Biomedical subjects

D S Latchman

Publications and source records attributed to D S Latchman.

At least 163 records · Page 9Linked to original sources

PubMed1997

Apoptosis in peripheral lymphocytes in systemic lupus erythematosus: a review.

There is increasing evidence to suggest that abnormalities in apoptosis may play a part in the pathogenesis of systemic lupus erythematosus (SLE). For example, there is now considerable evidence that bel-2 expression is enhanced in a proportion of peripheral T cells, but not in B cells, in SLE patients and correlates with overall disease activity regardless of the activity index employed. Further work is required to establish whether enhanced bel-2 expression by some T cells in SLE patients is related to their activation or intrinsically enhanced by genetic predisposition. Mutations in Fas result in a lymphoproliferative syndrome and may play a role in accelerating autoimmune disease. A report of three children with mutations in Fas has once again focused attention on this regulator of apoptosis. The relationships between inducers and inhibitors of apoptosis may differ in different cell types, and must be elucidated before the implications of observations made in lymphocytes from SLE patients can be fully understood.

Animals↗

PubMed1997

The Brn-3a transcription factor induces neuronal process outgrowth and the coordinate expression of genes encoding synaptic proteins.

The Brn-3a POU family transcription factor is expressed only in posmitotic neurons in the central nervous system and identifies the first differentiated neurons to appear in the midbrain, hindbrain, and spinal cord during development. This factor is also induced when undifferentiated proliferating ND7 cells cease dividing and differentiate to a mature neuronal-like phenotype bearing numerous neurite processes. We show that overexpression of Brn-3a in undifferentiated ND7 cells induces a mature neuronal phenotype characterized by process outgrowth and the induction of genes encoding synaptic proteins, although the cells continue to proliferate. In contrast, the closely related factors Brn-3b and Brn-3c do not have this effect. Although the N-terminal activation domain of Brn-3a is required for maximum induction of neurite outgrowth and gene expression, these effects are primarily dependent on the DNA binding POU domain, which also acts as an activation domain. Overexpression of the isolated POU domain of Brn-3a is sufficient to induce neurite outgrowth, while the ability of full-length Brn-3a to do so is abolished by mutating a single amino acid in the Brn-3a POU homeodomain to its equivalent in Brn-3b. Thus, Brn-3a appears to play a critical role in the specification of the mature neuronal phenotype, acting by stimulating the expression of genes whose products are required for process outgrowth and synapse formation.

Animals↗

PubMed1997

Elevated soluble fas production in SLE correlates with HLA status not with disease activity.

Evidence from animal models of lupus suggests that disruption of Fas-mediated apoptotic events may play a role in systemic lupus erythematosus (SLE). The recently described secreted from of Fas (sFas) could interfere with apoptotic events by blockading Fas/Fas ligand interactions. We describe elevated secreted Fas protein in sera from 60 patients with SLE compared with controls but neither sFas protein nor sFas mRNA levels correlated with disease activity. At the mRNA level there is strong evidence that individuals with human leucocyte antigens common in SLE patients have a genetic predisposition for increased secreted Fas production.

Adult↗

PubMed1996

Differential regulation of genes encoding synaptic proteins by members of the Brn-3 subfamily of POU transcription factors.

The three members of the Brn-3 subfamily of POU transcription factors have distinct effects on target gene expression. We show that the promoter of the gene encoding the presynaptic nerve terminal protein SNAP-25 resembles previously characterised target genes in being activated by Brn-3a and Brn-3c, but being repressed by Brn-3b. Unlike other target genes, however, the SNAP-25 promoter can be activated by either the N- or C-terminal activation domains of Brn-3a. In contrast to the SNAP-25 gene, the gene encoding the synaptic vesicle protein synapsin 1 is activated by all the Brn-3 factors, the first gene for which this activation pattern has been reported Interestingly, however, similar activation by all three Brn-3 factors can be observed if the SNAP-25 promoter is truncated by removal of sequences from -2200 to -288 relative to the transcriptional start site. Moreover, a region of the SNAP-25 promoter from -283 to -126 can render a heterologous promoter responsive to activation by all three Brn-3 factors. Differences in promoter structure may thus result in differences in the response to different Brn-3 factors, thus allowing these factors to produce diverse activation patterns of neuronally expressed genes, such as those encoding different synaptic proteins.

Animals↗

PubMed1996

Differential regulation of the two neuronal nitric-oxide synthase gene promoters by the Oct-2 transcription factor.

The Oct-2 transcription factor has been shown previously to repress both the cellular tyrosine hydroxylase and the herpes simplex virus immediate-early genes in neuronal cells. Here we identify the gene encoding the neuronal nitric-oxide synthase (nNOS) as the first example of a gene activated in neuronal cells by Oct-2. The levels of the nNOS mRNA and protein are greatly reduced in neuronal cell lines in which Oct-2 levels have been reduced by an antisense method, although these cells have enhanced levels of tyrosine hydroxylase. Moreover, the nNOS gene regulatory region is activated by Oct-2 expression vectors upon cotransfection into both neuronal and non-neuronal cells, and this response is dependent upon a 20-amino acid region within the COOH-terminal activation domain of Oct-2. Of the two closely linked promoters that drive nNOS gene expression, only the downstream 5.1 promoter is activated by Oct-2, whereas the 5.2 promoter is unaffected. These effects are discussed in terms of the potential role of Oct-2 in regulating nNOS expression in the nervous system.

Animals↗

PubMed1996

Increased heat shock protein 90 (hsp90) expression leads to increased apoptosis in the monoblastoid cell line U937 following induction with TNF-alpha and cycloheximide: a possible role in immunopathology.

In this study, we examined the hypothesis that heat shock proteins (hsp) (such as hsp72 and hsp90) are implicated in the regulation of forms of cell injury that lead to programmed cell death. The monoblastoid cell line U937 has been used as a model system. For hsp90, which is not heat inducible in this cell line, we used stable U937 transfectants that either hyperexpress or hypoexpress the protein. For hsp72 (which is reproducibly induced in all three cell lines to relatively high levels of expression), we studied U937 cells before and after heat shock. We showed that apoptosis does occur in the monoblast/mononuclear phagocyte lineage, and that it could be induced in vitro by serum deprivation, UV light, or TNF-alpha in combination with cycloheximide (cx). However, an excess of hsp90 is associated with increased apoptosis when the cells are treated with a combination of TNF-alpha and cx but not when they are exposed to UV B radiation. This was complemented by the finding that reduced hsp90 levels correlate with protection against apoptosis in the TNF-alpha- and cx-treated cells. Furthermore, new synthesis of hsp72 does not protect against apoptosis. Thus, hsp90 levels may play a role in controlling the part played by mononuclear phagocytes in immunopathology.

Antigens, CD↗

PubMed1996

The functionally antagonistic POU family transcription factors Brn-3a and Brn-3b show opposite changes in expression during the growth arrest and differentiation of human neuroblastoma cells.

The Brn-3a and Brn-3b POU family transcription factors have previously been shown to have functionally antagonistic effects, with Brn-3a activating specific gene promoters which are repressed by Brn-3b. We show here that proliferating cell lines of human neuroblastoma origin have a high ratio of Brn-3b to Brn-3a compared with other cell lines derived from related tumours. Moreover, the level of Brn-3a rises and that of Brn-3b falls when neuroblastoma cell lines are exposed to treatments which induce a cessation of proliferation. Such treatments also result in the activation of a test promoter which is normally stimulated by Brn-3a and repressed by Brn-3b. Our findings suggest that these antagonistic factors may play a key role in regulation of gene expression during human neuroblastoma differentiation and could thus represent a potential therapeutic target for treatments designed to induce such differentiation.

Base Sequence↗

PubMed1996

The ability of the inhibitory domain of the POU family transcription factor Oct-2 to interfere with promoter activation by different classes of activation domains is dependent upon the nature of the basal promoter elements.

The Oct-2 transcription factor contains an inhibitory domain which is able to repress transcription following DNA binding. Here we show that within the neuronally expressed Oct-2.5 form, the inhibitory domain can strongly inhibit activation by transcription factor activation domains which are either composed predominantly of acidic residues or contain the HOB motif, whereas it has a weaker effect or no effect on proline-rich activation domains and on a glutamine-rich domain. In contrast, the isolated inhibitory domain of Oct-2 can efficiently repress all types of activation domains. This effect is observed however, only on TATA box-containing promoters and not on promoters containing an initiator motif. This widespread inhibition of different activation domains and its dependence on the nature of the basal promoter elements indicate that the inhibitory domain is likely to act by contacting a common downstream target of activation domains within the basal transcriptional complex bound at the TATA box rather than quenching specific activation domains by direct interaction. These effects are discussed in terms of the functional role of the inhibitory domain within Oct-2.5 and the mechanism by which it acts.

Animals↗

PubMed1996

The role of the 90-kDa heat shock protein in cell cycle control and differentiation of the monoblastoid cell line U937.

The human monoblastoid cell line, U937, has been widely used to study proliferation and differentiation in the monocyte-macrophage lineage. Recent evidence from other cell systems suggests that heat shock proteins (hsps) may participate in these processes. Therefore, we have examined expression of hsp and the effect of either increased or decreased expression of the hsp90 in U937 cells. Parental U937 cells express high levels of hsp90, hsp73, and hsp65, but little hsp72. Heat shock at 42 degrees C for 30 min increased hsp72 levels but caused no change in hsp90. U937 cells transfected with the expression vector pBA.4 containing either an anti-sense or a sense hsp90 cDNA insert showed constitutive decrease, or increase, in expression of hsp90. Decreased hsp90 levels slowed the rate of cell division and levels of hsp90 correlated both with the responses to phorbol esters and with phenotypic changes: antisense-transfected cells expressed less CD50. Sense-transfected cells showed no change in cell cycle, but expressed less CD14 than controls. Thus, hsp90 plays a role in the monocyte-macrophage lineage, participating in proliferation and cell cycle control and in the acquisition of functional heterogeneity of the mature macrophage phenotype, with potential effects on the role of the macrophage in innate immunity.

Antigens, CD↗

PubMed1996

Activation of the gene encoding decay accelerating factor following nerve growth factor treatment of sensory neurons is mediated by promoter sequences within 206 bases of the transcriptional start site.

Using two independent differential screening procedures designed to identify novel mRNAs induced by nerve growth factor (NGF) treatment of adult dorsal root ganglion (DRG) neurons, we have isolated cDNA clones derived from the gene encoding decay accelerating factor (DAF). Hybridization analysis and semi-quantitative polymerase chain reaction confirmed that the DAF mRNA was indeed induced in NGF-treated adult DRG neurons. Moreover, the DAF gene promoter is NGF inducible (approximately two- to threefold) when transfected into DRG neurons, and this effect is primarily dependent on sequences between -206 and -77 relative to the transcriptional start site. Hence, the DAF gene constitutes a novel NGF-inducible gene whose mRNA is elevated in response to NGF treatment of DRG neurons. The potential significance of this effect is discussed in terms of the role of NGF in modulating the transcriptional activity and function of adult DRG neurons.

Animals↗

PubMed1996

Differential regulation of neuronal nicotinic acetylcholine receptor subunit gene promoters by Brn-3 POU family transcription factors.

The regulatory region of the neuronal nicotinic acetylcholine (nACh) receptor alpha 2 subunit gene is activated by the Brn-3b POU family transcription factor but not by the closely related factors Brn-3a and Brn-3c. This pattern of regulation has not previously been observed for other neuronally expressed genes, several of which, such as those encoding alpha-internexin or SNAP-25, are activated by Brn-3a and Brn-3c but repressed by Brn-3b. The alpha 3 nACh receptor subunit gene is also shown to be activated by Brn-3a but is repressed by Brn-3b and Brn-3c. In contrast, the Brn-3 POU family transcription factors have no effects on either the alpha 7 or beta 4 nACh receptor subunit genes. The actions of Brn-3b on the alpha 2 subunit are thus in contrast to the inhibitory actions of Brn-3b on several promoters that are activated by Brn-3 alpha. The different actions of the Brn-3 POU factors on the range of nACh receptor genes tested suggests that the novel stimulation of the alpha 2 subunit by Brn-3b is specific to this subunit and not a general feature of nACh receptor genes.

Acetylcholine↗

PubMed1996

A single amino acid change converts an inhibitory transcription factor into an activator.

The closely related POU family transcription factors Brn-3a and Brn-3b differ in their functional activity with Brn-3a activating several target promoters, which are repressed by Brn-3b. Brn-3b also prevents promoter activation by Brn-3a. Here we have altered a single isoleucine residue in the POU homeodomain of Brn-3b to the valine residue found at the equivalent position in Brn-3a. This change not only abolishes the ability of Brn-3b to repress basal and Brn-3a-stimulated promoter activity but also converts it to an activator of similar potency to Brn-3a. Hence a single amino acid difference determines the difference between an activator and a repressor in the Brn-3 family.

Base Sequence↗

PubMed1996

Oct-1 [corrected] and Oct-2 DNA-binding site specificity is regulated in vitro by different kinases.

The transcription factors Oct-1 and Oct-2 bind differentially to three octamer binding sequences corresponding to the octamer binding site from the H2B promoter [ATGCTAATAA], a simple TAATGARAT motif, found in herpes simplex virus IE4/5 genes [GCGGTAATGAGAT], and a perfect consensus overlapping octamer/TAATGARAT motif [ATGCTAATGAGAT]. By comparing the effects of protein kinase A, protein kinase C and casein kinase 2 in vitro on the binding of Oct-1 and Oct-2 to the three motifs, we show that the actions of these kinases regulate Oct-1 and Oct-2 DNA binding independently of each other in a binding-site-specific manner. Inhibition of cellular phosphatases also regulate Oct-1 and Oct-2 DNA binding in a binding-site-specific manner. Both kinase and phosphatase activity are important for regulating the DNA binding activity of Oct-1 and Oct-2 because, in the presence of phosphatase inhibitors, protein kinase A attenuates the binding of both Oct-1 and Oct-2 to the octamer binding site but enhances binding when phosphatase inhibitors are omitted. Thus the DNA specificity of Oct-1 and Oct-2 can be regulated in vitro by the action of different kinases.

Base Sequence↗

PubMed1996

Activation and repression of gene expression by POU family transcription factors.

The POU family transcription factors Oct-2 and Brn-3 utilize different mechanisms to produce a variety of effects on gene expression particularly in the nervous system. In the case of Oct-2, a single gene produces a primary RNA transcript. This transcript then undergoes alternative splicing to yield a variety of different mRNAs encoding Oct-2 isoforms which either activate or repress gene expression. In contrast, three distinct genes encode the closely related Brn-3 factors, Brn-3b, Brn-3b and Brn-3c. Although the proteins encoded by the Brn-3a and Brn-3c genes activate their target genes Brn-3b represses these genes and can also interfere with activation by Brn-3a or c. These finding indicate that diverse mechanisms are used to generate activating or repressing forms of POU family transcription factors.

Amino Acid Sequence↗

PubMed1996

The different activities of the two activation domains of the Brn-3a transcription factor are dependent on the context of the binding site.

The POU (Pit-Oct-Unc) family transcription factor Brn-3a contains two distinct activation domains, one at the N terminus of the molecule and one at the C terminus coincident with the DNA binding domain. These different activation domains have been shown previously to differ in their ability to activate an artificial test promoter containing a Brn-3a binding site and the naturally occurring alpha-internexin gene promoter. Here we identify the target site for Brn-3a in the alpha-internexin gene promoter and show that it can confer responsiveness to Brn-3a on a heterologous promoter. One of the single-stranded DNA sequences derived from either this novel Brn-3a binding site or from the previously characterized site in the test promoter are shown to bind Brn-3a preferentially compared with the complementary single strand or the corresponding double-stranded sequence. The pattern of responsiveness of these two sequences when cloned upstream of the same test promoter and co-transfected with constructs encoding various portions of Brn-3a indicates that the activity of the two Brn-3a activation domains is dependent upon differences in the context of the target sequence in each promoter rather than on differences in the target sequence itself.

Animals↗

PubMed1996

Specific up-regulation of the POU domain transcription factor Oct-2 following axotomy.

Peripheral nerve damage causes a dramatic alteration to the gene expression in primary sensory neurons, changes within the neuronal cell body giving rise to an altered phenotype, adapted for axonal regeneration. Such changes suggest an alteration in activity, or levels, of cellular transcription factors. The POU family transcription factor Oct-2 is known to be induced in sensory neurons by nerve growth factor (NGF) and might therefore be affected by the removal of target-derived NGF following axotomy. Paradoxically, however, the expression of Oct-2 showed a transient increase of two- to three-fold 24 h after axotomy. In contrast, axotomy had no effect on the levels of the Brn-3 sub-family of POU proteins, indicating that this effect was specific for Oct-2.

Animals↗

PubMed1996

Over-expression of heat shock protein 70 protects neuronal cells against both thermal and ischaemic stress but with different efficiencies.

Primary cultures of dorsal root ganglia (DRG) sensory neurons can be protected against subsequent severe thermal or ischaemic stress by prior exposure to a mild thermal or ischaemic insult. The degree of protection correlates with the amount of 70 kDa heat shock protein (hsp70) induced by the mild stress. We show directly that over-expression of hsp70 alone is sufficient to protect DRG neurons against thermal or ischaemic stress with a given level of hsp70 over-expression providing greater protection against thermal stress. In contrast over-expression of the 90 kDa heat shock protein (hsp90) has little or no protective effect against either stress. These results are discussed in terms of the role of individual hsps in protecting neuronal cells against different stresses.

Brain Ischemia↗

PubMed1996

The ability of POU family transcription factors to activate or repress gene expression is dependent on the spacing and context of their specific response elements.

The different forms of the Oct-2 and Brn-3 POU family transcription factors can have distinct effects on their target promoters involving both the activation and repression of gene expression. To investigate the requirements for these effects we have altered both the context and spacing of the two TAATGARAT binding sites for these factors within the herpes simplex virus immediate-early 3 gene promoter. We show that the activation of this promoter by Brn-3a and its repression by Brn-3b is dependent on the correct spacing of these binding sites. In contrast, repression of the promoter by Oct-2.4 and Oct-2.5 is dependent on both the spacing and context of these sites with the requirements for repression by Oct-2.4 or Oct-2.5 being different. These effects are discussed in the context of the mechanisms by which POU factors activate or repress their target genes.

Animals↗