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D S Latchman

Publications and source records attributed to D S Latchman.

At least 145 records · Page 8Linked to original sources
PubMed1998

The TAATGARAT motif in the herpes simplex virus immediate-early gene promoters can confer both positive and negative responses to cellular octamer-binding proteins when it is located within the viral genome.

The TAATGARAT motif in the herpes simplex virus (HSV) immediate-early (IE) gene promoters plays a key role in their activation by the Oct-1-Vmw65 complex, but its role in mediating inhibitory effects of cellular octamer-binding proteins is less clear. We have used indicator viruses containing reporter constructs with different IE promoters driving a reporter beta-galactosidase gene within the viral genome to investigate this. We showed that deletion of the upstream IE promoter region containing the TAATGARAT motifs abolishes the inhibitory effect of the cellular octamer-binding proteins Oct-2.4 and Oct-2.5 on the viral IE promoter. This inhibitory effect can be restored by addition of a single TAATGARAT motif to the minimal promoter within the viral genome. Hence, the TAATGARAT motif can indeed mediate both positive and negative effects of cellular transcription factors when it is located within the viral genome.

Animals↗
PubMed1998

POU transcription factors Brn-3a and Brn-3b interact with the estrogen receptor and differentially regulate transcriptional activity via an estrogen response element.

The estrogen receptor (ER) modulates transcription by forming complexes with other proteins and then binding to the estrogen response element (ERE). We have identified a novel interaction of this receptor with the POU transcription factors Brn-3a and Brn-3b which was independent of ligand binding. By pull-down assays and the yeast two-hybrid system, the POU domain of Brn-3a and Brn-3b was shown to interact with the DNA-binding domain of the ER. Brn-3-ER interactions also affect transcriptional activity of an ERE-containing promoter, such that in estradiol-stimulated cells, Brn-3b strongly activated the promoter via the ERE, while Brn-3a had a mild inhibitory effect. The POU domain of Brn-3b which interacts with the ER was sufficient to confer this activation potential, and the change of a single amino acid in the first helix of the POU homeodomain of Brn-3a to its equivalent in Brn-3b can change the mild repressive effect of Brn-3a to a stimulatory Brn-3b-like effect. These observations and their implications for transcriptional regulation by the ER are discussed.

Binding Sites↗
PubMed1998

Analysis of the association of a heat shock protein70-1 gene promoter polymorphism with myocardial infarction and coronary risk traits.

Heat shock proteins (HSP) are induced during coronary ischaemia, and abnormal expression of one HSP gene may cause hypertension in rats. We examined association of a promoter polymorphism in the major stress-inducible hsp70 gene (hsp70-1 or HSP70A1) on chromosome 6 (p21.3) with coronary disease traits. This C-->A base substitution (AAACCCC) is at nucleotide position-110 in the heat shock transcription factor binding site (heat shock element, HSE). The first study sample (ECTIM), recruited from Belfast and three centers in France, consisted of 578 myocardial infarction cases and 698 age-matched controls. The frequency of the A-110 allele was 0.381 (95% CI = 0.35-0.41) and 0.384 (95% CI = 0.36-0.41) in cases and controls respectively. Homozygotes for the rarer A-110 allele had a higher BMI (27.3 kg/m2 +/- 3.9) compared with homozygotes for the common C-110 allele (26.3 kg/m2 +/- 3.3). The rarer homozygotes were shorter and heavier than the common homozygotes. A follow-up study involved 1431 healthy, middle aged men from the UK (NPHS II group). The frequency of the A-110 allele was 0.385 (95% CI = 0.37-0.40), and there was no association of genotype with BMI. Thus there appears to be no strong association of the Hsp70-1 promoter polymorphism with risk of myocardial infarction, BMI or any coronary disease traits analysed here.

Adult↗
PubMed1998

Heat shock proteins: protective effect and potential therapeutic use (review).

The heat shock proteins (hsps) are induced by a variety of stressful stimuli and their overexpression has been shown to protect cells both in vitro and in vivo against such stimuli, as well as against stimuli-inducing apoptosis. The potential therapeutic benefit of elevating hsp levels in individuals with, for example, cerebral or cardiac ischaemia or neurodegenerative diseases has led to the identification of specific methods of inducing hsp expression in a non-stressful manner. These include pharmacological procedures and cytokine treatment to elevate endogenous hsp levels and the development of viral vectors to deliver exogenous hsp genes. The advantages and disadvantages of each of these methods and their ultimate therapeutic potential are discussed.

Apoptosis↗
PubMed1997

C/EBP activates the human corticotropin-releasing hormone gene promoter.

The purpose of these studies was to identify whether transcription factors, associated with cytokine signalling, affected promoter activity of the corticotropin releasing hormone (CRH) gene. Fragments of a 3.6 kb sequence of the human CRH gene promoter were amplified by PCR and ligated upstream of a CAT reporter. These constructs were transfected into a variety of cell lines, either alone or together, with transcription factor expression vectors. Basal activity of a 3070 bp CRH promoter fragment was only seen in neuronal and lymphoblastoid cell lines. Promoter activity was increased by the transcription factors C/EBPbeta (NF-IL6) and more strongly, by C/EBPdelta (NF-IL6beta). Increased CRH promoter activity following phorbol ester treatment was inhibited by a dominant negative NF-IL6 mutant, showing that the effects of phorbol ester were principally mediated by C/EBP. Moreover, the inverse changes in the expression of CRH in the hypothalamus and spleens of arthritic rats were paralleled by similar inverse changes in NF-IL6beta expression in these organs. These data show that some transcription factors associated with cytokine signalling can also activate the CRH promoter.

Animals↗
PubMed1997

The different inhibitory domains of the Oct-2 transcription factor have distinct functional activities.

The Oct-2 POU family transcription factor contains three distinct regions whose deletion reduces its ability to inhibit transcription via its octamer binding site. Here we show that only one of these inhibitory domains is capable of also inhibiting the activity of activating molecules bound at adjacent sites upstream of a TATA box-containing promoter whereas the other two regions are inactive in this assay. None of the three regions is able to achieve this effect when located upstream of the same promoter containing an initiator motif. The mechanisms of action of these domains and their role in the functioning of the Oct-2 factor are discussed.

Animals↗
PubMed1997

Coordinate induction of the three neurofilament genes by the Brn-3a transcription factor.

The POU domain transcription factor Brn-3a is able to stimulate neurite outgrowth when overexpressed in the neuronal ND7 cell line, whereas the closely related Brn-3b factor does not have this effect. We show that Brn-3a overexpression also enhances the expression of the three neurofilament genes at both the mRNA and protein levels, whereas Brn-3b overexpression has no effect. In addition Brn-3a activates the three neurofilament gene promoters in co-transfection assays in both neuronal and non-neuronal cells. As observed for enhanced neurite outgrowth, the stimulation of neurofilament gene expression and activation of the neurofilament gene promoters is observed with the isolated POU domain of Brn-3a. A single amino acid change in the POU homeodomain of Brn-3a to the equivalent amino acid in Brn-3b abolishes its ability to activate the neurofilament promoters, whereas the reciprocal change converts Brn-3b to an activator of these promoters.

Animals↗
PubMed1997

A single residue within the homeodomain of the Brn-3 POU family transcription factors determines whether they activate or repress the SNAP-25 promoter.

The closely related POU family transcription factors Brn-3a and Brn-3b differ in their effect on a number of different neuronally expressed promoters such as that of the gene encoding the synaptic vesicle component SNAP-25. Thus Brn-3a activates these promoters whilst Brn-3b represses both their basal activity and their activation by Brn-3a. We show here that alterations of a single amino acid at position 22 in the POU-homeodomain from the isoleucine found in Brn-3b to the valine found at the equivalent position in Brn-3a converts Brn-3b from a repressor to an activator of the SNAP-25 gene promoter. The converse mutation in Brn-3a abolishes its ability to activate the SNAP-25 gene promoter and allows it to repress the basal activity of the promoter and its activation by wild type Brn-3a. This is the first time that a single amino acid change has been shown to convert an activator of a naturally occurring promoter to a repressor and vice versa. These results are discussed in terms of the critical role of position 22 in the POU homeodomain in the protein-protein interactions of POU proteins.

Biotransformation↗
PubMed1997

The octamer-binding proteins Oct-1 and Oct-2 repress the HIV long terminal repeat promoter and its transactivation by Tat.

Although the HIV-1 long terminal repeat (LTR) contains four potential binding sites for the octamer-binding protein, Oct-1, which is known to interact with the HIV-1 Tat protein, the effect of the Oct-1 factor on HIV LTR-driven gene expression has not previously been reported. We show here that both Oct-1, and to a lesser extent the related Oct-2 protein, can repress both the basal activity of the HIV-1 LTR and its transactivation by Tat. These effects are still observed with an HIV LTR construct containing only a single octamer-binding site located between the TATA box and the transcriptional start site. The stronger inhibitory effect of Oct-1 on both these promoters is dependent upon its C-terminal region which cannot be effectively replaced by the equivalent region of Oct-2. These effects are discussed in terms of the regulation of HIV LTR activity in different cell types and in response to T-cell activation.

DNA-Binding Proteins↗
PubMed1997

Inhibition of neuronal process outgrowth and neuronal specific gene activation by the Brn-3b transcription factor.

The differentiation of the ND7 neuronal cell line to a nondividing phenotype bearing numerous neurite processes is accompanied by a dramatic increase in the levels of the activating POU family transcription factor Brn-3a and a corresponding fall in the levels of the closely related inhibitory factor Brn-3b. We have previously shown that the artificial overexpression of Brn-3a in these cells can induce neurite outgrowth and the activation of genes encoding synaptic vesicle proteins in the absence of a differentiation-inducing stimulus. Here we show that overexpression of Brn-3b can reduce process outgrowth and synaptic vesicle gene expression following exposure to a stimulus which would normally induce differentiation. These inhibitory effects are abolished by altering a single amino acid in the POU homeodomain of Brn-3b to its equivalent in Brn-3a. The converse mutation in Brn-3a allows it to inhibit process outgrowth in response to a differentiation-inducing stimulus. Hence a single amino acid difference results in these closely related factors having opposite effects and allows the balance between them to regulate differentiation.

Animals↗
PubMed1997

Interleukin 6 activates heat-shock protein 90 beta gene expression.

The levels of the cytokine interleukin-6 (IL-6) and the heat-shock protein hsp90 have both been reported to be elevated in patients with active systemic lupus erythematosus (SLE). We show that hsp90 protein accumulates to increased levels in both HuH7 hepatoma cells and peripheral blood mononuclear cells (PBMCs) treated with IL-6. In PBMCs this effect occurs without induction of the other hsps, paralleling the specific elevation of hsp90 in SLE. IL-6 is able to activate the hsp90 gene promoter directly; this activation can also be achieved by overexpressing either of the transcription factors NF-IL-6 or NF-IL-6 beta whose synthesis is induced by IL-6 treatment. Hence the induction of hsp90 protein accumulation by IL-6 is likely to be dependent on the enhanced activity of the hsp90 beta gene promoter produced by increased levels of NF-IL-6 and/or NF-IL-6 beta. These effects are discussed in terms of the role of hsp90 in the normal immune system and the mechanism of its activation in patients with SLE.

Blotting, Western↗
PubMed1997

The production, binding characteristics and sequence analysis of four human IgG monoclonal antiphospholipid antibodies.

Antiphospholid antibodies (APL) have a notable association with recurrent miscarriages, arterial and venous thrombosis and thrombocytopenia. Analysis of the potential pathogenic effects of such human antibodies has been hampered by the considerable difficulty in producing IgG as opposed to IgM monoclonal immunoglobulins. We have developed four human monoclonal IgG APL (LJ1, AH2, DA3 and UK4) by fusing the peripheral blood lymphocytes of three patients with SLE with a mouse human heteromyeloma cell line, CB-F7. These antibodies bind to a variety of anionic phospholipids, two (LJ1 and AH2) bind total histones but none binds to ssDNA or dsDNA. Binding to beta 2 GPI is non-specific. UK4 alone demonstrates lupus anticoagulant activity. All four have lambda light chains, two are IgG1 (AH2 and UK4) and two are IgG3 (LJ1 and DA3). These APL utilize VH genes present in the fetally restricted repertoire and multiple somatic mutations in the CDR suggest an antigen-driven process. In contrast, there is no restriction in V lambda gene usage and only one lambda chain is extensively mutated. Two clonally related hybridomas were isolated from a single patients. This supports the theory that clonal expansion is the mechanism whereby antigen selects high affinity mutations.

Amino Acid Sequence↗
PubMed1997

Gene delivery to rat enteric neurons using herpes simplex virus-based vectors.

Neurons of the enteric (gut) nervous system can be cultured in vitro and readily survive transplantation into the brain making close connections with host neurons. As such, they could potentially be used to deliver therapeutic gene products to the brain after transduction with appropriate genes in culture. Here the authors report the first example of gene delivery to such cultured neurons using herpes simplex virus based vectors. They show that viruses lacking the immediate early gene encoding ICP27 (which are unable to replicate lytically) can efficiently deliver a marker gene to enteric neurons without producing extensive cellular damage. In contrast, viruses lacking only the viral neurovirulence factor encoded by ICP34.5 are inefficient in gene delivery, and produce extensive cellular damage, although they cannot replicate lytically in enteric neurons. A virus lacking both ICP27 and ICP34.5, however, produces less cellular damage than one lacking only ICP27, and is as efficient in gene transfer, whereas inactivation of VMW65 reduces toxicity further. The identification of this virus as a safe and efficient gene delivery vector for enteric neurons paves the way for the eventual delivery of therapeutic genes and subsequent transplantation of engineered neurons into the CNS.

Animals↗
PubMed1997

Differential regulation of genes encoding synaptic proteins by the Oct-2 transcription factor.

In order to investigate the effect of the Oct-2 POU family transcription factor on the regulation of genes encoding synaptic proteins, we have used cell lines in which the level of Oct-2 has been greatly reduced using an antisense approach. The reduced Oct-2 level results in enhanced expression of SNAP-25 and synapsin I, indicating that the genes encoding these proteins are normally repressed by Oct-2 in neuronal cells. In contrast, no alteration was observed in the levels of the synaptic proteins, synaptophysin and synaptotagmin. Although the neuronal forms of Oct-2 can repress the synapsin I promoter in co-transfection experiments, indicating that they have a direct effect on the expression of this gene, they have no effect on the activity of the SNAP-25 promoter, indicating that the effect of Oct-2 on this gene is likely to be indirect. These effects are discussed in terms of the differential effect of Oct-2 and the related POU family transcription factor Brn-3a, on the promoters of genes encoding different synaptic proteins.

Animals↗
PubMed1997

How can we use our growing understanding of gene transcription to discover effective new medicines?

Although drugs that target gene transcription are in wide therapeutic use, they were all identified on the basis of their effect on a specific biological process, such as inflammation or hormone responses, and were only subsequently shown to target transcription. The recent progress in understanding the mechanism of action of these drugs, and the mechanisms of transcriptional regulation in general, offers hope for a new generation of drugs isolated on the basis of their ability to modulate either the synthesis of transcription factors, the regulation of their activity by ligands or phosphorylation events, their protein-protein interactions or their binding to DNA.

DNA↗
PubMed1997

Transcription factors: an overview.

This special issue of the International Journal of Biochemistry and Cell Biology contains a series of review articles and original papers dealing with the topic of transcription factors. The purpose of this introductory article is to provide an overview of these factors, their mechanism of action, their regulation and the manner in which alterations in them can result in disease.

Animals↗