scholarly journals Crz1p Regulates pH Homeostasis in Candida glabrata by Altering Membrane Lipid Composition

2016 ◽  
Vol 82 (23) ◽  
pp. 6920-6929 ◽  
Author(s):  
Dongni Yan ◽  
Xiaobao Lin ◽  
Yanli Qi ◽  
Hui Liu ◽  
Xiulai Chen ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Lipid Composition ◽  
Candida Glabrata ◽  
Membrane Integrity ◽  
Membrane Lipid ◽  
Low Ph ◽  
Wild Type ◽  
Content Type ◽  
Membrane Lipid Composition ◽  
Acidic Conditions

ABSTRACTThe asexual facultative aerobic haploid yeastCandida glabratais widely used in the industrial production of various organic acids. To elucidate the physiological function of theC. glabratatranscription factor Crz1p (CgCrz1p) and its role in tolerance to acid stress, we deleted or overexpressed the corresponding gene,CgCRZ1. Deletion ofCgCRZ1resulted in a 60% decrease in the dry weight of cells (DCW) and a 50% drop in cell viability compared with those of the wild type at pH 2.0. Expression of lipid metabolism-associated genes was also significantly downregulated. Consequently, the proportion of C18:1fatty acids, the ratio of unsaturated to saturated fatty acids, and the ergosterol content decreased by 30%, 46%, and 30%, respectively. Additionally, membrane integrity, fluidity, and H+-ATPase activity were reduced by 45%, 9%, and 50%, respectively. In contrast, overexpression of CgCrz1p increased C18:1and ergosterol contents by 16% and 40%, respectively. Overexpression also enhanced membrane integrity, fluidity, and H+-ATPase activity by 31%, 6%, and 20%, respectively. Moreover, in the absence of pH buffering, the DCW and pyruvate titers increased by 48% and 60%, respectively, compared to that of the wild type. Together, these results suggest that CgCrz1p regulates tolerance to acidic conditions by altering membrane lipid composition inC. glabrata.IMPORTANCEThis study provides insight into the metabolism ofCandida glabrataunder acidic conditions, such as those encountered during the industrial production of organic acids. We found that overexpression of the transcription factor CgCrz1p improved viability, biomass, and pyruvate yields at a low pH. Analysis of plasma membrane lipid composition indicated that CgCrz1p might play an important role in its integrity and fluidity and that it enhanced the pumping of protons in acidic environments. We propose that altering the structure of the cell membrane may provide a successful strategy for increasingC. glabrataproductivity at a low pH.

scholarly journals Med15B Regulates Acid Stress Response and Tolerance in Candida glabrata by Altering Membrane Lipid Composition

2017 ◽  
Vol 83 (18) ◽  
Author(s):  
Yanli Qi ◽  
Hui Liu ◽  
Jiayin Yu ◽  
Xiulai Chen ◽  
Liming Liu
Keyword(s):  
Fatty Acids ◽  
Atpase Activity ◽  
Lipid Composition ◽  
Candida Glabrata ◽  
Membrane Integrity ◽  
Acid Stress ◽  
Membrane Lipid ◽  
Low Ph ◽  
Content Type ◽  
Membrane Lipid Composition

ABSTRACT Candida glabrata is a promising producer of organic acids. To elucidate the physiological function of the Mediator tail subunit Med15B in the response to low-pH stress, we constructed a deletion strain, C. glabrata med15BΔ, and an overexpression strain, C. glabrata HTUΔ/CgMED15B. Deletion of MED15B caused biomass production, glucose consumption rate, and cell viability to decrease by 28.3%, 31.7%, and 26.5%, respectively, compared with those of the parent (HTUΔ) strain at pH 2.0. Expression of lipid metabolism-related genes was significantly downregulated in the med15BΔ strain, whereas key genes of ergosterol biosynthesis showed abnormal upregulation. This caused the proportion of C18:1 fatty acids, the ratio of unsaturated to saturated fatty acids (UFA/SFA), and the total phospholipid content to decrease by 11.6%, 27.4%, and 37.6%, respectively. Cells failed to synthesize fecosterol and ergosterol, leading to the accumulation and a 60.3-fold increase in the concentration of zymosterol. Additionally, cells showed reductions of 69.2%, 11.6%, and 21.8% in membrane integrity, fluidity, and H+-ATPase activity, respectively. In contrast, overexpression of Med15B increased the C18:1 levels, total phospholipids, ergosterol content, and UFA/SFA by 18.6%, 143.5%, 94.5%, and 18.7%, respectively. Membrane integrity, fluidity, and H+-ATPase activity also increased by 30.2%, 6.9%, and 51.8%, respectively. Furthermore, in the absence of pH buffering, dry weight of cells and pyruvate concentrations were 29.3% and 61.2% higher, respectively, than those of the parent strain. These results indicated that in C. glabrata, Med15B regulates tolerance toward low pH via transcriptional regulation of acid stress response genes and alteration in lipid composition. IMPORTANCE This study explored the role of the Mediator tail subunit Med15B in the metabolism of Candida glabrata under acidic conditions. Overexpression of MED15B enhanced yeast tolerance to low pH and improved biomass production, cell viability, and pyruvate yield. Membrane lipid composition data indicated that Med15B might play a critical role in membrane integrity, fluidity, and H+-ATPase activity homeostasis at low pH. Thus, controlling membrane composition may serve to increase C. glabrata productivity at low pH.

scholarly journals Candida glabrata Yap6 Recruits Med2 To Alter Glycerophospholipid Composition and Develop Acid pH Stress Resistance

2020 ◽  
Vol 86 (24) ◽  
Author(s):  
Pei Zhou ◽  
Xiaoke Yuan ◽  
Hui Liu ◽  
Yanli Qi ◽  
Xiulai Chen ◽  
...  
Keyword(s):  
Cell Growth ◽  
Type Strain ◽  
Candida Glabrata ◽  
Wild Type Strain ◽  
Membrane Integrity ◽  
Low Ph ◽  
Wild Type ◽  
Content Type ◽  
Acid Ph ◽  
Ph Stress

ABSTRACT Candida glabrata is a high-performance microbial cell factory for the production of organic acids. To elucidate the role of the C. glabrata Mediator tail subunit Med2 (CgMed2) at pH 2.0, we deleted or overexpressed CgMed2 and used transcriptome analysis to identify genes that are regulated by CgMed2. At pH 2.0, the deletion of CgMed2 resulted in a cell growth decrease of 26.1% and a survival decrease of 32.3%. Overexpression of CgMed2 increased cell growth by 12.4% and cell survival by 5.9% compared to the wild-type strain. Transcriptome and phenotypic analyses identified CgYap6 as a transcription factor involved in acid pH stress tolerance. Deletion of CgYap6 caused growth defects, whereas its overexpression enhanced cell growth at pH 2.0. Furthermore, total glycerophospholipid content and membrane integrity decreased by 33.4% and 21.8%, respectively, in the CgMed2Δ strain; however, overexpression of CgMed2 increased the total glycerophospholipid content and membrane integrity by 24.7% and 12.1%, respectively, compared with those of the wild-type strain at pH 2.0. These results demonstrated that under acid pH stress, CgMed2 physically interacts with CgYap6, which translocates from the cytoplasm to the nucleus after being phosphorylated by the protein kinase CgYak1. Once in the nucleus, CgYap6 recruits CgMed2 to express glycerophospholipid-related genes. Our study elucidated the function of CgMed2 under acid pH stress and provides a potential strategy to equip Candida glabrata with low-pH resistance during organic acid fermentation. IMPORTANCE This study investigated the function of the Mediator tail subunit CgMed2 in C. glabrata under low-pH stress. The protein kinase CgYak1 activates CgYap6 for the recruitment of CgMed2, which in turn increases glycerophospholipid content and membrane integrity to confer low-pH stress tolerance. This study establishes a new link between the Mediator tail subunit and transcription factors. Overall, these findings indicate that CgMed2 is a novel target to induce the low-pH stress response in C. glabrata.

scholarly journals CgHog1-Mediated CgRds2 Phosphorylation Alters Glycerophospholipid Composition To Coordinate Osmotic Stress in Candida glabrata

2019 ◽  
Vol 85 (6) ◽  
Author(s):  
Chengjin Wu ◽  
Jiali Zhang ◽  
Guoxing Zhu ◽  
Rui Yao ◽  
Xiulai Chen ◽  
...  
Keyword(s):  
Cell Growth ◽  
Osmotic Stress ◽  
Cell Survival ◽  
Type Strain ◽  
Candida Glabrata ◽  
Wild Type Strain ◽  
Membrane Integrity ◽  
Wild Type ◽  
Content Type ◽  
Zinc Cluster

ABSTRACT Under stress conditions, Hog1 is required for cell survival through transiently phosphorylating downstream targets and reprogramming gene expression. Here, we report that Candida glabrata Hog1 (CgHog1) interacts with and phosphorylates CgRds2, a zinc cluster transcription factor, in response to osmotic stress. Additionally, we found that deletion of CgRDS2 led to decreases in cell growth and cell survival by 23.4% and 39.6%, respectively, at 1.5 M NaCl, compared with levels of the wild-type strain. This is attributed to significant downregulation of the expression levels of glycerophospholipid metabolism genes. As a result, the content of total glycerophospholipid decreased by 30.3%. Membrane integrity also decreased 47.6% in the Cgrds2Δ strain at 1.5 M NaCl. In contrast, overexpression of CgRDS2 increased the cell growth and cell survival by 10.2% and 6.3%, respectively, owing to a significant increase in the total glycerophospholipid content and increased membrane integrity by 27.2% and 12.1%, respectively, at 1.5 M NaCl, compared with levels for the wild-type strain. However, a strain in which the CgRDS2 gene encodes the replacement of Ser64 and Thr97 residues with alanines (Cgrds22A), harboring a CgRds2 protein that was not phosphorylated by CgHog1, failed to promote glycerophospholipid metabolism and membrane integrity at 1.5 M NaCl. Thus, the above results demonstrate that CgHog1-mediated CgRds2 phosphorylation enhanced glycerophospholipid composition and membrane integrity to resist osmotic stress in C. glabrata. IMPORTANCE This study explored the role of CgHog1-mediated CgRds2 phosphorylation in response to osmotic stress in Candida glabrata. CgHog1 interacts with and phosphorylates CgRds2, a zinc cluster transcription factor, under osmotic stress. Phosphorylated CgRds2 plays an important role in increasing glycerophospholipid composition and membrane integrity, thereby enhancing cell growth and survival.

Effects of temperature on the structure and metabolism of cell membranes in fish

1984 ◽  
Vol 246 (4) ◽  
pp. R460-R470 ◽  
Author(s):  
J. R. Hazel
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Cold Exposure ◽  
Lipid Composition ◽  
Membrane Lipid ◽  
Effect Of Temperature ◽  
Salmo Gairdneri ◽  
Homeoviscous Adaptation ◽  
Cold Temperatures ◽  
Membrane Lipid Composition

The metabolic adjustments responsible for the “homeoviscous adaptation” of membrane lipid composition in fish are examined with special reference to the rainbow trout, Salmo gairdneri. The percentage of fatty acid lipogenesis attributable to unsaturates was elevated after an acute drop in temperature but declined with continued cold exposure (i.e., cold acclimation). In contrast, selected desaturation reactions [particularly those involved in the production of polyunsaturated fatty acids (PUFA) of the n-3 and/or n-6 families] proceeded more rapidly in cold-than in warm-acclimated trout. Different time courses for the change in monoene and PUFA levels of hepatic microsomal membranes during thermal acclimation suggest that the various desaturase enzymes contribute to the acclimatory response at different times. Certain fatty acids, particularly the delta 5-desaturation products of the n-3 (20:5 delta 5,8,11,14,17) and n-6 (20:4 delta 5,8,11,14) series, were preferentially incorporated into phospholipids at cold temperatures and by cold-acclimated trout, due in part to the direct effect of temperature on the substrate preferences of the phospho- and acyltransferase enzymes of de novo phospholipid biosynthesis; however, chain length rather than degree of unsaturation per se may determine the temperature-dependent pattern of fatty acid incorporation. Both acute and chronic cold exposure elevated the incorporation of PUFA into phosphatidylserine (PS), suggesting that the conversion of PS to phosphatidylethanolamine (PE) may be activated at cold temperatures. The rate of homeoviscous adaptation appears to be limited by the rate of membrane lipid turnover, which although generally positively correlated with acclimation temperature, did vary depending on the phospholipid moiety and tissue considered. Finally the direct acylation of lysophospholipids formed during the process of membrane turnover may contribute to both rapid and acclimatory adjustments in membrane lipid composition.

Sign in / Sign up

Export Citation Format

Share Document