scholarly journals Mycoplasma and host interaction: In vitro gene expression modulation in Mycoplasma synoviae and infected chicken chondrocytes

2016 ◽  
Vol 64 (1) ◽  
pp. 26-37 ◽  
Author(s):  
Ivanka Cizelj ◽  
Daliborka Dušanić ◽  
Dušan Benčina ◽  
Mojca Narat
Keyword(s):  
Gene Expression ◽  
Arginine Deiminase ◽  
Gene Encoding ◽  
Mycoplasma Synoviae ◽  
Host Interaction ◽  
Expression Of Genes ◽  
Cartilage Extracellular Matrix ◽  
Genes Encoding ◽  
First Time

The complex interplay between Mycoplasma synoviae and chicken chondrocytes (CCH), which come into direct contact during infectious synovitis, has been examined at the level of gene expression. Our previous studies demonstrated a significant influence of M. synoviae on the level of CCH gene expression. Here, we show for the first time that in vitro co-cultivation of M. synoviae and CCH also induces upregulation of gene expression in this mycoplasma. We observed significantly increased expression of genes important for M. synoviae pathogenicity, including cysteine protease cysP, neuraminidase nanH, haemagglutinin vlhA, and the putative nuclease MS53_0284. Moreover, the pattern of gene expression was dependent on the infection environment. In CCH, significant changes in the expression of genes encoding catabolic enzymes of the cartilage extracellular matrix (cathepsins B, K and L, aggrecanase ADAM10, and matrix metalloproteinase MMP2) were demonstrated. Infection of CCH with M. synoviae also elevated the expression of the gene encoding peptidyl arginine deiminase, type III (PADI3), which is responsible for the post-translational citrullination of proteins.

scholarly journals Nrg1 Is a Transcriptional Repressor for Glucose Repression of STA1 Gene Expression inSaccharomyces cerevisiae

1999 ◽  
Vol 19 (3) ◽  
pp. 2044-2050 ◽  
Author(s):  
Seok Hee Park ◽  
Sang Seok Koh ◽  
Jae Hwan Chun ◽  
Hye Jin Hwang ◽  
Hyen Sam Kang
Keyword(s):  
Gene Expression ◽  
Zinc Finger Protein ◽  
Glucose Repression ◽  
Transcriptional Repressor ◽  
Dependent Manner ◽  
Expression Of Genes ◽  
Dnase I Footprinting ◽  
Genes Encoding

ABSTRACT Expression of genes encoding starch-degrading enzymes is regulated by glucose repression in the yeast Saccharomyces cerevisiae. We have identified a transcriptional repressor, Nrg1, in a genetic screen designed to reveal negative factors involved in the expression of STA1, which encodes a glucoamylase. TheNRG1 gene encodes a 25-kDa C2H2zinc finger protein which specifically binds to two regions in the upstream activation sequence of the STA1 gene, as judged by gel retardation and DNase I footprinting analyses. Disruption of theNRG1 gene causes a fivefold increase in the level of theSTA1 transcript in the presence of glucose. The expression of NRG1 itself is inhibited in the absence of glucose. DNA-bound LexA-Nrg1 represses transcription of a target gene 10.7-fold in a glucose-dependent manner, and this repression is abolished in bothssn6 and tup1 mutants. Two-hybrid and glutathione S-transferase pull-down experiments show an interaction of Nrg1 with Ssn6 both in vivo and in vitro. These findings indicate that Nrg1 acts as a DNA-binding repressor and mediates glucose repression of the STA1 gene expression by recruiting the Ssn6-Tup1 complex.

scholarly journals Drought Stress Enhances Up-Regulation of Anthocyanin Biosynthesis in Grapevine leafroll-associated virus 3-Infected in vitro Grapevine (Vitis vinifera) Leaves

Plant Disease ◽  
2017 ◽  
Vol 101 (9) ◽  
pp. 1606-1615 ◽  
Author(s):  
Zhen-Hua Cui ◽  
Wen-Lu Bi ◽  
Xin-Yi Hao ◽  
Peng-Min Li ◽  
Ying Duan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Drought Stress ◽  
Vitis Vinifera ◽  
Anthocyanin Biosynthesis ◽  
Anthocyanin Accumulation ◽  
Expression Levels ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Regulated Gene Expression

Reddish-purple coloration on the leaf blades and downward rolling of leaf margins are typical symptoms of grapevine leafroll disease (GLD) in red-fruited grapevine cultivars. These typical symptoms are attributed to the expression of genes encoding enzymes for anthocyanins synthesis, and the accumulation of flavonoids in diseased leaves. Drought has been proven to accelerate development of GLD symptoms in virus-infected leaves of grapevine. However, it is not known how drought affects GLD expression nor how anthocyanin biosynthesis in virus-infected leaves is altered. The present study used HPLC to determine the types and levels of anthocyanins, and applied reverse transcription quantitative polymerase chain reaction (RT-qPCR) to analyze the expression of genes encoding enzymes for anthocyanin synthesis. Plantlets of Grapevine leafroll-associated virus 3 (GLRaV-3)-infected Vitis vinifera ‘Cabernet Sauvignon’ were grown in vitro under PEG-induced drought stress. HPLC found no anthocyanin-related peaks in the healthy plantlets with or without PEG-induced stress, while 11 peaks were detected in the infected plantlets with or without PEG-induced drought stress, but the peaks were significantly higher in infected drought-stressed plantlets. Increased accumulation of total anthocyanin compounds was related to the development of GLD symptoms in the infected plantlets under PEG stress. The highest level of up-regulated gene expression was found in GLRaV-3-infected leaves with PEG-induced drought stress. Analyses of variance and correlation of anthocyanin accumulation with related gene expression levels found that GLRaV-3-infection was the key factor in increased anthocyanin accumulation. This accumulation involved the up-regulation of two key genes, MYBA1 and UFGT, and their expression levels were further enhanced by drought stress.

scholarly journals Krüppel-like factor 6–mediated loss of BCAA catabolism contributes to kidney injury in mice and humans

2021 ◽  
Vol 118 (23) ◽  
pp. e2024414118
Author(s):  
Sian E. Piret ◽  
Yiqing Guo ◽  
Ahmed A. Attallah ◽  
Sylvia J. Horne ◽  
Amy Zollman ◽  
...  
Keyword(s):  
Gene Expression ◽  
Kidney Function ◽  
Critical Role ◽  
Kidney Injury ◽  
Sequencing Analysis ◽  
Kidney Fibrosis ◽  
Atp Production ◽  
Expression Of Genes ◽  
Genes Encoding

Altered cellular metabolism in kidney proximal tubule (PT) cells plays a critical role in acute kidney injury (AKI). The transcription factor Krüppel-like factor 6 (KLF6) is rapidly and robustly induced early in the PT after AKI. We found that PT-specific Klf6 knockdown (Klf6PTKD) is protective against AKI and kidney fibrosis in mice. Combined RNA and chromatin immunoprecipitation sequencing analysis demonstrated that expression of genes encoding branched-chain amino acid (BCAA) catabolic enzymes was preserved in Klf6PTKD mice, with KLF6 occupying the promoter region of these genes. Conversely, inducible KLF6 overexpression suppressed expression of BCAA genes and exacerbated kidney injury and fibrosis in mice. In vitro, injured cells overexpressing KLF6 had similar decreases in BCAA catabolic gene expression and were less able to utilize BCAA. Furthermore, knockdown of BCKDHB, which encodes one subunit of the rate-limiting enzyme in BCAA catabolism, resulted in reduced ATP production, while treatment with BCAA catabolism enhancer BT2 increased metabolism. Analysis of kidney function, KLF6, and BCAA gene expression in human chronic kidney disease patients showed significant inverse correlations between KLF6 and both kidney function and BCAA expression. Thus, targeting KLF6-mediated suppression of BCAA catabolism may serve as a key therapeutic target in AKI and kidney fibrosis.

scholarly journals Obesity and Race Alter Gene Expression in Skin

2020 ◽  
Author(s):  
Jeanne M. Walker ◽  
Sandra Garcet ◽  
Jose O. Aleman ◽  
Christopher E. Mason ◽  
David Danko ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cystic Fibrosis ◽  
African Americans ◽  
Normal Weight ◽  
Gene Encoding ◽  
Functional Changes ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Expression Pathways ◽  
Altered Gene

ABSTRACTObesity is accompanied by dysfunction of many organs, but effects on the skin have received little attention. We studied differences in epithelial thickness by histology and gene expression by Affymetrix gene arrays and PCR in the skin of 10 obese (BMI 35-50) and 10 normal weight (BMI 18.5-26.9) postmenopausal women paired by age and race. Epidermal thickness did not differ with obesity but the expression of genes encoding proteins associated with skin blood supply and wound healing were altered. In the obese, many gene expression pathways were broadly downregulated and subdermal fat showed pronounced inflammation. There were no changes in skin microbiota or metabolites. African American subjects differed from Caucasians with a trend to increased epidermal thickening. In obese African Americans, compared to obese Caucasians, we observed altered gene expression that may explain known differences in water content and stress response. African Americans showed markedly lower expression of the gene encoding the cystic fibrosis transmembrane regulator characteristic of the disease cystic fibrosis. The results from this preliminary study may explain the functional changes found in the skin of obese subjects and African Americans.

scholarly journals Transcriptional Modulation of Genes Encoding Structural Characteristics of Differentiating Enterocytes During Development of a Polarized Epithelium In Vitro

2007 ◽  
Vol 18 (11) ◽  
pp. 4261-4278 ◽  
Author(s):  
Jennifer M. Halbleib ◽  
Annika M. Sääf ◽  
Patrick O. Brown ◽  
W. James Nelson
Keyword(s):  
Gene Expression ◽  
Cell Polarity ◽  
Structural Characteristics ◽  
Expression Patterns ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Functional Features ◽  
Cell Cell

Although there is considerable evidence implicating posttranslational mechanisms in the development of epithelial cell polarity, little is known about the patterns of gene expression and transcriptional regulation during this process. We characterized the temporal program of gene expression during cell–cell adhesion–initiated polarization of human Caco-2 cells in tissue culture, which develop structural and functional polarity similar to that of enterocytes in vivo. A distinctive switch in gene expression patterns occurred upon formation of cell–cell contacts between neighboring cells. Expression of genes involved in cell proliferation was down-regulated concomitant with induction of genes necessary for functional specialization of polarized epithelial cells. Transcriptional up-regulation of these latter genes correlated with formation of important structural and functional features in enterocyte differentiation and establishment of structural and functional cell polarity; components of the apical microvilli were induced as the brush border formed during polarization; as barrier function was established, expression of tight junction transmembrane proteins peaked; transcripts encoding components of the apical, but not the basal-lateral trafficking machinery were increased during polarization. Coordinated expression of genes encoding components of functional cell structures were often observed indicating temporal control of expression and assembly of multiprotein complexes.

scholarly journals Expression of genes encoding resistance in Staphylococcus spp. isolated from bovine subclinical mastitis in Brazil

2020 ◽  
Vol 14 (07) ◽  
pp. 772-780
Author(s):  
Eveline Zuniga ◽  
Nilson Roberti Benites ◽  
Aline Santana da Hora ◽  
Priscila Luiza Mello ◽  
Marco Antonio Laes ◽  
...  
Keyword(s):  
Methicillin Resistance ◽  
Meca Gene ◽  
Bovine Mastitis ◽  
Subclinical Mastitis ◽  
Gene Encoding ◽  
Resistance Factors ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Staphylococcus Spp

Introduction: Staphylococci are the most important agents associated with bovine mastitis. This study aimed at characterizing resistance factors to antimicrobials in Staphylococcus spp. isolated from the milk of cows with subclinical mastitis. Methodology: In vitro resistance of 243 Staphylococcus spp. isolates to antimicrobials commonly used in clinical practice was evaluated. The detection and expression of genes encoding resistance mecA (gene encoding penicillin binding protein 2a) mecALGA251 (mecA homologue), blaZ (gene encoding penicillin resistance), femA and femB (genes encoding essential factors - A and B - for the expression of methicillin resistance) and aacA-aphD (gene encoding for a bifunctional enzyme that confers resistance to gentamicin) using PCR and RT-PCR was investigated. Results: One or more genes encoding resistance to different antimicrobials were detected in 184 Staphylococcus spp. samples. The femA and femB genes were the most frequent. Regarding the variables’ detection (N = number of strains) and expression (% of strains), the following results were obtained: blaZ (N = 40 – 82.5%), femA (N = 147 – 47.6%), aacAaphD (N = 30 – 43.3%), femB (N = 138 – 29.7%), mecA (N = 33 – 27.3%), mecALGA251 (N = 01 – 0.0%). There was a higher occurrence of phenotypic resistant strains for amoxicillin, ampicillin and penicillin in isolates positive for detection and/or expression of blaZ gene when compared with the other genes. Conclusions: The present study provides new information on genotypic traits of Staphylococcus isolates from bovine subclinical mastitis especially regarding the evaluation of expression of genes associated with antimicrobial resistance in Staphylococcus spp. using molecular tools.

scholarly journals Activation of Transcription by Metabolic Intermediates of the Pyrimidine Biosynthetic Pathway

1999 ◽  
Vol 19 (1) ◽  
pp. 882-888 ◽  
Author(s):  
Paul J. Flynn ◽  
Richard J. Reece
Keyword(s):  
Gene Expression ◽  
Transcriptional Activation ◽  
Biosynthetic Pathway ◽  
De Novo ◽  
Activation Function ◽  
Expression Of Genes ◽  
Activation Of Transcription ◽  
Transcriptional Activator Protein ◽  
Genes Encoding

ABSTRACT Saccharomyces cerevisiae responds to pyrimidine starvation by increasing the expression of four URA genes, encoding the enzymes of de novo pyrimidine biosynthesis, three- to eightfold. The increase in gene expression is dependent on a transcriptional activator protein, Ppr1p. Here, we investigate the mechanism by which the transcriptional activity of Ppr1p responds to the level of pyrimidine biosynthetic intermediates. We find that purified Ppr1p is unable to promote activation of transcription in an in vitro system. Transcriptional activation by Ppr1p can be observed, however, if either dihydroorotic acid (DHO) or orotic acid (OA) is included in the transcription reactions. The transcriptional activation function and the DHO/OA-responsive element of Ppr1p localize to the carboxyl-terminal 134 amino acids of the protein. Thus, Ppr1p directly senses the level of early pyrimidine biosynthetic intermediates within the cell and activates the expression of genes encoding proteins required later in the pathway. These results are discussed in terms of (i) regulation of the pyrimidine biosynthetic pathway and (ii) a novel mechanism of regulating gene expression.

scholarly journals Expression of genes encoding zona pellucida glycoproteins and cortical granule distribution in porcine oocytes isolated from small and medium follicles in relation to puberty status of donors

2017 ◽  
Vol 62 (No. 6) ◽  
pp. 234-241 ◽  
Author(s):  
M. Jeseta ◽  
J. Budna ◽  
W. Kranc ◽  
S. Hanulakova ◽  
A. Bryja ◽  
...  
Keyword(s):  
Gene Expression ◽  
Zona Pellucida ◽  
Marker Gene ◽  
Cortical Granule ◽  
Cortical Granules ◽  
Expression Of Genes ◽  
Gamete Recognition ◽  
Genes Encoding ◽  
Central Concentration

The zona pellucida proteins belong to a group of proteins that regulate the processes of gamete recognition, interaction, and fusion, since they are recognized as primary and secondary sperm receptors. It is suggested that cortical granule distribution is significantly associated with the zona pellucida structure and membrane preparation to cortical and acrosome reaction. Therefore, this study investigated the zona pellucida marker gene expression (ZP2 and ZP4 protein) in relation to cortical granules distribution in oocytes and puberty status of donors. Oocytes were collected from adult cyclic sows (isolated from medium and small follicles) and juvenile gilts (isolated only from small follicles). The oocytes were examined by RT-qPCR and by confocal microscopy. The expression of genes for ZP2 and ZP4 protein in oocytes collected from small follicles from cycling sows was higher in comparison to oocytes from medium follicles or oocytes collected from small follicles from juvenile gilts (P < 0.001). We also observed increased expression of both ZP2 and ZP4 mRNA in oocytes collected from small follicles (juvenile gilts) as compared to medium follicles (cycling sows) (P < 0.001). Moreover, we found a difference in the distribution of cortical granules. In oocytes from medium follicles, the peripheral localization of cortical granules was twice higher than their central concentration. However, in oocytes derived from small follicles (cyclic sows) cortical granules in comparison to oocytes from medium follicles were more centrally localized (P < 0.05). It has been suggested that the donor puberty status and the size of follicles significantly influenced the zona pellucida gene expression and cortical granule localization in porcine oocytes. This is accompanied by fertilization specificity and fertilizability of porcine oocytes in vitro.

scholarly journals The Assessment of IL-21 and IL-22 at the mRNA Level in Tumor Tissue and Protein Concentration in Serum and Peritoneal Fluid in Patients with Ovarian Cancer

2021 ◽  
Vol 10 (14) ◽  
pp. 3058
Author(s):  
Aleksandra Mielczarek-Palacz ◽  
Celina Kruszniewska-Rajs ◽  
Marta Smycz-Kubańska ◽  
Jarosław Strzelczyk ◽  
Wojciech Szanecki ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Peritoneal Fluid ◽  
Tumor Tissue ◽  
Mrna Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
First Time ◽  
Ovarian Serous Cancer

The aim of the analysis was for the first time to assess the expression of genes encoding IL-21 and IL-22 at the mRNA level in ovarian tumor specimens and the concentration of these parameters in serum and peritoneal fluid in patients with ovarian serous cancer. The levels of IL-21 and IL-22 transcripts were evaluated with the use of the real-time RT-qPCR. Enzyme-linked immunosorbent assay (ELISA) was used to determine the concentration of proteins. Quantitative analysis of IL-21 gene mRNA in the tumor tissue showed the highest activity in the G1 degree of histopathological differentiation and was higher in G1 compared to the control group. The concentration of IL-21 and IL-22 in the serum and in the peritoneal fluid of women with ovarian cancer varied depending on the degree of histopathological differentiation of the cancer and showed statistical variability compared to controls. The conducted studies have shown that the local and systemic changes in the immune system involving IL-21 and IL-22 indicate the participation of these parameters in the pathogenesis of ovarian cancer, and modulation in the IL-21/IL-22 system may prove useful in the development of new diagnostic and therapeutic strategies used in patients, which require further research.

scholarly journals In vivo transcriptome analysis provides insights into host-dependent expression of virulence factors by Yersinia entomophaga MH96, during infection of Galleria mellonella

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Amber R Paulson ◽  
Maureen O’Callaghan ◽  
Xue-Xian Zhang ◽  
Paul B Rainey ◽  
Mark R H Hurst
Keyword(s):  
Galleria Mellonella ◽  
Functional Enrichment ◽  
Natural Environments ◽  
Insecticidal Toxin ◽  
Heat Stable ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Cell Densities

Abstract The function of microbes can be inferred from knowledge of genes specifically expressed in natural environments. Here, we report the in vivo transcriptome of the entomopathogenic bacterium Yersinia entomophaga MH96, captured during initial, septicemic, and pre-cadaveric stages of intrahemocoelic infection in Galleria mellonella. A total of 1285 genes were significantly upregulated by MH96 during infection; 829 genes responded to in vivo conditions during at least one stage of infection, 289 responded during two stages of infection, and 167 transcripts responded throughout all three stages of infection compared to in vitro conditions at equivalent cell densities. Genes upregulated during the earliest infection stage included components of the insecticidal toxin complex Yen-TC (chi1, chi2, and yenC1), genes for rearrangement hotspot element containing protein yenC3, cytolethal distending toxin cdtAB, and vegetative insecticidal toxin vip2. Genes more highly expressed throughout the infection cycle included the putative heat-stable enterotoxin yenT and three adhesins (usher-chaperone fimbria, filamentous hemagglutinin, and an AidA-like secreted adhesin). Clustering and functional enrichment of gene expression data also revealed expression of genes encoding type III and VI secretion system-associated effectors. Together these data provide insight into the pathobiology of MH96 and serve as an important resource supporting efforts to identify novel insecticidal agents.

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