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N J Gay

Publications and source records attributed to N J Gay.

At least 91 records · Page 5Linked to original sources
PubMed1995

The dorsal protein enhances the biosynthesis and stability of the Drosophila I kappa B homologue cactus.

The cactus and dorsal proteins are Drosophila homologues of mammalian I kappa B cytoplasmic anchor proteins and rel/NF kappa B transcription factors respectively. They are required for the generation of embryonic dorsoventral polarity and probably at later developmental stages for an innate immune response. In this paper we report on the properties of SLDL, a derivative of the SL2 cell line in which dorsal is expressed constitutively. In SLDL cells biosynthesis of cactus protein is stimulated by approximately 4-fold when compared with SL2 cells. Enhanced biosynthesis of cactus protein cannot be explained solely on the basis of increased expression of the cactus gene as the level of the corresponding mRNA is only 2-fold higher than in SL2 cells. On the basis of these findings we propose that free cytoplasmic dorsal protein is able, directly or indirectly to stimulate translation of the cactus mRNA. Such an arrangement would enable the dorsal protein to be buffered in the cytoplasm of the resting cell over a wide range of concentrations. We also show here that subsequent to biosynthesis the cactus protein is either rapidly degraded or incorporated into complexes with dorsal. Protein that does not associate with dorsal has a half-life of approximately 40 min whereas that which is incorporated into complexes is very stable, having a half life in excess of 24 h. The complexed cactus protein is acted on by protein kinases which generate distinct phophorylated isoforms.

Animals↗
PubMed1995

Wild type and constitutively activated forms of the Drosophila Toll receptor have different patterns of N-linked glycosylation.

Toll is a Drosophila membrane protein related in sequence to the mammalian platelet glycoprotein 1B and to the interleukin-1 receptor. It mediates a signal transduction pathway leading to the development of dorsoventral polarity in the Drosophila embryo. In this paper we show that a constitutively activated mutant receptor, Toll10B, is processed into a distinct isoform of slower electrophoretic mobility when compared with the wild type molecule in both cell lines and the embryo. The wild type protein can also be processed into this form if over-expressed but in the embryo is present as the smaller species. We show that the decrease in the mobility of Toll10B and over-expressed wild type receptors is caused by altered patterns of N-linked glycosylation and that both forms are secreted to the cell surface. On the basis of these results, we propose that the Toll10B receptor is unable to associate with a limiting co-factor which when bound directly or indirectly masks supplementary N-linked glycosylation sites.

Animals↗
PubMed1995

Predicting the impact of measles vaccination in England and Wales: model validation and analysis of policy options.

Measles incidence in England and Wales has fallen to an all-time low. Attention is now focused on preventing local outbreaks, and, in the long run, on the elimination of indigenous measles. A realistic age-structured (RAS) mathematical model of measles transmission is used to reconstruct the impact of measles vaccination in England and Wales from 1968 to the present and to evaluate the merits of future policy options. In general, the predictions of the model show good agreement with long-term age stratified case reports and seroprevalence surveys. The model underestimates the proportion of cases that are notified in 0-2-year-old children. However, recent work suggests a high degree of misdiagnosis in this age group. Projections on the basis of the existing vaccination strategy in the UK suggest that the present level of measles vaccine coverage will be insufficient to eliminate small seasonal outbreaks of measles. This result is, however, sensitive to the assumed level of vaccine efficacy. Explorations of a variety of changes to current vaccination strategy favour a 2-dose schedule with the second dose administered at age 4 years irrespective of vaccination history. A vaccination campaign in school-age children, to reduce deficits in herd immunity, would accelerate progress towards measles elimination.

Adolescent↗
PubMed1995

Interpretation of serological surveillance data for measles using mathematical models: implications for vaccine strategy.

Serological surveillance of measles immunity has been carried out in England since 1986/7. Results from sera collected in 1989-91 revealed that the proportion of school age children who were susceptible to measles was increasing, following the introduction of the measles, mumps and rubella vaccination programme in October 1988. Mathematical models are used to interpret these data and determine whether this increasing susceptibility is sufficient to allow a resurgence of disease from the low levels achieved by 1993. The models summarize serological profiles by a single parameter, the reproduction number R, which quantifies the level of herd immunity in the population. Results showed that there was cause for concern over the levels of susceptibility to measles, with an epidemic of over 100,000 cases likely in 1995/6. These predictions are consistent with trends in the incidence and age distribution of measles and have enabled the planning of a major vaccination campaign.

Adolescent↗
PubMed1995

Hand-genital transmission of genital warts? An analysis of prevalence data.

The role of hand-genital transmission in the aetiology of genital warts is unclear. However this route is suggested by a number of observations including the relatively high proportion of genital warts in children which contain HPV types 1-4 (15% for children and 2% for adults). We compared two transmission models; one which assumes that hand-genital transmission occurs and one that it does not, and determined the conditions in which each model can reflect the available prevalence data. Hand-genital transmission provides a simple explanation of the observed differences in the proportions of genital warts containing HPV types 1-4 and 6/11 in children and adults. If hand-genital transmission does not occur, the observed difference could only be explained by an eightfold greater probability of transmission to children of types 1-4 than types 6/11, or by an eightfold greater duration of infection with types 1-4. Our findings provide support for the view that genital warts may be transmitted by hand-genital contact.

Adult↗
PubMed1994

Age specific antibody prevalence to parvovirus B19: how many women are infected in pregnancy?

Infection with parvovirus B19 is an important cause of late fetal mortality in the second trimester, and many infections in pregnancy remain undiagnosed. A serological survey stratified by age has been used to estimate the incidence of maternal infection with parvovirus B19 in pregnancy. Serum remaining from specimens submitted for diagnosis from 6864 people of all ages to seven public health laboratories in England was tested for antibody to parvovirus B19. The antibody prevalence rose with age to 45% at 10 years and 60% to 70% in adults. The age specific force of infection was highest in children aged less than 10 years and lowest in adults. Maternal infection with parvovirus B19 is estimated to occur in approximately one pregnancy in 400. It has been estimated that fetal death occurs in 9% of these cases, which suggests that parvovirus B19 may cause more than 150 fetal deaths in England and Wales each year. Testing for evidence of recent infection with parvovirus B19 should be considered for unexplained cases of fetal hydrops in the second trimester, especially in years of parvovirus B19 epidemics.

Adolescent↗
PubMed1994

Selective rubella vaccination programmes: a survey of districts in England and Wales.

A survey of district immunisation coordinators in the 183 health districts of England and Wales was carried out to assess the implementation of selective rubella vaccination programmes. The survey showed that school health services vaccinate schoolgirls against rubella in 161 (93%) of the 173 districts whose immunisation coordinators responded. The accuracy of data on vaccination coverage of schoolgirls is limited because districts interpret the numerator and denominator required by the Department of Health in different ways. Districts have also experienced problems in monitoring the uptake of vaccination through general practitioners. Comprehensive antenatal screening programmes for rubella immunity operate in 164 districts (95%), but few districts are auditing the postpartum vaccination of seronegative women. If child health computer records were maintained and updated until children left school, it would be possible to produce reliable data on the percentage of girls who had been vaccinated against rubella by the age of 14.

Adolescent↗
PubMed1994

Sequence and expression of LRR47, a novel embryonic leucine rich repeat protein of Drosophila.

Leucine-rich repeats (LRRs) are 22-28 amino acid long sequence motifs found in a variety of extracellular, membrane and cytoplasmic proteins. They are believed to mediate specific protein-protein interactions and to function in cellular adhesion. In Drosophila, four LRR proteins are known and each plays an important role in embryogenesis. In this paper we report the cloning of a cDNA that encodes a fifth Drosophila embryonic LRR protein, LRR47. The sequence includes a hydrophobic N-terminal which may constitute an ER signal sequence, eight LRR copies and a unique C-terminal. The transcript of the LRR47 gene is detected in adult females and in early embryogenesis. It is not found in adult males and is only present at low levels in embryos after 6 h of development. In Western blot experiments, a protein of approx. 47 kDa, which is expressed in a similar developmental profile and purifies in peripheral membrane protein extracts, is detected by an antibody specific for LRR47. The LRR47 gene maps to position 32A on the left arm of chromosome 2, an interval in which three genes with semi-lethal maternal effects (dal, hup and wdl) are located.

Amino Acid Sequence↗
PubMed1994

Drosophila ribosomal protein L18a: cDNA sequence, expression and chromosomal localization of the gene.

We describe the cDNA sequence of the Drosophila homologue of the rat ribosomal protein L18a. The protein sequence predicted has identical or conservatively substituted amino acids in 80% of positions. It is distinctly basic in character with an overall net positive charge of + 20. Analysis of L18a RNA with the Northern blot technique shows it to be expressed both during embryonic development and in the adult fly. In situ hybridisation to polytene chromosomes reveals that the L18a gene(s) is located at 54B on the second chromosome.

Amino Acid Sequence↗
PubMed1994

Age-specific antibody prevalence to hepatitis A in England: implications for disease control.

Sera from an age-stratified sample of 7196 individuals, submitted for diagnostic purposes to four public health laboratories in England in 1986/7, were tested for hepatitis A antibody. The serological profiles, which showed marked regional differences, were consistent with declining incidence in the past. The decline in the incidence of hepatitis A has resulted in an increase in susceptibility in adults. This has three main consequences: an increase in the average age of infection may be leading to an increase in morbidity; normal immunoglobulin may become less protective against hepatitis A; the risk of transmission through blood products contaminated by viraemic blood donors may rise. Current average annual incidence in 5-14-year olds was estimated to vary between regions from 0.5-1.9%. This supports the view that, in the absence of a vaccination programme, hepatitis A will remain endemic unless there are further improvements in living conditions and standards of hygiene. A vaccine giving long-lasting protection could eliminate hepatitis A transmission with modest coverage at a young age. Targeting childhood vaccination on economically deprived areas or using vaccine to control outbreaks might be more effective policies.

Adolescent↗
PubMed1993

The Drosophila ankyrin repeat protein cactus has a predominantly alpha-helical secondary structure.

The cactus protein is the Drosophila homologue of the mammalian I kappa B family of cytoplasmic anchor proteins. We have expressed in E. coli and purified a cactus fusion protein, CACT-Bgl. CACT-Bgl protein contains the six ankyrin repeat sequences which are necessary for specific binding to the Drosophila rel family transcription factor dorsal. We show that the purified CACT-Bgl protein can bind specifically to dorsal and, using circular dichroism spectroscopy, that the protein adopts a largely alpha-helical secondary structure. A further analysis of the ankyrin repeat domains of cactus, using an improved secondary structure prediction program indicates that the N-terminal of the repeat will form into a loop structure and the C-terminal section into an interrupted, amphipathic alpha-helix. On the basis of these findings we propose that the ankyrin repeats of cactus fold together into helical bundles interconnected by diverged loops.

Amino Acid Sequence↗
PubMed1993

Relocalization of Drosophila dorsal protein can be induced by a rise in cytoplasmic calcium concentration and the expression of constitutively active but not wild-type Toll receptors.

The generation of dorso-ventral polarity in Drosophila relies on the formation of a nuclear gradient of the rel/nuclear factor kappa B transcription factor dorsal in the pre-cellular syncitial embryo by a process of differential nuclear localization. It is thought that the gradient is formed by activation at ventral positions of the membrane receptor Toll that in turn causes the local dissociation of dorsal from the cytoplasmic anchor protein cactus. Although Toll is related in its cytoplasmic domain to the interleukin-1 receptor little is known about the signal transduction pathways that lead from Toll to the relocalization of dorsal. In this paper we have used immunofluorescence microscopy as a direct assay of dorsal protein nuclear localization in the Drosophila cell line Schneider 2. We find that increased cytoplasmic calcium concentration and the expression of constitutively active Toll receptors can induce the relocalization of dorsal. By contrast, we find that activation of endogenous protein kinase A and expression of wild-type Toll receptors, which activate zen-chloramphenicol acetyltransferase reporter genes in this system, have only a marginal effect on the cellular distribution of the dorsal protein. Treatment of cells with activators of protein kinase C and radical oxygen intermediates, both of which activate nuclear factor kappa B, also has little effect on dorsal protein localization. We propose that different threshold levels of dorsal activation can be established by distinctly regulated signal transduction pathways.

Androgen-Binding Protein↗
PubMed1992

Regulation of translation and proteolysis during the development of embryonic dorso-ventral polarity in Drosophila. Homology of easter proteinase with Limulus proclotting enzyme and translational activation of Toll receptor synthesis.

The generation of dorso-ventral polarity during Drosophila embryogenesis is regulated by the action of 12 maternally expressed gene products, the dorsal group. These products act together to form a dorso-ventral nuclear gradient of the transcription factor dorsal. At least three of the dorsal group genes (snake, easter and gastrulation defective) encode secreted serine proteinases which probably function during early development in the perivitelline compartment of the embryo. Here, we report that the easter proteinase is homologous in its light chain sequence to the haemocyte proclotting enzyme (PCE) of the Japanese horseshoe crab Tachypleus tridentatus. PCE is the terminal member of a proteolytic cascade activated in response to microbial polysaccharides and acts to cleave coagulogen, an invertebrate equivalent of fibrinogen. On the basis of this homology we are able to predict with confidence the overall primary structure of the easter proteinase, its mode of activation and its substrate specificity. The result also suggests that easter functions zygotically in haemocytes in a Drosophila defence response analogous to that found in Tachypleus. We also show here that the Toll receptor protein is absent in early cleavage embryos but accumulates rapidly at the syncitial blastoderm stage, the developmental stage at which its function is required. This finding suggests that translation of Toll mRNA is regulated in response to fertilisation and egg deposition. These two observations are consistent with a model of dorso-ventral pattern formation in which a proteolytic cascade is activated uniformly in the perivitelline compartment of the embryo and causes the release of ventrally localised ligands of the Toll receptor. A possible alternative model in which a proteolytic cascade is activated in response to a ventrally restricted signal is also discussed.

Amino Acid Sequence↗
PubMed1991

A leucine-rich repeat peptide derived from the Drosophila Toll receptor forms extended filaments with a beta-sheet structure.

Leucine-rich repeats (LRRs) are 22-28 amino acid-long sequence motifs found in a family of cytoplasmic, membrane and extracellular proteins. There is evidence that LRRs function in signal transduction, cellular adhesion and protein-protein interactions. Here we report unusual properties of a synthetic LRR peptide derived from the sequence of the Drosophila membrane receptor Toll. In neutral solution the peptide forms a gel revealed by electron microscopy to consist of extended filaments approximately 8 nm in thickness. As the gel forms, the circular dichroism spectrum of the peptide solution changes from one characteristic of random coil to one associated with beta-sheet structures. Molecular modelling suggests that the peptide form an amphipathic structure with a predominantly apolar and charged surface. Based on these results, models for the gross structure of the peptides filaments and a possible molecular mechanism for cellular adhesion are proposed.

Amino Acid Sequence↗
PubMed1990

Spatial and temporal distribution of a Drosophila melanogaster embryonic variable nuclear antigen.

A polyclonal antibody has been prepared that specifically recognises a nuclear protein antigen in Drosophila embryos. During development, the antigen appears initially to be uniformly distributed but by nuclear division cycle 10 is seen to accumulate in nuclei in a manner suggesting that it is destroyed or becomes modified upon transition from S- to M phase of the nuclear division cycle. This conclusion is supported by the observed disappearance of the antigen from the postblastoderm nuclei in a manner that reflects the pattern of the first asynchronous postblastoderm cell division and persistence in the polyploid nuclei of the amnioserosa that do not undergo further cell or nuclear division. In Western blot experiments, the antibody detects specifically a 105 kDa nuclear protein that probably corresponds to the antigen detected in embryos by immunocytochemical means.

Animals↗
PubMed1990

The Drosophila membrane receptor Toll can function to promote cellular adhesion.

The product of the Toll gene is a membrane protein required for the formation of dorso-ventral polarity during early embryogenesis in Drosophila melanogaster. It acts together with the other dorsal group gene products to specify a nuclear gradient of dorsal morphogen in the syncytial blastoderm stage embryo. Here we report the presence in Toll protein of additional sequences held in common with the human membrane receptor platelet glycoprotein 1b (Gp1b). We propose that these sequences in Toll form disulphide linked extracellular domains that are important for the binding of ligands in the perivitelline space of the embryo. In addition, we show that expression of Toll protein induced in a non-adhesive cell line promotes cellular adhesion, a property held in common with the related Drosophila glycoprotein chaoptin. Toll protein in such aggregates accumulates at sites of cell-cell interaction, a characteristic displayed by other cellular adhesion molecules. Taken together these findings suggest that the biochemical function of Toll protein is more closely analogous to that of Gp1b than previously thought.

Amino Acid Sequence↗