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W Ding

Publications and source records attributed to W Ding.

At least 37 records · Page 2Linked to original sources
PubMed2001

Human T-lymphotropic virus type 1 p30(II) regulates gene transcription by binding CREB binding protein/p300.

The highly conserved coadapters CREB binding protein (CBP) and p300 form complexes with CREB as well as other DNA binding transcription factors to modulate chromatin remodeling and thus transcription. Human T-lymphotropic virus type 1 (HTLV-1) transcription is controlled, in part, by the CREB/ATF family of transcription factors which bind promoter sequences and function as complexes with the viral oncogenic protein Tax. We have reported that the nuclear localizing protein p30(II) of HTLV-1 functions as a transcription factor, differentially modulates CREB-responsive promoters, and is critical for maintenance of proviral loads in rabbits. In this study, we tested whether p30(II) directly interacts with CBP/p300 to modulate gene transcription. Gal4(BD)-p30(II)-mediated transactivation was enhanced following exogenous expression of p300 and was competitively repressed by the p300 binding protein, adenovirus E1A, and E1ACR2 (mutated for retinoblastoma binding but retaining p300 binding). In contrast, E1ACR1 (mutated for p300 binding) failed to alter Gal4(BD)-p30(II)-mediated transactivation. In addition, Gal4(BD)-p30(II)-mediated transactivation was competitively inhibited by the cotransfection of CMV-p30(II)-HA and CMV-Tax but could be rescued by exogenous p300. Importantly, we demonstrate that p30(II) colocalizes with p300 in cell nuclei and directly binds to CBP/p300 in cells. Deletion mutants of CBP/p300 were used to localize the site critical for binding p30(II) to a highly conserved KIX region. DNA binding assays confirmed the interference of p30(II) with the assembly of CREB-Tax-p300/CBP multiprotein complexes on 21-bp repeat oligonucleotides in vitro. Collectively, our results demonstrate that CBP/p300 is a cellular protein target for HTLV-1 p30(II) and mediates its transcriptional effects in vivo.

Adenovirus E1A Proteins↗
PubMed2001

Senescence represses the nuclear localization of the serum response factor and differentiation regulates its nuclear localization with lineage specificity.

The differentiation of cultured 3T3T mesenchymal stem cells into adipocytes represses growth factor responsiveness by limiting the nuclear localization of the serum response factor (SRF) that binds to and activates the promoters of growth control genes that contain the serum response elements (SRE), such as junB and c-fos. The regulation of SRF nuclear localization by adipocyte differentiation is specific, because we show that adipocyte differentiation does not repress the nuclear localization of six other transacting factors. To determine if repression of growth factor responsiveness that occurs during senescence also represses the nuclear localization of SRF, we studied normal human WI-38 fibroblasts at low versus high population doublings. The results show that SRF localizes to the nucleus of proliferative cells whereas in senescent cells SRF can not be detected in the nucleus. This result is apparent in both immunofluorescence assays and in western blot analysis. We next evaluated the cellular distribution of SRF in selected human tissues to determine whether the loss of proliferative potential in vivo could have a different effect on SRF nuclear localization. We found that in cells of the small bowel mucosa, differentiation modulates SRF nuclear localization in an opposite manner. Minimal SRF expression and nuclear localization is evident in undifferentiated cells at the base of crypts whereas increased SRF expression and nuclear localization is evident in differentiated cells at the surface tip of the villus. These results together establish that regulation of SRF expression and nuclear localization is important in senescence and differentiation in a lineage specific manner.

Adipocytes↗
PubMed2001

Effect of long-term treatment with vanadate in drinking water on KK mice with genetic non-insulin-dependent diabetes mellitus.

The glucose-lowering effect of vanadate, ammonium metavanadate (AMV), on diabetic KK mice was examined. Five-week-old male KK mice were administrated with a solution of AMV via drinking water at concentrations of vanadium (V) with 0.1, 1.0, 10 and 100 microg/mL for a period of 10 wk, respectively. Body weight, consumption of food and water, and blood glucose levels was measured every week for 10 wk. The results showed that food consumption and body weight in the experimental groups were similar to those in the control group. A statistically significant decrease of drinking water consumption and blood glucose levels in the group treated with 100 microg V/mL was observed. The glucose tolerance in the vanadate-treated mice with 10 and 100 microg V/mL was remarkably improved compared with the control group. Biochemical analyses at the end of experiments demonstrated that a distinct tendency for the glucose and hemoglobin A1c (HbA1c) levels to decrease with vanadate treatment in the blood was also observed. The glutamic pyruvic transaminase, glutamic oxaloacetate transaminase, blood urea nitrogen, triglyceride, high-density lipoprotein, and total cholesterol levels in plasma were lower in the higher vanadium groups than those in the control group. These results indicate that vanadium effectively produced the glucose-lowering effect at a higher dose than that at a low dose of vanadium in drinking water, without any overt signs of toxicity.

Animals↗
PubMed2001

Toxic effects of six plant oils alone and in combination with controlled atmosphere on Liposcelis bostrychophila (Psocoptera: Liposcelididae).

Six plant essential oils alone as repellent and fumigant, and in combination with the controlled atmosphere against Liposcelis bostrychophila Badonnel were assessed in the laboratory. These essential oils were extracted from the leaves of six source plants: Citrus tangerina Tanaka, Citrus aurantium L., Citrus bergamia Risso et Poiteau, Pinus sylvestris L., Cupressus funebris End]., and Eucalyptus citriodora Hook. The repellency test indicated that L. bostrychophila adults were repelled by filter paper strips treated with six essential oils. Of these essential oils, the C. funebris oil was most effective followed by that of F. sylvestris, C. tangerina, C. bergamia, and E. citriodora. The average repellency of the C. aurantium oil against L. bostrychophila adults was significantly lower than other five test oils by day 14. These essential oils had a high level of toxicity in the fumigation assay against L. bostrychophila adults at both 10 and 20 ppm. When combined with two controlled atmosphere treatments (12% CO2 + 9% O2, and 10% CO2 + 5% O2, balanced N2), the toxicity of plant oils was enhanced significantly.

Animals↗
PubMed2001

[Studies on the chemical constituents of the seeds from Artabostrys hexapetalus (Annonaceae)].

AIM: To study the chemical constituents of the seeds from Artabotrys hexapetalus (L.f.) Bhandari (Annonaceae). METHODS: Various chromatographic techniques were used to separate and purify the constituents. Their structures were elucidated on the physico-chemical properties and spectral data. RESULTS: Eight compounds were isolated from the seeds of A. hexapetalus. They were identified as four neolignans: isoamericanin A (1), isoamericanol A (2), americanin B (3) and artabotrycinol (4), a semiterpenoid: (R)-artabotriol (5) and others: palmitic acid (6), beta-sitosterol (7) and daucosterol (8). CONCLUSION: Artabotrycinol (4) and (R)-artabotriol (5) are new compounds. Three other neolignans were isolated from this plant for the first time.

Annonaceae↗
PubMed2000

In vitro and in vivo functional analysis of human T cell lymphotropic virus type 1 pX open reading frames I and II.

Human T lymphotropic virus type 1 (HTLV-1) is a complex retrovirus containing regulatory and accessory genes encoded in four open reading frames (ORF I-IV) of the pX region. It is not clear what role pX ORFs I and II-encoded proteins have in the pathogenesis of the lymphoproliferative diseases associated with HTLV-1 infection. The conserved ORF I encodes for a hydrophobic 12-kDa protein, p12, (I) that contains four SH3 binding motifs (PXXP) that localizes to cellular endomembranes when overexpressed in cultured cells. Differential splicing of pX ORF II results in the production of two nuclear proteins, p13(II) and p30(II). p13(II) also localizes to mitochondria. p30(II) shares homology with the POU family of transcription factors. We have identified functional roles of pX ORF I and ORF II in establishment and maintenance of infection in a rabbit model. To functionally study p12(I) we have tested a proviral clone with selective ablation of ORF I (ACH.p12(I)) for its ability to infect quiescent peripheral blood mononuclear cells (PBMC). Our data indicate that T cells infected with the wild-type clone of HTLV-1 (ACH) are more efficient than ACH.p12(I) in infecting quiescent PBMC. These findings parallel our animal model data and suggest a role for p12(I) in the activation of quiescent lymphocytes, a prerequisite for effective viral replication in vivo. To test the ability of p30(II) to function as a transcription factor we have constructed p30(II) as a Gal4-fusion protein. When transfected with Gal4-driven luciferase reporter genes, the p30(II)-Gal4-fusion protein induces transcriptional activity up to 50-fold in both 293 and HeLa-Tat cells. These systems will be useful to identify molecular mechanisms that explain the functional role of pX ORF I and ORF II-encoded proteins in HTLV-1 replication.

Animals↗
PubMed2000

Independent adaptation to riverine habitats allowed survival of ancient cetacean lineages.

The four species of "river dolphins" are associated with six separate great river systems on three subcontinents and have been grouped for more than a century into a single taxon based on their similar appearance. However, several morphologists recently questioned the monophyly of that group. By using phylogenetic analyses of nucleotide sequences from three mitochondrial and two nuclear genes, we demonstrate with statistical significance that extant river dolphins are not monophyletic and suggest that they are relict species whose adaptation to riverine habitats incidentally insured their survival against major environmental changes in the marine ecosystem or the emergence of Delphinidae.

Adaptation, Physiological↗
PubMed2000

Structural identification of a key protective B-cell epitope in Lyme disease antigen OspA.

Outer surface protein A (OspA) is a major lipoprotein of the Borrelia burgdorferi spirochete, the causative agent of Lyme disease. Vaccination with OspA generates an immune response that can prevent bacterial transmission to a mammalian host during the attachment of an infected tick. However, the protective capacity of immune sera cannot be predicted by measuring total anti-OspA antibody. The murine monoclonal antibody LA-2 defines an important protective B-cell epitope of OspA against which protective sera have strong levels of reactivity. We have now mapped the LA-2 epitope of OspA using both NMR chemical-shift perturbation measurements in solution and X-ray crystal structure determination. LA-2 recognizes the three surface-exposed loops of the C-terminal domain of OspA that are on the tip of the elongated molecule most distant from the lipid-modified N terminus. The structure suggests that the natural variation at OspA sequence position 208 in the first loop is a major limiting factor for antibody cross-reactivity between different Lyme disease-causing Borrelia strains. The unusual Fab-dominated lattice of the crystal also permits a rare view of antigen flexibility within an antigen:antibody complex. These results provide a rationale for improvements in OspA-based vaccines and suggest possible designs for more direct tests of antibody protective levels in vaccinated individuals.

Amino Acid Sequence↗
PubMed2000

Synaptotagmin regulation of coated pit assembly.

Synaptotagmins bind clathrin AP-2 with high affinity via their second C(2) domain, which indicates they are involved in coated pit function. We now report that expression of synaptotagmins lacking either the second C(2) domain or the entire cytoplasmic region potently inhibit endocytosis. Inhibition was dependent on two intramembrane cysteine residues that were found to be essential for synaptotagmin oligomerization. Cells expressing the wild-type, but not the mutant, truncated synaptotagmin fragment had a reduced number of clathrin-coated pits. These results suggest that the formation of synaptotagmin multimers is an important step in the regulation of coated pit assembly.

Adaptor Protein Complex alpha Subunits↗
PubMed2000

Stress-activated protein kinase-dependent induction of c-fos by Cd(2+) is mediated by MKK7.

Exposure of mesangial cells to ionic Cd(2+) induces the proto-oncogene c-fos, while activating both Erk and stress-activated protein kinase (SAPK) MAP kinase pathways. While we have previously used a pharmacological inhibitor of Erk activation to implicate involvement of this pathway in the induction of c-fos by Cd(2+), the consequences of SAPK activation remained unknown. Here we use dominant negative inhibitors of the SAPK kinases, SEK1 and MKK7, to show that Cd(2+) activates SAPK through MKK7, but that partial inhibition of SAPK alone is insufficient to significantly affect the magnitude of the Cd(2+)-dependent increase in c-fos mRNA. However, inhibition of Erk and SAPK pathways together abrogates the increase, suggesting that these pathways act in concert in the induction of c-fos by this toxic metal.

Animals↗
PubMed2000

The SCAN domain mediates selective oligomerization.

The SCAN domain is described as a highly conserved, leucine-rich motif of approximately 60 amino acids found at the amino-terminal end of zinc finger transcription factors. Although no specific biological function has been attributed to the SCAN domain, its predicted amphipathic secondary structure led to the suggestion that this domain may mediate protein-protein associations. A yeast two-hybrid screen identified members of two SCAN domain protein families that interact with the SCAN domain of the zinc finger protein ZNF202. The interacting ZNF191 protein represents the family of SCAN domain-containing zinc finger proteins, whereas the novel SDP1 protein establishes a new family of genes that encode an isolated SCAN domain. Isolated SCAN domain proteins may form asymmetric homodimers in solution. Biochemical binding studies confirmed the associations of ZNF191 and SDP1 with ZNF202 and established the SCAN domain as a selective hetero- and homotypic oligomerization domain. SCAN mediated protein associations might therefore represent a new regulatory mechanism of transcriptional activity.

Amino Acid Sequence↗
PubMed2000

Separation of anions by ion chromatography-capillary electrophoresis.

Capillary electrophoresis (CE) with a water-soluble ion-exchange polymer in the background electrolyte is very efficient for the separation of organic and inorganic anions because the ion-exchange selectivity, as well as differences in electrophoretic mobility, can be used for separating sample ions. Poly(diallyldimethylammonium chloride) (PDDAC) was employed for this purpose. A very stable electroosmotic flow was obtained between pH 2.3 and 8.5 due to the strong adsorption of PDDAC onto the capillary wall. The effect of ion exchange on the migration of sample anions and their separation was controlled by varying the concentration of PDDAC, the concentration and the type of salt used in the CE background electrolyte. Addition of organic solvent (e.g., acetonitrile) could also modify the sample migration and the separation. Baseline separations were obtained for anions with very similar mobilities, such as bromide and iodide, naphthalenesulfonates, and bi- and tricarboxylic acids. Typical separation efficiencies were between 195,000 and 429,000 theoretical plates per meter. Ten replicate separations gave an average RSD of 1.0% for migration times of the sample anions studied. Excellent separations were obtained for a variety of samples, including a separation of 17 inorganic and organic anions in less than 6 min.

Anions↗
PubMed2000

Evidence of linkage of familial hypoalphalipoproteinemia to a novel locus on chromosome 11q23.

Coronary heart disease (CHD) accounts for half of the 1 million deaths annually ascribed to cardiovascular disease and for almost all of the 1.5 million acute myocardial infarctions. Within families affected by early and apparently heritable CHD, dyslipidemias have a much higher prevalence than in the general population; 20%-30% of early familial CHD has been ascribed to primary hypoalphalipoproteinemia (low HDL-C). This study assesses the evidence for linkage of low HDL-C to chromosomal region 11q23 in 105 large Utah pedigrees ascertained with closely related clusters of early CHD and expanded on the basis of dyslipidemia. Linkage analysis was performed by use of 22 STRP markers in a 55-cM region of chromosome 11. Two-point analysis based on a general, dominant-phenotype model yielded LODs of 2.9 for full pedigrees and 3.5 for 167 four-generation split pedigrees. To define a localization region, model optimization was performed using the heterogeneity, multipoint LOD score (mpHLOD). This linkage defines a region on 11q23.3 that is approximately 10 cM distal to-and apparently distinct from-the ApoAI/CIII/AIV gene cluster and thus represents a putative novel localization for the low HDL-C phenotype.

Cholesterol, HDL↗
PubMed2000

Iodine-129 in human thyroids and seaweed in China.

The concentrations of 129I and the ratios of 129I/127I in normal human thyroids collected in Tianjin, China, and some seaweed samples from the Chinese coast were determined by neutron activation analysis. The mean 129I/127I ratio in these thyroids was found to be 1.13 x 10(-9), which is two orders of magnitude higher than the level of the pre-nuclear era, but one order of magnitude lower than the level in Europe in the post-nuclear era. There is no significant difference between the ratio of 129I/127I in the thyroids for the post-nuclear era from China and other areas, which are considered not to have been directly exposed to 129I emission from a nuclear source, such as Chile, Taiwan and Tokyo. The mean 129I/127I ratio in seaweed from the Chinese coast is 2.35 x 10(-10), approximately two orders of magnitude higher than in seaweed collected in the pre-nuclear age, and similar to that from locations without direct exposure to the emission from nuclear installations, influenced only by global fallout. This indicates that the 129I level in China is within the global fallout background level.

Adult↗
PubMed2000

Activation of parallel mitogen-activated protein kinase cascades and induction of c-fos by cadmium.

Cadmium is a toxic divalent cation that can initiate either mitogenic signals or apoptosis, possibly as a consequence of inducing different patterns of oncogene expression in different cells. We previously showed that Cd(2+) caused transcriptional activation of the c-fos protooncogene in mesangial cells (Wang and Templeton, J. Biol. Chem. 273, 73-79, 1998). The present study was undertaken to identify the signaling pathways that might be involved. Exposure to 10 microM CdCl(2) for 8 h caused a prolonged activation of Erk kinase and accumulation of c-fos mRNA. Inhibition of Erk activation with PD98059 only partially inhibited c-fos induction, indicating that additional pathways are involved. The c-Jun kinase/stress-activated protein kinase (SAPK) was also activated by Cd(2+). All three signals, i.e., Erk activity, SAPK activity, and c-fos mRNA levels in response to Cd(2+) showed a similar biphasic time course with an initial increase at 15-30 min and then a larger and more prolonged increase several hours later. Each signal also showed a similar concentration dependence, with less than 1 microM Cd(2+) causing the initial increase but values above 3 microM being required for the prolonged phase. These events showed high specificity for Cd(2+); other divalent metals tested under the same conditions (Mg(2+), Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), and Hg(2+)) were without significant effects. We conclude that Cd(2+) is a specific inducer of c-fos in mesangial cells, probably through activation of both Erk kinase and SAPK pathways. The similar time and concentration dependence of the response of both pathways to Cd(2+) suggests a common basis for activation.

Animals↗
PubMed2000

Tumor antigen presentation by dermal antigen-presenting cells.

Several phenotypes of antigen-presenting cells are present in the dermis, where they presumably function to present encountered antigens for immune responses. This study examined the ability of dermal antigen-presenting cells to present tumor-associated antigens for the induction of in vivo antitumor immunity. Total murine dermal cells were exposed either to medium alone or to medium containing tumor-associated antigens from S1509a tumor cells. Subsequently, dermal cells were injected subcutaneously at weekly intervals into naïve mice for a total of three immunizations. One week following the final immunization, mice were challenged with living tumor cells. In these experiments, dermal cells pulsed with tumor-associated antigens induced protective immunity to tumor growth. Dermal cells exposed to tumor-associated antigens were also able to elicit delayed-type hypersensitivity after footpad injection into mice previously immunized against S1509a tumor cells. The ability to present tumor-associated antigens for both induction of antitumor immunity and elicitation of delayed-type hypersensitivity was dependent on I-A+ cells and was genetically restricted. Finally, dermal cells tended towards eliciting a greater antitumor delayed-type hypersensitivity response than epidermal cells. These results show that the murine dermis contains antigen-presenting cells capable of processing S1509a tumor antigens for the generation of protective antitumor immunity in vivo.

Animals↗
PubMed2000

Induction of anti-tumor immunity with epidermal cells pulsed with tumor-derived RNA or intradermal administration of RNA.

The skin is well-suited to serve as a substrate for vaccination strategies. In this regard, epidermal cells exposed to granulocyte-macrophage colony-stimulating factor can, upon subcutaneous injection into naïve mice, present a soluble extract of tumor as a source of tumor-associated antigens for the induction of in vivo anti-tumor immunity. Use of RNA for immunization has a potential advantage over this technique. Because RNA can be amplified, only a small amount of tumor is needed for antigen preparation and, as with a soluble extract, it is not necessary to know the molecular nature of the antigen(s) relevant to immunity. To test the hypothesis that RNA-pulsed epidermal cells can induce anti-tumor immunity, total cellular RNA was isolated from the S1509a spindle cell tumor and used to pulse CAF1 epidermal cells enriched for Langerhans cell content and pre-exposed to granulocyte-macrophage colony-stimulating factor. These cells were then injected subcutaneously into naïve CAF1 mice three times at weekly intervals followed by challenge with living S1509a cells. Tumor growth was significantly less than in control animals immunized in an identical fashion but with irrelevant RNA. Digestion of S1509a RNA with RNase prior to pulsing of epidermal cells prevented the development of immunity. In separate experiments, intradermal injection of S1509a RNA into naïve mice three times at weekly intervals also induced immunity to challenge with the tumor. Digestion of S1509a RNA with RNase also prevented development of immunity in this system. Effective anti-tumor immunity can be induced in mice utilizing RNA-pulsed epidermal cells for in vivo immunization or by injecting RNA intradermally into naïve mice.

Animals↗