scholarly journals Proteome Analysis of Streptococcus thermophilus Grown in Milk Reveals Pyruvate Formate-Lyase as the Major Upregulated Protein

2005 ◽  
Vol 71 (12) ◽  
pp. 8597-8605 ◽  
Author(s):  
Sylviane Derzelle ◽  
Alexander Bolotin ◽  
Michel-Yves Mistou ◽  
Françoise Rul
Keyword(s):  
Anaerobic Fermentation ◽  
Physiological Role ◽  
Proteome Analysis ◽  
Streptococcus Thermophilus ◽  
Product Formation ◽  
Exponential Growth Phase ◽  
Transcriptional Level ◽  
Acid Product ◽  
Pyruvate Formate Lyase ◽  
Proteomic Approach

ABSTRACT We investigated the adaptation to milk of Streptococcus thermophilus LMG18311 using a proteomic approach. Two-dimensional electrophoresis of cytosolic proteins were performed after growth in M17 medium or in milk. A major modification of the proteome concerned proteins involved in the supply of amino acids, like the peptidase PepX, and several enzymes involved in amino acid biosynthesis. In parallel, we observed the upregulation of the synthesis of seven enzymes directly involved in the synthesis of purines, as well as formyl-tetrahydrofolate (THF) synthetase and serine hydroxy-methyl transferase, two enzymes responsible for the synthesis of compounds (THF and glycine, respectively) feeding the purine biosynthetic pathway. The analysis also revealed a massive increase in the synthesis of pyruvate formate-lyase (PFL), the enzyme which converts pyruvate into acetyl coenzyme A and formate. PFL has been essentially studied for its role in mixed-acid product formation in lactic acid bacteria during anaerobic fermentation. However, formate is an important methyl group donor for anabolic pathway through the formation of folate derivates. We hypothesized that PFL was involved in purine biosynthesis during growth in milk. We showed that PFL expression was regulated at the transcriptional level and that pfl transcription occurred during the exponential growth phase in milk. The complementation of milk with formate or purine bases was shown to reduce pfl expression, to suppress PFL synthesis, and to stimulate growth of S. thermophilus. These results show a novel regulatory mechanism controlling the synthesis of PFL and suggest an unrecognized physiological role for PFL as a formate supplier for anabolic purposes.

scholarly journals p38/AP-1 Pathway in Lipopolysaccharide-Induced Inflammatory Responses Is Negatively Modulated by Electrical Stimulation

2013 ◽  
Vol 2013 ◽  
pp. 1-11 ◽  
Author(s):  
Deok Jeong ◽  
Jaehwi Lee ◽  
Young-Su Yi ◽  
Yanyan Yang ◽  
Kyoung Won Kim ◽  
...  
Keyword(s):  
Electrical Stimulation ◽  
Inflammatory Response ◽  
Inflammatory Responses ◽  
Inflammatory Diseases ◽  
Physiological Role ◽  
Peritoneal Macrophages ◽  
Cellular Level ◽  
Transcriptional Level ◽  
Weak Current ◽  
Anti Inflammatory

Electrical stimulation with a weak current has been demonstrated to modulate various cellular and physiological responses, including the differentiation of mesenchymal stem cells and acute or chronic physical pain. Thus, a variety of investigations regarding the physiological role of nano- or microlevel currents at the cellular level are actively proceeding in the field of alternative medicine. In this study, we focused on the anti-inflammatory activity of aluminum-copper patches (ACPs) under macrophage-mediated inflammatory conditions. ACPs generated nanolevel currents ranging from 30 to 55 nA in solution conditions. Interestingly, the nanocurrent-generating aluminum-copper patches (NGACPs) were able to suppress both lipopolysaccharide-(LPS-) and pam3CSK-induced inflammatory responses such as NO and PGE2production in both RAW264.7 cells and peritoneal macrophages at the transcriptional level. Through immunoblotting and immunoprecipitation analyses, we found that p38/AP-1 could be the major inhibitory pathway in the NGACP-mediated anti-inflammatory response. Indeed, inhibition of p38 by SB203580 showed similar inhibitory activity of the production of TNF-αand PGE2and the expression of TNF-αand COX-2 mRNA. These results suggest that ACP-induced nanocurrents alter signal transduction pathways that are involved in the inflammatory response and could therefore be utilized in the treatment of various inflammatory diseases such as arthritis and colitis.

Comparative whey proteome analysis of small-tailed Han and DairyMeade ovine milk

2021 ◽  
pp. 1-5
Author(s):  
Urhan Bai ◽  
Xiaohu Su ◽  
Zhong Zheng ◽  
Liguo Zhang ◽  
Ying Ma ◽  
...  
Keyword(s):  
Mammary Gland Development ◽  
Proteome Analysis ◽  
Dairy Sheep ◽  
Milk Traits ◽  
Beneficial Effects ◽  
Proteomic Approach ◽  
Tandem Mass Tag ◽  
Proteome Profile ◽  
Related Proteins ◽  
Gland Development

Abstract We characterized the proteome profile of mid-lactation small-tailed Han (STH) and DairyMeade (DM) ovine milk in order to explore physiological variation and differences in milk traits between the two breeds. Methodology combined a tandem mass tag (TMT) proteomic approach with LC-MS/MS technology. A total of 656 proteins were identified in STH and DM ovine milk, of which 17and 29 proteins were significantly upregulated (P < 0.05) in STH and DM, respectively. Immune-related proteins and disease-related proteins were highly expressed in STH milk, whereas S100A2 and AEBP1 were highly expressed in DM milk, which had beneficial effects on mammary gland development and milk yield. Our results provide a theoretical basis for future breeding of dairy sheep.

scholarly journals Regulatory Mechanism of MicroRNA Expression in Cancer

2020 ◽  
Vol 21 (5) ◽  
pp. 1723 ◽  
Author(s):  
Zainab Ali Syeda ◽  
Siu Semar Saratu’ Langden ◽  
Choijamts Munkhzul ◽  
Mihye Lee ◽  
Su Jung Song
Keyword(s):  
Molecular Mechanism ◽  
Mirna Expression ◽  
Messenger Rna ◽  
Target Genes ◽  
Regulatory Mechanism ◽  
Physiological Role ◽  
Mirna Biogenesis ◽  
Transcriptional Level ◽  
Detailed Knowledge ◽  
Cancer Pathogenesis

Altered gene expression is the primary molecular mechanism responsible for the pathological processes of human diseases, including cancer. MicroRNAs (miRNAs) are virtually involved at the post-transcriptional level and bind to 3′ UTR of their target messenger RNA (mRNA) to suppress expression. Dysfunction of miRNAs disturbs expression of oncogenic or tumor-suppressive target genes, which is implicated in cancer pathogenesis. As such, a large number of miRNAs have been found to be downregulated or upregulated in human cancers and to function as oncomiRs or oncosuppressor miRs. Notably, the molecular mechanism underlying the dysregulation of miRNA expression in cancer has been recently uncovered. The genetic deletion or amplification and epigenetic methylation of miRNA genomic loci and the transcription factor-mediated regulation of primary miRNA often alter the landscape of miRNA expression in cancer. Dysregulation of the multiple processing steps in mature miRNA biogenesis can also cause alterations in miRNA expression in cancer. Detailed knowledge of the regulatory mechanism of miRNAs in cancer is essential for understanding its physiological role and the implications of cancer-associated dysfunction and dysregulation. In this review, we elucidate how miRNA expression is deregulated in cancer, paying particular attention to the cancer-associated transcriptional and post-transcriptional factors that execute miRNA programs.

Study of pH Effect on the Anaerobic-Aerobic Fermentation of Siwalan (Borassus flabellifer L.) Sap to Produce Acetic Acid

2019 ◽  
Vol 964 ◽  
pp. 209-214
Author(s):  
Elly Agustiani ◽  
Atiqa Rahmawati ◽  
Fibrillian Zata Lini ◽  
Dimas Luthfi Ramadhani
Keyword(s):  
Acetic Acid ◽  
Acid Concentration ◽  
Ethanol Fermentation ◽  
Anaerobic Fermentation ◽  
Sugar Content ◽  
Alcoholic Beverages ◽  
Acetic Acid Concentration ◽  
Acetobacter Aceti ◽  
Aerobic Fermentation ◽  
Acid Product

Siwalan (Borassus flabellifer L.) is a palm family that is widely planted in the Tuban area of ​​East Java. Siwalan sap has a relatively high sugar content of about 10-15 g / 100 ml. The sap is obtained by tapping the inflorescences. In general, siwalan sap is used for fresh drinks or alcoholic beverages with maximum storage in 3 days. Based on the sugar content in the sap of siwalan, acetic acid products can be made through fermentation of glucose to ethanol, then the ethanol is fermented into acetic acid. Acetic acid is widely used as a preservative of food and health drinks. The purpose of this research is to study the effect of ethanol fermentation aerobic pH on acetic acid product. Anaerobic fermentation uses saccharomyces cereviceae to produce ethanol, and aerobic fermentation uses acetobacter aceti for acetic acid production. In aerobic ethanol fermentation using pH 3; 3.5; 4 and 5. The concentration of ethanol was analyzed using GC ULTRA Scientific Gas Chromatography, DSQ II detector, and MS 220 column. Acetic acid produced from the aerobic fermentation process was analyzed using an alkalimetric method. Anaerobic fermentation uses Saccharomyces cereviceae with 1-day log phase, while aerobic fermentation uses acetobacter aceti with a 5 day log phase. Aerobic fermentation to produce acetic acid was observed in 5 days to obtained maximum acetic acid concentration, the highest acetic acid concetration is about 2.595 g/l and yield of acetic acid is obtained 0.519% (b/v) at pH 5. Low acetic acid concentration due to low intitial sugar content in siwalan sap.

scholarly journals Insights into Regulation and Function of the Major Stress-Induced hsp70 Molecular Chaperone In Vivo: Analysis of Mice with Targeted Gene Disruption of the hsp70.1 orhsp70.3 Gene

2001 ◽  
Vol 21 (24) ◽  
pp. 8575-8591 ◽  
Author(s):  
Lei Huang ◽  
Nahid F. Mivechi ◽  
Demetrius Moskophidis
Keyword(s):  
Heat Stress ◽  
Gene Family ◽  
Synergistic Effects ◽  
Single Gene ◽  
Physiological Role ◽  
Evolutionary Significance ◽  
Homologous Gene ◽  
Transcriptional Level ◽  
Adult Mice

ABSTRACT The murine hsp70 gene family includes the evolutionarily conserved hsp70.1 andhsp70.3 genes, which are the major proteins induced by heat and other stress stimuli.hsp70.1 andhsp70.3 encode identical proteins which protect cells and facilitate their recovery from stress-induced damage. While the hsp70 gene family has been widely studied and the roles of the proteins it encodes as molecular chaperones in a range of human pathologies are appreciated, little is known about the developmental regulation of hsp70.1 andhsp70.3 expression and the in vivo biological function of their products. To directly study the physiological role of these proteins in vivo, we have generated mice deficient in heat shock protein 70 (hsp70) by replacing thehsp70.1 orhsp70.3 gene with an in-frame β-galactosidase sequence. We report here that the expression ofhsp70.1 andhsp70.3 is developmentally regulated at the transcriptional level, and an overlapping expression pattern for both genes is observed during embryo development and in the tissues of adult mice. hsp70.1 −/− orhsp70.3 −/− mice are viable and fertile, with no obvious morphological abnormalities. In late embryonic stage and adult mice, both genes are expressed constitutively in tissues exposed directly to the environment (the epidermis and cornea) and in certain internal organs (the epithelium of the tongue, esophagus, and forestomach, and the kidney, bladder, and hippocampus). Exposure of mice to thermal stress results in the rapid induction and expression of hsp70, especially in organs not constitutively expressing hsp70 (the liver, pancreas, heart, lung, adrenal cortex, and intestine). Despite functional compensation in the single-gene-deficient mice by the intact homologous gene (i.e.,hsp70.3 inhsp70.1 −/− mice and vice versa), a marked reduction in hsp70 protein expression was observed in tissues under both normal and heat stress conditions. At the cellular level, inactivation of hsp70.1 orhsp70.3 resulted in deficient maintenance of acquired thermotolerance and increased sensitivity to heat stress-induced apoptosis. The additive or synergistic effects exhibited by coexpression of both hsp70 genes, and the evolutionary significance of the presence of both hsp70genes, is hence underlined.

scholarly journals The Oligopeptide Transport System Is Essential for the Development of Natural Competence in Streptococcus thermophilus Strain LMD-9

2009 ◽  
Vol 191 (14) ◽  
pp. 4647-4655 ◽  
Author(s):  
Rozenn Gardan ◽  
Colette Besset ◽  
Alain Guillot ◽  
Christophe Gitton ◽  
Véronique Monnet
Keyword(s):  
Quorum Sensing ◽  
Streptococcus Thermophilus ◽  
Defined Medium ◽  
Mass Spectrometry Analysis ◽  
Transport Systems ◽  
Label Free ◽  
Spectrometry Analysis ◽  
Proteomic Approach ◽  
Liquid Chromatography Tandem Mass ◽  
Oligopeptide Transport

ABSTRACT In gram-positive bacteria, oligopeptide transport systems, called Opp or Ami, play a role in nutrition but are also involved in the internalization of signaling peptides that take part in the functioning of quorum-sensing pathways. Our objective was to reveal functions that are controlled by Ami via quorum-sensing mechanisms in Streptococcus thermophilus, a nonpathogenic bacterium widely used in dairy technology in association with other bacteria. Using a label-free proteomic approach combining one-dimensional electrophoresis with liquid chromatography-tandem mass spectrometry analysis, we compared the proteome of the S. thermophilus LMD-9 to that of a mutant deleted for the subunits C, D, and E of the ami operon. Both strains were grown in a chemically defined medium (CDM) without peptides. We focused our attention on proteins that were no more detected in the ami deletion mutant. In addition to the three subunits of the Ami transporter, 17 proteins fulfilled this criterion and, among them, 7 were similar to proteins that have been identified as essential for transformation in S. pneumoniae. These results led us to find a condition of growth, the early exponential state in CDM, that allows natural transformation in S. thermophilus LMD-9 to turn on spontaneously. Cells were not competent in M17 rich medium. Furthermore, we demonstrated that the Ami transporter controls the triggering of the competence state through the control of the transcription of comX, itself controlling the transcription of late competence genes. We also showed that one of the two oligopeptide-binding proteins of strain LMD-9 plays the predominant role in the control of competence.

scholarly journals Regulation of gbpC Expression in Streptococcus mutans

2007 ◽  
Vol 189 (18) ◽  
pp. 6521-6531 ◽  
Author(s):  
Indranil Biswas ◽  
Laura Drake ◽  
Saswati Biswas
Keyword(s):  
Streptococcus Mutans ◽  
Dna Sequences ◽  
Promoter Region ◽  
Response Regulator ◽  
Surface Protein ◽  
Maximum Level ◽  
Stress Conditions ◽  
Pcr Analysis ◽  
Exponential Growth Phase ◽  
Transcriptional Level

ABSTRACT Streptococcus mutans, the principal causative agent of dental caries, produces four glucan-binding proteins (Gbp) that play major roles in bacterial adherence and pathogenesis. One of these proteins, GbpC, is an important cell surface protein involved in biofilm formation. GbpC is also important for cariogenesis, bacteremia, and infective endocarditis. In this study, we examined the regulation of gbpC expression in S. mutans strain UA159. We found that gbpC expression attains the maximum level at mid-exponential growth phase, and the half-life of the transcript is less than 2 min. Expression from PgbpC was measured using a PgbpC-gusA transcriptional fusion reporter and was analyzed under various stress conditions, including thermal, osmotic, and acid stresses. Expression of gbpC is induced under conditions of thermal stress but is repressed during growth at low pH, whereas osmotic stress had no effect on expression from PgbpC. The results from the expression analyses were further confirmed using semiquantitative reverse transcription-PCR analysis. Our results also reveal that CovR, a global response regulator in many Streptococcus spp., represses gbpC expression at the transcriptional level. We demonstrated that purified CovR protein binds directly to the promoter region of PgbpC to repress gbpC expression. Using a DNase I protection assay, we showed that CovR binds to DNA sequences surrounding PgbpC from bases −68 to 28 (where base 1 is the start of transcription). In summary, our results indicate that various stress conditions modulate the expression of gbpC and that CovR negatively regulates the expression of the gbpC gene by directly binding to the promoter region.

Oxidative stress-related responses of Bifidobacterium longum subsp. longum BBMN68 at the proteomic level after exposure to oxygen

Microbiology ◽  
2011 ◽  
Vol 157 (6) ◽  
pp. 1573-1588 ◽  
Author(s):  
Man Xiao ◽  
Pan Xu ◽  
Jianyun Zhao ◽  
Zeng Wang ◽  
Fanglei Zuo ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Oxidative Damage ◽  
Bifidobacterium Longum ◽  
Transcriptional Level ◽  
Low Oxygen ◽  
Oxygen Stress ◽  
Dna Oxidative Damage ◽  
Proteomic Approach ◽  
Protective Proteins

Bifidobacterium longum subsp. longum BBMN68, an anaerobic probiotic isolated from healthy centenarian faeces, shows low oxygen (3 %, v/v) tolerance. To understand the effects of oxidative stress and the mechanisms protecting against it in this strain, a proteomic approach was taken to analyse changes in the cellular protein profiles of BBMN68 under the following oxygen-stress conditions. Mid-exponential phase BBMN68 cells grown in MRS broth at 37 °C were exposed to 3 % O2 for 1 h (I) or 9 h (II), and stationary phase cells were subjected to 3 % O2 for 1 h (III). Respective controls were grown under identical conditions but were not exposed to O2. A total of 51 spots with significant changes after exposure to oxygen were identified, including the oxidative stress-protective proteins alkyl hydroperoxide reductase C22 (AhpC) and pyridine nucleotide-disulfide reductase (PNDR), and the DNA oxidative damage-protective proteins DNA-binding ferritin-like protein (Dps), ribonucleotide reductase (NrdA) and nucleotide triphosphate (NTP) pyrophosphohydrolases (MutT1). Changes in polynucleotide phosphorylase (PNPase) plus enolase, which may play important roles in scavenging oxidatively damaged RNA, were also found. Following validation at the transcriptional level of differentially expressed proteins, the physiological and biochemical functions of BBMN68 Dps were further proven by in vitro and in vivo tests under oxidative stress. Our results reveal the key oxidative stress-protective proteins and DNA oxidative damage-protective proteins involved in the defence strategy of BBMN68 against oxygen, and provide the first proteomic information toward understanding the responses of Bifidobacterium and other anaerobes to oxygen stress.

scholarly journals Adaptation of Bacillus subtilis to growth at low temperature: a combined transcriptomic and proteomic appraisal

Microbiology ◽  
2006 ◽  
Vol 152 (3) ◽  
pp. 831-853 ◽  
Author(s):  
Ina Budde ◽  
Leif Steil ◽  
Christian Scharf ◽  
Uwe Völker ◽  
Erhard Bremer
Keyword(s):  
Bacillus Subtilis ◽  
Low Temperature ◽  
Response Regulator ◽  
Proteome Analysis ◽  
Atp Synthesis ◽  
Fine Tuning ◽  
Transcriptional Responses ◽  
Natural Habitats ◽  
Regulatory Circuit ◽  
Proteomic Approach

The soil bacterium Bacillus subtilis frequently encounters a reduction in temperature in its natural habitats. Here, a combined transcriptomic and proteomic approach has been used to analyse the adaptational responses of B. subtilis to low temperature. Propagation of B. subtilis in minimal medium at 15 °C triggered the induction of 279 genes and the repression of 301 genes in comparison to cells grown at 37 °C. The analysis thus revealed profound adjustments in the overall gene expression profile in chill-adapted cells. Important transcriptional changes in low-temperature-grown cells comprise the induction of the SigB-controlled general stress regulon, the induction of parts of the early sporulation regulons (SigF, SigE and SigG) and the induction of a regulatory circuit (RapA/PhrA and Opp) that is involved in the fine-tuning of the phosphorylation status of the Spo0A response regulator. The analysis of chill-stress-repressed genes revealed reductions in major catabolic (glycolysis, oxidative phosphorylation, ATP synthesis) and anabolic routes (biosynthesis of purines, pyrimidines, haem and fatty acids) that likely reflect the slower growth rates at low temperature. Low-temperature repression of part of the SigW regulon and of many genes with predicted functions in chemotaxis and motility was also noted. The proteome analysis of chill-adapted cells indicates a major contribution of post-transcriptional regulation phenomena in adaptation to low temperature. Comparative analysis of the previously reported transcriptional responses of cold-shocked B. subtilis cells with this data revealed that cold shock and growth in the cold constitute physiologically distinct phases of the adaptation of B. subtilis to low temperature.

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