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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

RNA polymerase II transcription is influenced both by how rapidly a gene is induced and by the rate at which continuous reinitiation occurs after induction. We show here that in vitro the rates of these two critical steps need not be the same. For activator GAL-AH-dependent HeLa transcription, the rate of assembling a preinitiation complex is significantly slower than the rate of reinitiation. Although reinitiation is rapid, it still requires ATP hydrolysis. This unexpected uncoupling of the rates of initiation and reinitiation implies that in regulating mammalian promoter activity, one must consider separately the controls on initiation during induction and the controls on the subsequent reinitiation events.
Mol Cell Biol 1993 Aug
PMID:Uncoupling of initiation and reinitiation rates during HeLa RNA polymerase II transcription in vitro. 833 2

In contrast to other members of the steroid/thyroid hormone superfamily, not much is known about the regions involved in transactivation of the receptors for retinoic acid. To determine the transactivation function of RARs, fusion proteins between the DNA-binding domain of the yeast transcription factor GAL4 and retinoic acid receptor-alpha (RAR alpha) or RAR beta were made. Transfection of these constructs resulted in RA-induced activation of a GAL4-responsive element-containing promoter. Deletion analysis revealed that RAR beta-2 has two transcription activation functions (TAFs). TAF-1 activates transcription constitutively and was mapped to the first 32 amino acids of the A-region. TAF-2 is located in the ligand-binding domain between amino acids 137 and 410 and activated transcription only in the presence of RA. The presence of two TAFs was confirmed by cotransfection of RAR beta deletion constructs with the human RAR beta-2 promoter as reporter, showing that the absence of RAR beta TAF-1 causes a decrease in transactivation, whereas truncation of TAF-2 completely blocks this function. Internal deletions in the ligand-binding domain in both GAL-RAR beta and RAR beta expression constructs resulted in a nonfunctional receptor, indicating that the complete ligand-binding domain is required for its transactivation function. Furthermore, we have shown that the contribution of the two TAFs in transcription activation varies among different cell lines, suggesting that they act in a cell-specific manner.
Mol Endocrinol 1993 Apr
PMID:The retinoic acid receptor-beta 2 contains two separate cell-specific transactivation domains, at the N-terminus and in the ligand-binding domain. 838 1

Glucose added to the medium was found to enhance superoxide production by isolated circulating neutrophils from both diabetic and normal subjects, but quantitatively the enhancement decreased from 4 to 50 mmole/liter. Galactose up to 50 mmole/liter had no effect on superoxide production in cells from the control subjects, but appeared to depress it in those from diabetics. No correlations were found between indices of the degree of hyperglycemia (plasma glucose and hemoglobin A1c) and the magnitude of the respiratory burst in cells from diabetics. When the isolated cells from normal and diabetic subjects were restored to a medium containing glucose at the original concentration in plasma at phlebotomy, the rate of superoxide production was approximately doubled in every case and there was no significant difference between diabetic and normal cells. Preincubation of cells for 1 hr in the presence of 0-50 mmole/liter glucose or galactose prior to activation had no significantly depressant effect on the respiratory burst except at 50 mmole/liter glucose in diabetic cells. It is concluded that circulating neutrophils from the diabetic population under the conditions studied are just as competent as control cells in their ability to sustain superoxide production over a wide range of energy availability.
Exp Mol Pathol 1993 Jun
PMID:Superoxide production by neutrophils from diabetics and normal subjects in response to glucose and galactose. 839 Sep 41

The GAL4 gene of Saccharomyces cerevisiae (encoding the activator of transcription of the GAL genes) is poorly expressed and is repressed during growth on glucose. To determine the basis for its weak expression and to identify DNA sequences recognized by proteins that activate transcription of a gene that itself encodes an activator of transcription, we have analyzed GAL4 promoter structure. We show that the GAL4 promoter is about 90-fold weaker than the strong GAL1 promoter and at least 7-fold weaker than the feeble URA3 promoter and that this low level of GAL4 expression is primarily due to a weak promoter. By deletion mapping, the GAL4 promoter can be divided into three functional regions. Two of these regions contain positive elements; a distal region termed the UASGAL4 (upstream activation sequence) contains redundant elements that increase promoter function, and a central region termed the UESGAL4 (upstream essential sequence) is essential for even basal levels of GAL4 expression. The third element, an upstream repression sequence, mediates glucose repression of GAL4 expression and is located between the UES and the transcriptional start site. The UASGAL4 is unusual because it is not interchangable with UAS elements in other yeast promoters; it does not function as a UAS element when inserted in a CYC1 promoter, and a normally strong UAS functions poorly in place of UASGAL4 in the GAL4 promoter. Similarly, the UES element of GAL4 does not function as a TATA element in a test promoter, and consensus TATA elements do not function in place of UES elements in the GAL4 promoter. These results suggest that GAL4 contains a weak TATA-less promoter and that the proteins regulating expression of this regulatory gene may be novel and context specific.
Mol Cell Biol 1993 Aug
PMID:Promoter elements determining weak expression of the GAL4 regulatory gene of Saccharomyces cerevisiae. 839 42

A new modular gene-expression system for use in studies of translational control in Saccharomyces cerevisiae was constructed. A GAL::PGK fusion promoter (GPF) directed the inducible synthesis of mRNAs initiated at a single major site. A series of leader sequences were tested in combination with each of two reporter genes (encoding chloramphenicol acetyl transferase (cat) and luciferase (luc)). Stem-loop structures of three different sizes and predicted stabilities were inserted into each of two different unique restriction sites in the leader. After correction for relative mRNA abundance, a stem-loop of predicted stability equivalent to approximately -18 kcal mol-1 inhibited translation by up to 89%. The degree of inhibition exerted by the other stem-loops correlated positively with their predicted stabilities. Combinations of two stem-loops at different sites yielded an inhibitory effect greater than that of either individual stem-loop alone. Similar inhibitory effects were observed with both reporter genes. However, inhibition of translation, particularly of the cat gene, was more effective when the stem-loop was positioned close to the start codon rather than at the 5' end of the leader. The observed results reflect an important form of post-transcriptional control that is expected to act on a large number of genes in yeast.
Mol Microbiol 1993 Aug
PMID:Inhibition of translational initiation in Saccharomyces cerevisiae by secondary structure: the roles of the stability and position of stem-loops in the mRNA leader. 841 99

The concentration of the transcriptional activator LAC9 (KlGAL4) of Kluyveromyces lactis is moderately regulated by the carbon source as is the case for GAL4, its homolog in Saccharomyces cerevisiae. Expression of the LAC9 gene is induced about twofold in galactose. This induction is due to autoregulation. The LAC9 gene product binds to a low-affinity binding site in the LAC9 promoter and moderately activates transcription in response to galactose above a basal level. As for the LAC9-controlled metabolic genes, induction of LAC9 is inhibited in the presence of glucose. This inhibition of induction is a prerequisite for glucose repression of the lactose-galactose metabolic pathway. On the other hand, induced LAC9 levels are required for optimal growth on galactose, since mutating the LAC9 binding site in the LAC9 promoter resulted in poor growth and reduced expression of LAC9-controlled genes. Thus, in addition to the GAL80-dependent regulation by protein-protein interaction, the regulation of LAC9 gene expression is an important parameter in determining carbon source control of the LAC-GAL regulon. Although the mode of control is different, the pattern of LAC9 gene regulation resembles that of the S. cerevisiae GAL4 gene, being lower in glucose and glucose-galactose than in galactose.
Mol Cell Biol 1993 May
PMID:Expression of the transcriptional activator LAC9 (KlGAL4) in Kluyveromyces lactis is controlled by autoregulation. 847 61

Two Saccharomyces cerevisiae proteins of 21 and 27 kDa co-purify with a novel enhancer of Gal4p DNA binding activity (Egdp) [Parthun et al., Mol. Cell. Biol. 12 (1992) 5683-5689]. Mutations in the EGD1 gene encoding the 21-kDa protein (Egd1p) have been shown to affect the kinetics and extent of the Gal4p-mediated, galactose-induced activation of the GAL genes. Egd1p is homologous to human BTF3b, recently identified as the beta subunit of the heterodimeric nascent-polypeptide-associated complex (NAC) involved in ensuring signal-sequence-specific protein sorting and translocation [Wiedmann et al., Nature 370 (1994) 434-440]. We have cloned and characterized EGD2 encoding the 27-kDa protein and found that Egd2p is strikingly similar to the alpha subunit of human NAC. Yeast, therefore, contains a complex composed of Egd1p and Egd2p very similar to the NAC complex described in human cells. Disruption of EGD2, alone or in combination with an EGD1 disruption, causes no obvious phenotypes. The lack of phenotype, the high levels of EGD1 and EGD2 expression, and the identification of multiple human genes encoding NAC subunits suggest that the yeast EGD genes may be members of multigene families with redundant function.
 ...
PMID:The yeast EGD2 gene encodes a homologue of the alpha NAC subunit of the human nascent-polypeptide-associated complex. 852 75

We describe the expression of gpIRK1, an inwardly rectifying K+ channel obtained from guinea pig cardiac cDNA. gpIRK1 is a homologue of the mouse IRK1 channel identified in macrophage cells. Expression of gpIRK1 in Xenopus oocytes produces inwardly rectifying K+ current, similar to the cardiac inward rectifier current IK1. This current is blocked by external Ba2+ and Cs+. Plasmids containing the gpIRK1 coding region under the transcriptional control of constitutive (PGK) or inducible (GAL) promoters were constructed for expression in Saccharomyces cerevisiae. Several observations suggest that gpIRK1 forms functional ion channels when expressed in yeast. gpIRK1 complements a trk1 delta trk2 delta strain, which is defective in potassium uptake. Expression of gpIRK1 in this mutant restores growth on low potassium media. Growth dependent on gpIRK1 is inhibited by external Cs+. The strain expressing gpIRK1 provides a versatile genetic system for studying the assembly and composition of inwardly rectifying K+ channels.
Mol Biol Cell 1995 Sep
PMID:Functional expression of a vertebrate inwardly rectifying K+ channel in yeast. 853 18

Galactose metabolism in all organisms is catalyzed by three enzymatic steps: the galactokinase, galactose-1-phosphate uridyltransferase, and UDP galactose 4'-epimerase reactions. We report here the molecular cloning, characterization, and mapping of a full-length cDNA encoding human UDP-galactose 4'-epimerase (GALE). Our cDNA is 1488 bp long and matches the mRNA size of 1.5 kg detected in fibroblasts and lymphoblasts. The human GALE cDNA encodes a predicted protein of 348 amino acids with a molecular mass of 38,266. The human GALE enzyme is 87% identical to the rat protein, 53% identical to the homologous GAL10 protein from the yeast Kluyveromyces lactis, and 51% identical to the galE protein from the prokaryote Escherichia coli. This extraordinary degree of sequence identity has allowed us to build a homology model of the human protein based on the bacterial crystal structure. This predicted human structure is very similar to the E. coli galE enzyme, suggesting that both enzymes use similar mechanisms. The human gene encoding GALE maps, as expected, to a single locus on chromosome 1 and appears to be compact. The human GALE gene is structurally intact in 19 patients with epimerase-deficiency galactosemia, an inborn error of metabolism secondary to GALE deficiency. Therefore, we propose that this disorder is due to small mutations within the gene.
Biochem Mol Med 1995 Oct
PMID:Molecular cloning, characterization, and mapping of a full-length cDNA encoding human UDP-galactose 4'-epimerase. 859 31

Transcriptional activation is thought to be mediated by DNA-bound activators through interaction with a basal transcription factor thereby stabilizing the pre-initiation complex. For such interaction cofactors such as TAFs, bridging proteins, mediators or intermediary proteins are required by binding simultaneously to the activator and the target. We have investigated the activation functions (AFs) of both RARbeta and RXRalpha and show that both activators contain two homologous AFs. By comparing the capacity to activate transcription by these AFs on several promoters, both as full-length receptors and as fusion-proteins of AFs with the DNA-binding domain of the yeast transcription factor GAL-4, we were able to show that these AFs function by different mechanisms. We found that the activity of these AFs is cell-type specific, as they are more active in certain cell lines than in others. Furthermore we observed that the AFs of RARbeta and RXRalpha can activate transcription synergistically both as GAL-fusion protein and with full-length receptors. For AF-2 of RAR beta we observed cell type-dependent difference in synergistic activation and we show that the E1A protein, which functions as a cofactor for RAR beta, permits synergistic activation in cell lines in which in the absence of this cofactor transcription occurs non-synergistically. We propose a model in which several non cell type specific cofactors and cell-specific cofactors act together to form a more stable pre-initiation complex explaining the observed cell-specific synergistic activation.
J Steroid Biochem Mol Biol 1996 Jan
PMID:A role for cofactors in synergistic and cell-specific activation by retinoic acid receptors and retinoid X receptor. 860 32


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