Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

CrgA is a LysR-type transcriptional regulator involved in the intimate adhesion of Neisseria meningitidis to target human epithelial cells. It is negatively autoregulated, and its expression is transiently induced upon contact with target cells. We analysed the functional organization of CrgA using in frame deleted proteins. Four truncated proteins were constructed and purified. They were deleted between residues 20 and 40, 121 and 154, 111 and 181 and 268 and 291. Meningococcal mutants harbouring the corresponding deleted crgA alleles were also constructed. All mutants showed a reduced ability to adhere to epithelial cells. beta-Galactosidase activity assays using a crgA-lacZ transcriptional fusion showed that all the mutations except the 268-291 deletion resulted in loss of induction upon contact with target cells. Gel mobility shift assays and cross-linking assays showed that the oligomerization of CrgA is required for DNA binding and that the N-terminal part of CrgA is directly involved in DNA binding through a helix-turn-helix motif. The C-terminal region is also involved in DNA binding, probably by permitting the oligomerization of CrgA. The C-terminal region also seemed to interact with RNA polymerase. Therefore, the binding of CrgA and its interaction with RNA polymerase may inhibit the clearance of meningococcal promoters that are repressed by CrgA during the intimate adhesion of N. meningitidis to target cells.
Mol Microbiol 2003 Jan
PMID:The Neisseria meningitidis adhesion regulatory protein CrgA acts through oligomerization and interaction with RNA polymerase. 1249 59

We examined regulation of the Na+/H+ exchanger (NHE1 isoform) in the developing mouse. We generated transgenic mice with the Na+/H+ exchanger promoter directing expression of the beta-Galactosidase reporter. We found that expression of the Na+/H+ exchanger was maximum in the heart and liver of 12-day-old embryonic mice. Similar results were found in mice using the green fluorescent protein reporter driven by the Na+/H+ exchanger promoter. Detailed examination of the myocardium revealed that the GFP reporter protein was expressed in the cytoplasm of cardiomyocyte cells. We examined NHE1 protein expression in transgenic mice lacking the transcription factors AP-2alpha or the transcription factor COUP-TF1. Eighteen-day-old AP-2alpha heterozygote mice show no large changes in NHE1 expression in heart, lung, liver, kidney and brain. In contrast, 18-day-old embryos from AP-2alpha null mice showed a large increase in Na+/H+ exchanger protein expression in the brain. NHE1 protein levels in COUP-TF1 knockout embryos did not differ from wild type embryos. The results suggest that AP-2alpha and COUP-TF1 are not critical to NHE1 expression in the late stage embryo and that other related transcription factors may function in regulation of the Na+/H+ exchanger.
Mol Cell Biochem 2003 Jan
PMID:Transcriptional regulation of Na+/H+ exchanger expression in the intact mouse. 1261 93

A cluster of genes coding for putative plant cell-wall degrading enzymes (i.e., genes for two endoglucanases [gunA and gunA2], one pectinmethylesterase [pme], and one polygalacturonase [pgl]) was identified by sequence similarities in the symbiotic region of the Bradyrhizobium japonicum chromosome. In addition, a systematic screen of the region revealed several genes potentially transcribed by the sigma(54)-RNA polymerase and activated by the transcriptional regulator NifA (i.e., genes for proteins with similarity to outer membrane proteins [id117 and id525] and a citrate carrier [id331 or citA] and one open reading frame without similarity to known proteins [id747]). Expression studies using transcriptional lacZ fusions showed that gunA2 and pgl were strongly induced by the isoflavone genistein in a NodW-dependent manner, suggesting a role of the gene products in early events of the nodulation process; by contrast, gunA and pme expression was very weak in the conditions tested. The gunA2 gene product was purified and was shown to have cellulase activity. beta-Galactosidase activity expressed from transcriptional lacZ fusions to id117, id525, and id747 in the wild type and in nifA and rpoN mutant backgrounds confirmed that their transcription was dependent on NifA and sigma(54). Despite the presence of a -24/-12-type promoter and a NifA binding site upstream of citA, no regulation could be demonstrated in this case. Null mutations introduced in gunA, gunA2, pgl, pme, citA, id117, id525, and id747 did not impair the symbiosis with the host plants.
Mol Plant Microbe Interact 2003 Apr
PMID:New NodW- or NifA-regulated Bradyrhizobium japonicum genes. 1274 63

Biosynthetic pathways for heme and chlorophyll share common intermediates from 5-aminolevulinic acid through protoporphyrin IX. To obtain a better understanding of how photosynthetic organisms coordinate heme and chlorophyll biosynthesis, we have undertaken detailed analysis of the expression pattern of numerous heme biosynthesis genes in the purple photosynthetic bacterium Rhodobacter capsulatus. beta-Galactosidase reporter assays demonstrated that expression of hemA, hemB, hemC, hemE and hemZ genes is elevated under conditions that give rise to elevated bacteriochlorophyll synthesis. Heme gene expression is shown to be affected by mutations in previously identified transcriptional regulators RegA, FnrL, CrtJ, and AerR, which also control expression of genes involved in bacteriochlorophyll and carotenoid synthesis, and synthesis of the apoprotein subunits of the photosynthetic and electron transport apparatus. High-resolution primer extension analysis of hem mRNA reveals the presence of numerous putative RegA, FnrL and CrtJ binding sites in several hem promoter regions.
J Mol Biol 2004 Sep 24
PMID:Regulation of hem gene expression in Rhodobacter capsulatus by redox and photosystem regulators RegA, CrtJ, FnrL, and AerR. 1535 43

This chapter provides information on beta-galactosidase staining of whole mouse embryos, organs, tissue sections, and cultured cells, as well as double staining with horseradish peroxidase and use as a tool for genotyping. Using these protocols, localization of beta-galactosidase can be visualized throughout development and in adult tissues. beta-Galactosidase staining may be used purely as a marker of gene expression and also as a tracer in cell lineage studies.
Methods Mol Biol 2007
PMID:Methodologies for staining and visualisation of beta-galactosidase in mouse embryos and tissues. 1828 34

G(M1) gangliosidosis is an inherited, fatal neurodegenerative disease caused by deficiency of lysosomal beta-d-galactosidase (EC 3.2.1.23) and consequent storage of undegraded G(M1) ganglioside. To characterize the genetic mutation responsible for feline G(M1) gangliosidosis, the normal sequence of feline beta-galactosidase cDNA first was defined. The feline beta-galactosidase open reading frame is 2010 base pairs, producing a protein of 669 amino acids. The putative signal sequence consists of amino acids 1-24 of the beta-galactosidase precursor protein, which contains seven potential N-linked glycosylation sites, as in the human protein. Overall sequence homology between feline and human beta-galactosidase is 74% for the open reading frame and 82% for the amino acid sequence. After normal beta-galactosidase was sequenced, the mutation responsible for feline G(M1) gangliosidosis was defined as a G to C substitution at position 1448 of the open reading frame, resulting in an amino acid substitution at arginine 483, known to cause G(M1) gangliosidosis in humans. Feline beta-galactosidase messenger RNA levels were normal in cerebral cortex, as determined by quantitative RT-PCR assays. Although enzymatic activity is severely reduced by the mutation, a full-length feline beta-galactosidase cDNA restored activity in transfected G(M1) fibroblasts to 18-times normal. beta-Galactosidase protein levels in G(M1) tissues were normal on Western blots, but immunofluorescence analysis demonstrated that the majority of mutant beta-galactosidase protein did not reach the lysosome. Additionally, G(M1) cat fibroblasts demonstrated increased expression of glucose-related protein 78/BiP and protein disulfide isomerase, suggesting that the unfolded protein response plays a role in pathogenesis of feline G(M1) gangliosidosis.
Mol Genet Metab 2008 Jun
PMID:Molecular consequences of the pathogenic mutation in feline GM1 gangliosidosis. 1835 97

A generic whole-cell bacterial sensor called sposensor was developed with immobilized spores from engineered Bacillus subtilis. Sposensor contains two different types of spores: reporting spores that contain a reporter gene fused to a promoter responding to a compound to be detected, and control spores use to monitor cell germination and viability. A one-step incubation/detection process was developed to meet the constraints of on-site analysis. Spores were directly incubated with culture medium containing the compound to be detected. beta-Galactosidase was chosen as a reporter protein in both cases and its activity followed by a colorimetric assay. Results showed that sposensor was efficient in detecting two different compounds, a metal (Zn(2+)) and a peptidic antibiotic (bacitracin). Owing to the stability and robustness of spores, sposensor is a very efficient and easy tool to manipulate for analyzing the presence of toxic compounds in natural settings.
J Mol Microbiol Biotechnol 2009
PMID:Sposensor: a whole-bacterial biosensor that uses immobilized Bacillus subtilis spores and a one-step incubation/detection process. 1925 7

We describe a fast, simple assay for testing ligands for binding to an unknown receptor after transfection of that receptor cDNA into tissue culture cells. The assay is based on a mouse L cell line (LVIP2.OZc) that contains a cyclic AMP responsive reporter construct and is performed in 96-well plates. If the appropriate agonist binds to a receptor clone coupled to G(s) proteins, activation of adenylyl cyclase produces cAMP which in turn induces the enzyme beta-galactosidase in LVIP2.0Zc cells. beta-Galactosidase activity is detected by staining cells with a chromogenic substrate. After cell lysis, incubation with o-nitrophenyl beta-d-galactopyranoside (ONPG) results in a yellow color. Color development can be observed with the naked eye or read with a plate reader at 405 nm. Agonists that bind to G(i)-coupled receptors can be identified by inhibition of forskolin-induced expression of beta-galactosidase.
Mol Cell Neurosci 1991 Aug
PMID:Method for identifying ligands that bind to cloned G(s)- or G(i)-coupled receptors. 1991 16


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