scholarly journals DGCR14 InducesIl17aGene Expression through the RORγ/BAZ1B/RSKS2 Complex

2014 ◽  
Vol 35 (2) ◽  
pp. 344-355 ◽  
Author(s):  
Ichiro Takada
Keyword(s):  
Cell Differentiation ◽  
Mrna Expression ◽  
Digeorge Syndrome ◽  
Heart Defects ◽  
Biochemical Purification ◽  
S6 Kinase ◽  
Ionization Time ◽  
Flight Mass Spectrometry ◽  
Ribosomal S6 Kinase

TheDgcr14/Es2gene is located in a chromosomal region the loss of which has been associated with DiGeorge syndrome, a cause of immunodeficiency, heart defects, and skeletal abnormalities. However, the role of DGCR14 protein remains to be elucidated. Here, I found that DGCR14 protein acts as a coactivator of RORγt in TH17 cells. Biochemical purification of the RORγ coregulator complex allowed me to identify the associated DGCR14 protein by matrix-assisted laser desorption ionization–time of flight mass spectrometry. Overexpression ofDgcr14mRNA enhanced RORγt-mediated transcriptional activity and facilitated TH17 cell differentiation. Furthermore, knockdown of Dgcr14 reducedIl17amRNA expression. I also found that DGCR14 associated with ribosomal S6 kinase 2 (RSK2, also called RpS6ka3) and BAZ1B, both of which were recruited to theIl17apromoter during TH17 cell differentiation. Knockdown ofBaz1borRpS6ka3also reducedIl17amRNA expression, andBaz1bknockdown increased transcriptional suppressive histone marks (histone H3K9me3) on theIl17apromoter. My findings showed the roles of DGCR14, RSK2, and BAZ1B in the transcriptional regulation ofIl17amRNA during TH17 cell differentiation.

scholarly journals ScaC, an Adaptor Protein Carrying a Novel Cohesin That Expands the Dockerin-Binding Repertoire of the Ruminococcus flavefaciens 17 Cellulosome

2004 ◽  
Vol 186 (9) ◽  
pp. 2576-2585 ◽  
Author(s):  
Marco T. Rincón ◽  
Jennifer C. Martin ◽  
Vincenzo Aurilia ◽  
Sheila I. McCrae ◽  
Garry J. Rucklidge ◽  
...  
Keyword(s):  
Adaptor Protein ◽  
Structural Components ◽  
Ruminococcus Flavefaciens ◽  
Ionization Time ◽  
One Dimensional ◽  
Native Proteins ◽  
Flight Mass Spectrometry ◽  
C Terminus ◽  
New Gene

ABSTRACT A new gene, designated scaC and encoding a protein carrying a single cohesin, was identified in the cellulolytic rumen anaerobe Ruminococcus flavefaciens 17 as part of a gene cluster that also codes for the cellulosome structural components ScaA and ScaB. Phylogenetic analysis showed that the sequence of the ScaC cohesin is distinct from the sequences of other cohesins, including the sequences of R. flavefaciens ScaA and ScaB. The scaC gene product also includes at its C terminus a dockerin module that closely resembles those found in R. flavefaciens enzymes that bind to the cohesins of the primary ScaA scaffoldin. The putative cohesin domain and the C-terminal dockerin module were cloned and overexpressed in Escherichia coli as His6-tagged products (ScaC-Coh and ScaC-Doc, respectively). Affinity probing of protein extracts of R. flavefaciens 17 separated in one-dimensional and two-dimensional gels with recombinant cohesins from ScaC and ScaA revealed that two distinct subsets of native proteins interact with ScaC-Coh and ScaA-Coh. Furthermore, ScaC-Coh failed to interact with the recombinant dockerin module from the enzyme EndB that is recognized by ScaA cohesins. On the other hand, ScaC-Doc was shown to interact specifically with the recombinant cohesin domain from ScaA, and the ScaA-Coh probe was shown to interact with a native 29-kDa protein spot identified as ScaC by matrix-assisted laser desorption ionization—time of flight mass spectrometry. These results suggest that ScaC plays the role of an adaptor scaffoldin that is bound to ScaA via the ScaC dockerin module, which, via the distinctive ScaC cohesin, expands the range of proteins that can bind to the ScaA-based enzyme complex.

Role of TPO in the Differentiation of Embryonic Stem Cells into Hematopoetic Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4202-4202
Author(s):  
Zheng Wang ◽  
Pramono Andri ◽  
Skokowa Julia ◽  
Welte Karl
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Rhesus Monkey ◽  
Embryonic Stem Cell ◽  
Mrna Expression ◽  
Es Cells ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Embryonic Stem Cell Differentiation

Abstract Thrombopoetin (TPO) is a primary regulator of megakaryocyte and platelet production. However, studies in c-mpl-deficient mice and in congenital amegakaryocytic thrombocytopenia-patients with non-sense c-mpl mutation who develop pancytopenia during the first years of life suggest that TPO also play an important role on early hematopoesis. We demonstrated that TPO enhances FLK-1 (VEGF-receptor) expression on hemangioblasts during murine embryonic stem cell differentiation in embryoid body-liquid cultures (up to 73%). To extend our studies, we investigated the TPO signaling in FLK-1 positive cells. ES cells at different time point of differentiation showed that TPO enhances c-mpl-, BMP4-, Notch-, HOXB4-, HOXB9-, HOXA10-, Runx1-and CD133- mRNA expression. To investigate mesoderm formation, we also analyzed GATA-4 and T-brachyury mRNA level expression. Interestingly, we found that TPO alone did not increase GATA-4- and T-brachyury- mRNA expression, suggesting that TPO requires other cytokines to form the mesoderm. We also found that TPO could maintain VEGF-A mRNA expression level during differentiation of ES-cells. We hypothesize that VEGF expression together with c-mpl expression is required in hematopoetic differentiation of ES cell. This activity of Tpo was also observed during Rhesus monkey embryonic stem cell differentiation into hematopoetic cell. Only combinations of TPO and VEGF were capable of increasing CD34 positive hematopoietic progenitor cells (up to 8%), but TPO alone failed to induce high levels of CD34+ cell. In addition, analysis of gene expression during hemangioblast development demonstrated that TPO was capable of increasing the expression of VEGF receptors (FLK-1) and TPO receptors (c-mpl) in mice and primates. The in-vitro differentiation of mouse and rhesus monkey ES cells provides an opportunity to better understand the role of TPO in the early stage of hematopoietic development from ES cells to mature hematopoietic cells.

scholarly journals Pathogenicity Factors and Antibiotic Resistance of the Bacteroides Fragilis

2020 ◽  
Author(s):  
S.S. Kozhakhmetova ◽  
E.V. Zholdybayeva ◽  
K.E. Mukhtarova ◽  
Ye.M. Ramankulov
Keyword(s):  
Antibiotic Resistance ◽  
Genomic Structure ◽  
Spectrometry Method ◽  
Ionization Time ◽  
Pathogenicity Factors ◽  
Flight Mass Spectrometry ◽  
Carbapenem Resistant ◽  
Genome Wide ◽  
Mechanisms Of Development

This article presents novel ideas about classification, genomic structure (inverted regions, mobile genetic elements, plasmids, mobilized and conjugated transposons), pathogenicity factors (adhesins, various enzymes, toxins, in particular, data on enterotoxin fragmentinis BFT - B. fragilis toxin), and the role of their metabolites in the manifestation of pathogenicity. Data on the global prevalence of antibiotic resistance in the clinical B. fragilis strains are presented. Mechanisms of development of the drug resistance are considered and the role of cfiA, tet, nim genes in the development of antibiotic resistance is disclosed. Information on the use of the MALDI-TOF MS (matrix-activated laser desorption-ionization time-of-flight mass spectrometry) method for distinguishing B.fragilis strains into two groups based on the ability to carry carbapenem resistant gene (carrying and not carrying cfiA gene) are presented. Basics of modes of emergence of multi-resistance in clinical strains of B. fragilis are considered. In addition, prospects for genome-wide sequencing in predicting antimicrobial resistance are presented. Currently increasing attention of researchers is payed to increase in resistance of B. fragilis to widely used antimicrobials. This is indeed of a great importance when choosing adequate antimicrobial therapy.

scholarly journals Implementation of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry in Routine Clinical Laboratories Improves Identification of Coagulase-Negative Staphylococci and Reveals the Pathogenic Role of Staphylococcus lugdunensis

2015 ◽  
Vol 53 (7) ◽  
pp. 2030-2036 ◽  
Author(s):  
Xavier Argemi ◽  
Philippe Riegel ◽  
Thierry Lavigne ◽  
Nicolas Lefebvre ◽  
Nicolas Grandpré ◽  
...  
Keyword(s):  
Laser Desorption Ionization ◽  
Coagulase Negative Staphylococci ◽  
Ionization Time ◽  
Content Type ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Phenotypic Methods ◽  
Tof Ms

The use of matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) for staphylococcal identification is now considered routine in laboratories compared with the conventional phenotypical methods previously used. We verified its microbiological relevance for identifying the main species of coagulase-negative staphylococci (CoNS) by randomly selecting 50 isolates. From 1 January 2007 to 31 August 2008, 12,479 staphylococci were isolated with phenotypic methods, of which 4,594 were identified asStaphylococcus aureusand 7,885 were coagulase negative staphylococci. Using MALDI-TOF MS from 1 January 2011 to 31 August 2012, 14,913 staphylococci were identified, with 5,066 asS. aureusand 9,847 as CoNS. MALDI-TOF MS allowed the identification of approximately 85% of the CoNS strains, whereas only 14% of the CoNS strains were identified to the species level with phenotypic methods because they were often considered contaminants. Furthermore, the use of MALDI-TOF MS revealed the occurrence of recently characterizedStaphylococcusspecies, such asS. pettenkoferi,S. condimenti, andS. piscifermentans. Microbiological relevance analysis further revealed that some species displayed a high rate of microbiological significance, i.e., 40% of theS. lugdunensisstrains included in the analysis were associated with infection risk. This retrospective microbiological study confirms the role of MALDI-TOF MS in clinical settings for the identification of staphylococci with clinical consequences. The species distribution reveals the occurrence of the recently identified speciesS. pettenkoferiand putative virulent species, includingS. lugdunensis.

scholarly journals Role of p90 Ribosomal S6 Kinase–Mediated Prorenin-Converting Enzyme in Ischemic and Diabetic Myocardium

Circulation ◽  
2006 ◽  
Vol 113 (14) ◽  
pp. 1787-1798 ◽  
Author(s):  
Seigo Itoh ◽  
Bo Ding ◽  
Tetsuro Shishido ◽  
Nicole Lerner-Marmarosh ◽  
Nadan Wang ◽  
...  
Keyword(s):  
Converting Enzyme ◽  
S6 Kinase ◽  
Diabetic Myocardium ◽  
P90 Ribosomal S6 Kinase ◽  
Ribosomal S6 Kinase

scholarly journals Role of p90 ribosomal S6 kinase in long-term synaptic facilitation and enhanced neuronal excitability

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Rong-Yu Liu ◽  
Yili Zhang ◽  
Paul Smolen ◽  
Leonard J. Cleary ◽  
John H. Byrne
Keyword(s):  
Neuronal Excitability ◽  
S6 Kinase ◽  
Synaptic Facilitation ◽  
P90 Ribosomal S6 Kinase ◽  
Ribosomal S6 Kinase
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