scholarly journals Benzoquinone alters the lipid homeostasis in Saccharomyces cerevisiae

2019 ◽  
Vol 8 (6) ◽  
pp. 1035-1041
Author(s):  
Abhishek Raj ◽  
Vasanthi Nachiappan
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Neutral Lipids ◽  
Gene Knockout ◽  
Phospholipid Metabolism ◽  
Lipid Homeostasis ◽  
Wild Type ◽  
Kennedy Pathway ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
The Impact

Abstract Objective: To elucidate the impact of benzoquinone (BQ) on lipid homeostasis and cytotoxicity in Saccharomyces cerevisiae. Methods: The impact of BQ exposure on wild-type and knockouts of PC biosynthesizing genes revealed the alterations in the lipids that were analyzed by fluorescence microscopy, thin layer chromatography, and gene expression studies. Results: In yeast, BQ exposure reduced the growth pattern in wild-type cells. The gene knockout strains of the phospholipid metabolism altered the mRNA expression of the apoptosis genes – both caspase-dependent and independent. The BQ exposure revealed an increase in both the phospholipids and neutral lipids via the CDP:DAG and the Kennedy pathway genes. The accumulation of both neutral lipids and phospholipids during the BQ exposure was discrete and regulated by different pathways. Conclusions: BQ exposure inhibited cell growth, increased the reactive oxygen species (ROS), and altered membrane proliferation. The CDP:DAG and Kennedy pathway lipids also discretely altered by BQ, which is required for the membrane functions and energy purposes of life.

scholarly journals Hydroquinone exposure accumulates neutral lipid by the activation of CDP-DAG pathway in Saccharomyces cerevisiae

2021 ◽  
Author(s):  
Abhishek Raj ◽  
Vasanthi Nachiappan
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Neutral Lipid ◽  
Neutral Lipids ◽  
Model Organism ◽  
Membrane Lipid ◽  
Vital Role ◽  
Cellular Respiration ◽  
Wild Type ◽  
Kennedy Pathway ◽  
In The Wild

Abstract Benzene metabolites (HQ and BQ) are toxic compounds and their presence in human cause alteration in cellular respiration and kidney damage. In the current study, Saccharomyces cerevisiae has been used as a model organism and acute exposure of hydroquinone (HQ) decreased cell growth and increased reactive oxygen species (ROS). The expression of apoptosis regulatory genes (YCA1, NUC1, YSP1 and AIF1) were increased with HQ exposure in the wild-type cells. HQ exposure in the wild-type cells altered both the phospholipid and neutral lipid levels. Phosphatidylcholine is a vital membrane lipid that has a vital role in membrane biogenesis and was increased significantly with HQ. The neutral lipid results were supported with lipid droplets data and mRNA expression study. The phospholipid knockouts (Kennedy pathway) accumulated neutral lipids via the CDP-DAG (cytidine-diphosphate-diacylglycerol) pathway genes both in the presence and absence of HQ.

The chinless mutation and neural crest cell interactions in zebrafish jaw development

Development ◽  
1996 ◽  
Vol 122 (5) ◽  
pp. 1417-1426 ◽  
Author(s):  
T.F. Schilling ◽  
C. Walker ◽  
C.B. Kimmel
Keyword(s):  
Neural Crest ◽  
Neural Crest Cell ◽  
Neural Crest Cells ◽  
Paraxial Mesoderm ◽  
Wild Type ◽  
Host Genotype ◽  
Pharyngeal Arches ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Cranial Neural Crest Cells

During vertebrate development, neural crest cells are thought to pattern many aspects of head organization, including the segmented skeleton and musculature of the jaw and gills. Here we describe mutations at the gene chinless, chn, that disrupt the skeletal fates of neural crest cells in the head of the zebrafish and their interactions with muscle precursors. chn mutants lack neural-crest-derived cartilage and mesoderm-derived muscles in all seven pharyngeal arches. Fate mapping and gene expression studies demonstrate the presence of both undifferentiated cartilage and muscle precursors in mutants. However, chn blocks differentiation directly in neural crest, and not in mesoderm, as revealed by mosaic analyses. Neural crest cells taken from wild-type donor embryos can form cartilage when transplanted into chn mutant hosts and rescue some of the patterning defects of mutant pharyngeal arches. In these cases, cartilage only forms if neural crest is transplanted at least one hour before its migration, suggesting that interactions occur transiently in early jaw precursors. In contrast, transplanted cells in paraxial mesoderm behave according to the host genotype; mutant cells form jaw muscles in a wild-type environment. These results suggest that chn is required for the development of pharyngeal cartilages from cranial neural crest cells and subsequent crest signals that pattern mesodermally derived myocytes.

scholarly journals Large-scale gene expression studies of the endometrium: what have we learnt?

Reproduction ◽  
2006 ◽  
Vol 132 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Rob Sherwin ◽  
Rob Catalano ◽  
Andrew Sharkey
Keyword(s):  
Large Scale ◽  
Dna Microarrays ◽  
Disease States ◽  
Complex Interactions ◽  
Expression Studies ◽  
Proliferation And Differentiation ◽  
Potential Applications ◽  
Underlying Causes ◽  
Gene Expression Studies ◽  
The Impact

The endometrium is a dynamic tissue that undergoes coordinated changes under the influence of steroid hormones. This results in proliferation and differentiation culminating in a receptive state, followed by menstruation and endometrial repair. These functions involve complex interactions between the epithelium, stroma and leucocytes in the endometrium. Understanding the underlying causes of endometrial disorders, such as infertility, endometriosis and heavy menstrual bleeding, therefore represents a considerable challenge. Recently developed techniques, such as differential display and DNA microarrays permit the abundance of thousands of mRNA transcripts within cells or tissues to be measured simultaneously. This provides a new approach to understanding the complex interactions that underlie both healthy and disease states. Responses of the endometrium to hormones or drugs can be studied and the response of the system as an integrated whole can be assessed. Comparisons of endometrium from healthy women and those with endometrial dysfunction have advanced our understanding of key areas of endometrial physiology, including infertility, receptivity, endometriosis and cancer. Using this approach, novel genes controlling specific endometrial functions like receptivity have been identified for functional testing. This paper will review the impact of these techniques for transcript profiling on our understanding of selected areas of endometrial biology and discuss the potential applications in future.

scholarly journals MmpS5-MmpL5 Transporters Provide Mycobacterium smegmatis Resistance to imidazo[1,2-b][1,2,4,5]tetrazines

Pathogens ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 166 ◽  
Author(s):  
Dmitry A. Maslov ◽  
Kirill V. Shur ◽  
Aleksey A. Vatlin ◽  
Valery N. Danilenko
Keyword(s):  
Gene Expression ◽  
Mycobacterium Smegmatis ◽  
Drug Resistant ◽  
Wild Type ◽  
Efflux System ◽  
Mechanisms Of Resistance ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Mutant Genes

The emergence and spread of drug-resistant Mycobacterium tuberculosis strains (including MDR, XDR, and TDR) force scientists worldwide to search for new anti-tuberculosis drugs. We have previously reported a number of imidazo[1,2-b][1,2,4,5]tetrazines – putative inhibitors of mycobacterial eukaryotic-type serine-threonine protein-kinases, active against M. tuberculosis. Whole genomic sequences of spontaneous drug-resistant M. smegmatis mutants revealed four genes possibly involved in imidazo[1,2-b][1,2,4,5]tetrazines resistance; however, the exact mechanism of resistance remain unknown. We used different approaches (construction of targeted mutants, overexpression of the wild-type (w.t.) and mutant genes, and gene-expression studies) to assess the role of the previously identified mutations. We show that mutations in MSMEG_1380 gene lead to overexpression of the mmpS5-mmpL5 operon in M. smegmatis, thus providing resistance to imidazo[1,2-b][1,2,4,5]tetrazines by increased efflux through the MmpS5-MmpL5 system, similarly to the mechanisms of resistance described for M. tuberculosis and M. abscessus. Mycobacterial MmpS5-MmpL5 transporters should be considered as an MDR-efflux system and they should be taken into account at early stages of anti-tuberculosis drug development.

scholarly journals σB Activation under Environmental and Energy Stress Conditions in Listeria monocytogenes

2006 ◽  
Vol 72 (8) ◽  
pp. 5197-5203 ◽  
Author(s):  
Soraya Chaturongakul ◽  
Kathryn J. Boor
Keyword(s):  
Listeria Monocytogenes ◽  
Cumene Hydroperoxide ◽  
Stress Conditions ◽  
Wild Type ◽  
Regulation Of Expression ◽  
Transcript Levels ◽  
Energy Stress ◽  
Expression Studies ◽  
Genes Encoding ◽  
Gene Expression Studies

ABSTRACT To measure σB activation in Listeria monocytogenes under environmental or energy stress conditions, quantitative reverse transcriptase PCR (TaqMan) was used to determine the levels of transcripts for the σB-dependent opuCA and clpC genes in strains having null mutations in genes encoding regulator of sigma B proteins (rsbT and rsbV) and sigma B (sigB) and in the L. monocytogenes wild-type 10403S strain under different stress conditions. The ΔsigB, ΔrsbT, and ΔrsbV strains previously exhibited increased hemolytic activities compared to the hemolytic activity of the wild-type strain; therefore, transcript levels for hly were also determined. RsbT, RsbV, and σB were all required for opuCA expression during growth under carbon-limiting conditions or following exposure to pH 4.5, salt, ethanol, or the protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP). Expression of clpC was RsbT, RsbV, and σB dependent in the presence of CCCP but not under the other conditions. hly expression was not RsbT, RsbV, or σB dependent in the presence of either CCCP or salt. opuCA transcript levels did not increase in the presence of rapidly lethal stresses (i.e., pH 2.5 or 13 mM cumene hydroperoxide) despite the enhanced survival of the wild type compared with the survival of the mutant strains under these conditions. These findings highlight the importance of complementing phenotypic characterizations with gene expression studies to identify direct and indirect effects of null mutations in regulatory genes, such as sigB. Overall, our data show that while σB activation occurs through a single pathway under both environmental and energy stress conditions, regulation of expression of some stress response and virulence genes in the σB regulon (e.g., clpC) appears to require networks involving multiple transcriptional regulators.

scholarly journals Inositol and Phosphate Regulate GIT1 Transcription and Glycerophosphoinositol Incorporation in Saccharomyces cerevisiae

2003 ◽  
Vol 2 (4) ◽  
pp. 729-736 ◽  
Author(s):  
C. Almaguer ◽  
D. Mantella ◽  
E. Perez ◽  
J. Patton-Vogt
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasma Membrane ◽  
Wild Type Strain ◽  
Phospholipid Metabolism ◽  
Phosphate Metabolism ◽  
Wild Type ◽  
Factors Affecting ◽  
Low Concentrations ◽  
Free Media ◽  
Regulatory Connection

ABSTRACT Glycerophosphoinositol is produced through deacylation of the essential phospholipid phosphatidylinositol. In Saccharomyces cerevisiae, the glycerophosphoinositol produced is excreted from the cell but is recycled for phosphatidylinositol synthesis when inositol is limiting. To be recycled, glycerophosphoinositol enters the cell through the permease encoded by GIT1. The transport of exogenous glycerophosphoinositol through Git1p is sufficiently robust to support the growth of an inositol auxotroph (ino1Δ). We now report that S. cerevisiae also uses exogenous phosphatidylinositol as an inositol source. Evidence suggests that phosphatidylinositol is deacylated to glycerophosphoinositol extracellularly before being transported across the plasma membrane by Git1p. A genetic screen identified Pho86p, which is required for targeting of the major phosphate transporter (Pho84p) to the plasma membrane, as affecting the utilization of phosphatidylinositol and glycerophosphoinositol. Deletion of PHO86 in an ino1Δ strain resulted in faster growth when either phosphatidylinositol or glycerophosphoinositol was supplied as the sole inositol source. The incorporation of radiolabeled glycerophosphoinositol into an ino1Δ pho86Δ mutant was higher than that into wild-type, ino1Δ, and pho86Δ strains. All strains accumulated the most GIT1 transcript when incubated in media limited for inositol and phosphate in combination. However, the ino1Δ pho86Δ mutant accumulated approximately threefold more GIT1 transcript than did the other strains when incubated in inositol-free media containing either high or low concentrations of Pi. Deletion of PHO4 abolished GIT1 transcription in a wild-type strain. These results indicate that the transport of glycerophosphoinositol by Git1p is regulated by factors affecting both inositol and phosphate availabilities and suggest a regulatory connection between phosphate metabolism and phospholipid metabolism.

scholarly journals Arabidopsis ERF4 and MYB52 Transcription Factors Play Antagonistic Roles in Regulating Homogalacturonan De-methylesterification in Seed Coat Mucilage

2020 ◽  
Author(s):  
Anming Ding ◽  
Xianfeng Tang ◽  
Linhe Han ◽  
Jianlu Sun ◽  
Angyan Ren ◽  
...  
Keyword(s):  
Cell Wall ◽  
Transcription Factors ◽  
Seed Coat ◽  
Pectin Methylesterase ◽  
Wild Type ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Transcriptional Complex ◽  
Seed Coat Mucilage ◽  
Seed Mucilage

ABSTRACTThe Arabidopsis (Arabidopsis thaliana) seed coat mucilage is a specialized cell wall with pectin as its major component. Pectin is synthesized in the Golgi apparatus with homogalacturonan fully methylesterified, but it must undergo de-methylesterification by pectin methylesterase (PME) after being secreted into the cell wall. This reaction is critical for pectin maturation, but the mechanisms of its transcriptional regulation remain largely unknown. Here, we show that the Arabidopsis ERF4 transcription factor positively regulates pectin de-methylesterification during seed development and directly suppresses the expression of PME INHIBITOR13 (PMEI13), 14, 15 and SUBTILISIN-LIKE SERINE PROTEASE 1.7 (SBT1.7). The erf4 mutant seeds showed repartitioning of mucilage between soluble and adherent layers as a result of decreased PME activity and increased degree of pectin methylesterification. ERF4 physically associates with and antagonizes MYB52 in activating PMEI6, 14 and SBT1.7 and MYB52 also antagonizes ERF4 activity in the regulation of downstream targets. Gene expression studies revealed that ERF4 and MYB52 have opposite effects on pectin de-methylesterification. Genetic analysis indicated that the erf4-2 myb52 double mutant seeds show mucilage phenotype similar to wild-type. Taken together, this study demonstrates that ERF4 and MYB52 antagonize each other’s activity to maintain the appropriate degree of pectin methylesterification, expanding our understanding of how pectin de-methylesterification is fine-tuned by the ERF4-MYB52 transcriptional complex in the seed mucilage.One-sentence summaryArabidopsis ERF4 and MYB52 transcription factors interact and play antagonistic roles in regulating homogalacturonan de-methylesterification related genes in the seed coat mucilage.

scholarly journals Analysis of the Contribution of cyp51 Genes to Azole Resistance in Aspergillus Section Nigri with the CRISPR-Cas9 Technique

2021 ◽  
Vol 65 (5) ◽  
Author(s):  
Alba Pérez-Cantero ◽  
Adela Martin-Vicente ◽  
Josep Guarro ◽  
Jarrod R. Fortwendel ◽  
Javier Capilla
Keyword(s):  
Single Gene ◽  
Transcriptional Response ◽  
Azole Resistance ◽  
Gene Deletions ◽  
Content Type ◽  
Ergosterol Content ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Induced Changes ◽  
The Impact

ABSTRACT Cyp51 contribution to azole resistance has been broadly studied and characterized in Aspergillus fumigatus, whereas it remains poorly investigated in other clinically relevant species of the genus, such as those of section Nigri. In this work, we aimed to analyze the impact of cyp51 genes (cyp51A and cyp51B) on the voriconazole (VRC) response and resistance of Aspergillus niger and Aspergillus tubingensis. We generated CRISPR-Cas9 cyp51A and cyp51B knockout mutants from strains with different genetic backgrounds and diverse patterns of azole susceptibility. Single-gene deletions of cyp51 genes resulted in 2- to 16-fold decreases of the VRC MIC values, which were below the VRC epidemiological cutoff value (ECV) established by the Clinical and Laboratory Standards Institute (CLSI), irrespective of their parental strains’ susceptibilities. Gene expression studies in the tested species confirmed that cyp51A participates more actively than cyp51B in the transcriptional response of azole stress. However, ergosterol quantification revealed that both enzymes comparably impact the total ergosterol content within the cell, as basal- and VRC-induced changes to ergosterol content were similar in all cases. These data contribute to our understanding of Aspergillus azole resistance, especially in non-A. fumigatus species.

scholarly journals Analysis of biosynthesis and composition of cuticular wax in wild type bilberry (Vaccinium myrtillus L.) and its glossy mutant

2020 ◽  
Author(s):  
Priyanka Trivedi ◽  
Nga Nguyen ◽  
Linards Klavins ◽  
Jorens Kviesis ◽  
Esa Heinonen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Chemical Composition ◽  
Fruit Development ◽  
Cuticular Wax ◽  
Wild Type ◽  
Expression Studies ◽  
Fruit Skin ◽  
Type Mutant ◽  
Gene Expression Studies ◽  
Epicuticular Wax Layer

AbstractCuticular wax plays an important role in fruits in protection against environmental stresses and desiccation. In this study, biosynthesis and chemical composition of cuticular wax in wild type (WT) bilberry fruit was studied during development and compared with its natural glossy type (GT) mutant. The cuticular wax load in GT fruit was comparable to WT fruit. In both fruits, triterpenoids were the dominant wax compounds with decreasing proportion during the fruit development accompanied with increasing proportion of aliphatic compounds. Gene expression studies supported the pattern of compound accumulation during fruit development. Genes CER26-like, FAR2, CER3-like, LTP, MIXTA, and BAS exhibited prevalent expression in fruit skin indicating role in cuticular wax biosynthesis and secretion. In GT fruit, higher proportion of triterpenoids in cuticular wax was accompanied by lower proportion of fatty acids and ketones compared to WT fruit as well as lower density of crystalloid structures on berry surface. Our results suggest that a marked reduction in ketones in cuticular wax may play a significant role in the formation of glossy phenotype leading to the loss of rod-like structures in epicuticular wax layer of GT fruit.HighlightChemical composition and morphology of cuticular wax along with gene expression for wax biosynthetic genes varied between glossy type mutant (GT) and wild type (WT) fruit.

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