulator of Lipid Biosynthesis, Is Essential for the Formation of Inducible Membranes in Yeast

Block-Alper, Laura; Webster, Paul; Zhou, Xianghong Jasmine; Supeková, Lubica; Wong, Wing Hung; Schultz, Peter G.; Meyer, David I.

doi:10.1091/mbc.01-07-0366

Cited by 28 publications

(26 citation statements)

References 61 publications

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“…1-2). INO2 and INO4 are activators of phospholipid biosynthesis genes, and, thus, their deletions are known to reduce phospholipid levels and prevent membrane proliferation (Block-Alper et al, 2002; Schuck et al, 2009). The ino2Δ ino4Δ yeast is viable due to the base-level phospholipid biosynthesis that occurs in the absence of Ino2/Ino4 transcription activators.…”

Section: Discussionmentioning

confidence: 99%

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Inhibition of phospholipid biosynthesis decreases the activity of the tombusvirus replicase and alters the subcellular localization of replication proteins

2011

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Replication of plus-strand RNA viruses depends on lipids present in cellular membranes. Recent genome-wide screens have revealed that eight phospholipid biosynthesis genes affected the replication of Tomato bushy stunt virus (TBSV) in yeast model host. To test the importance of phospholipids in TBSV replication, we studied one of the identified genes, namely INO2, which forms a heterodimer with Ino4, and is a transcription activator involved in regulation of phospholipid biosynthesis. Deletion of INO2, or double deletion of INO2/INO4, reduced TBSV replication and inhibited the activity of the viral replicase complex. In addition, the stability of the viral replication protein is decreased as well as the localization pattern of the viral protein changed dramatically in ino2Δ ino4Δ yeast. Over-expression of Opi1, a repressor of Ino2 and phospholipid biosynthesis, also inhibited TBSV RNA accumulation. In contrast, over-expression of Ino2 stimulated TBSV RNA accumulation. We also observed an inhibitory effect on Flock house virus (FHV) replication and the reduced stability of the FHV replication protein in ino2Δ ino4Δ yeast. These data are consistent with the important role of phospholipids in RNA virus replication.

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Section: Discussionmentioning

confidence: 99%

“…Ino2 is a basic helix- loop-helix transcription activator for phospholipid synthesis genes (Block-Alper et al, 2002). The phospholipid biosynthesis in S. cerevisiae is based on biochemical pathways conserved in higher eukaryotes (Carman and Han, 2009; Daum et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Inhibition of phospholipid biosynthesis decreases the activity of the tombusvirus replicase and alters the subcellular localization of replication proteins

2011

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“…Cells stained with DiOC 6 were analyzed immediately. Staining was carried out as described by Block-Alper et al (25).…”

Section: Methodsmentioning

confidence: 99%

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

Murata¹,

Watanabe²,

Sato³

et al. 2003

Journal of Biological Chemistry

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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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“…If the cell density was A 600 > 1.0 at harvesting, the cell suspension was diluted with the TE buffer to an A 600 of ∼1.0. Cells stained with DiOC6 were analyzed immediately ( Block-Alper et al 2002). For Nile Red staining, yeast cells in stationary phase were washed and resuspended in phosphate-buffered saline (PBS).…”

Section: Microscopical Analysismentioning

confidence: 99%

Endoplasmic reticulum stress and calcium imbalance are involved in cadmium-induced lipid aberrancy in Saccharomyces cerevisiae

Rajakumar

Bhanupriya

Ravi

2016

Cell Stress and Chaperones

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The endoplasmic reticulum is the key organelle which controls protein folding, lipid biogenesis, and calcium (Ca 2+ ) homeostasis. Cd exposure in Saccharomyces cerevisiae activated the unfolded protein response and was confirmed by the increased Kar2p expression. Cd exposure in wild-type (WT) cells increased PC levels and the PC biosynthetic genes. Deletion of the two phospholipid methyltransferases CHO2 and OPI3 modulated PC, TAG levels and the lipid droplets with cadmium exposure. Interestingly, we noticed an increase in the calcium levels upon Cd exposure in the mutant cells. This study concluded that Cd interrupted calcium homeostasis-induced lipid dysregulation leading to ER stress.

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IN02, A Positive Regulator of Lipid Biosynthesis, Is Essential for the Formation of Inducible Membranes in Yeast

Cited by 28 publications

References 61 publications

Inhibition of phospholipid biosynthesis decreases the activity of the tombusvirus replicase and alters the subcellular localization of replication proteins

Inhibition of phospholipid biosynthesis decreases the activity of the tombusvirus replicase and alters the subcellular localization of replication proteins

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

Endoplasmic reticulum stress and calcium imbalance are involved in cadmium-induced lipid aberrancy in Saccharomyces cerevisiae

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