Biomedical subjects

C Chang

Publications and source records attributed to C Chang.

At least 127 records · Page 7Linked to original sources

PubMed1999

Identification of a virulence-associated protein homolog gene and ISRa1 in a plasmid of Riemerella anatipestifer.

Riemerella anatipestifer is the causative agent of polyserositis of ducks and geese. We have previously reported that a 3.9-kb plasmid, pCFC1, carries protein genes (vapD1 and vapD2) that are similar to virulence-associated genes of other bacteria. In the present study, we report the complete sequence of a second plasmid of 5.6 kb, pCFC2. pCFC2 has a 28% G-C content and three large open reading frames (ORFs). One of the ORFs (designated asVapD1) encodes a polypeptide that shares 53.9, 53.9, 48.3, 48.3 and 46.1% identity with virulence-associated proteins of Dichelobacter nodosus, Actinobacillus actinomycetemcomitans, Neisseria gonorrhoeae, Helicobacter pylori and Haemophilus influenzae, respectively. The second ORF encodes a putative DNA replication protein (RepA3) with 309 amino acids and a molecular mass of approximately 36 kDa. A novel insertion sequence (IS) element, designated ISRa1, was found on the plasmid pCFC2. ISRa1 was flanked by 15-bp imperfect inverted repeats (only one mismatched nucleotide). ISRa1 contained an ORF encoding a putative transposase of 292 amino acids. Southern blot analysis indicated that in R. anatipestifer strains examined, ISRa1 was present with 2-20 copies (at least). ISRa1 displayed a sequence approximately 35% homologous to the putative IS982 and RSBst-alpha from Lactococcus lactis ssp. cremoris SK11 and Bacillus stearothermophilus CU21. Three hybridization patterns of genomic DNA of eight R. anatipestifer strains with an ISRa1 probe indicated that ISRa1 might be a useful tool for epidemiological studies.

Amino Acid Sequence↗

PubMed1999

Suppression of Delta(5)-androstenediol-induced androgen receptor transactivation by selective steroids in human prostate cancer cells.

Our earlier report suggested that androst-5-ene-3beta,7beta-diol (Delta(5)-androstenediol or Adiol) is a natural hormone with androgenic activity and that two potent antiandrogens, hydroxyflutamide (Eulexin) and bicalutamide (Casodex), fail to block completely the Adiol-induced androgen receptor (AR) transactivation in prostate cancer cells. Here, we report the development of a reporter assay to screen several selective steroids with anti-Adiol activity. Among 22 derivatives/metabolites of dehydroepiandrosterone, we found 4 steroids [no. 4, 1,3,5(10)-estratriene-17alpha-ethynyl-3, 17beta-diol; no. 6, 17alpha-ethynyl-androstene-diol; no. 8, 3beta, 17beta-dihydroxy-androst-5-ene-16-one; and no. 10, 3beta-methylcarbonate-androst-5-ene-7,17-dione] that have no androgenic activity and could also block the Adiol-induced AR transactivation in prostate cancer PC-3 cells. Interestingly, these compounds, in combination with hydroxyflutamide, further suppressed the Adiol-induced AR transactivation. Reporter assays further showed that these four anti-Adiol steroids have relatively lower glucocorticoid, progesterone, and estrogenic activity. Together, these data suggest some selective steroids might have anti-Adiol activity, which may have potential clinical application in the battle against the androgen-dependent prostate cancer growth.

Androstenediol↗

PubMed1999

Adsorption properties and breakthrough model of 1,1-dichloro-1-fluoroethane on activated carbons.

Hydrochlorofluorocarbons (HCFCs), among the major replacements for chlorofluorocarbons (CFCs) and chlorinated solvents, are now considered to the prime contribution to the stratospheric ozone depletion. This paper reports the adsorption equilibrium of 1,1-dichloro-1-fluoroethane (HCFC-141b) vapor in air streams on the commercial activated carbons PCB and BPL, which were made from coconut shell and bituminous coal, respectively, at 283, 293, 303, and 313 K. The experimental results show that within the experimental conditions, the adsorption capacities of the two adsorbents for HCFC-141b in dry air streams can be well-fitted by the Langmuir, Freundlich, and Dubinin-Radushkevich (D-R) adsorption equations. The physical properties of the adsorbents are consistent with the isotherm parameters obtained from the adsorption results. In addition, available experimental data of adsorption breakthrough at 283 K were used in conjunction with the Yoon and Nelson model to generate theoretical breakthrough curves based on the obtained values of parameters. In each case, calculated theoretical breakthrough curves are in agreement with corresponding experimental data.

Journal Article↗

PubMed1999

Isolation and characterization of ARA160 as the first androgen receptor N-terminal-associated coactivator in human prostate cells.

The androgen receptor (AR) is a member of the steroid receptor superfamily that may require coactivators for proper or maximal transactivation. Using a purified AR N-terminal peptide as a probe to screen the human testis expression library, we identified an androgen-enhanced AR N-terminal-associated protein ARA160, which consists of 1,093 amino acids with an apparent molecular mass of 160 kDa. Sequence comparison in GenBank(TM) reveals that ARA160 shares an identical sequence with a HIV-1 TATA element modulatory factor, TMF. The far-Western blotting and co-immunoprecipitation assays demonstrate that the AR can interact directly with ARA160/TMF. Affinity gel pull-down and mammalian two-hybrid assays further suggest androgen can enhance significantly the interaction between AR and ARA160. Transient transfection assays demonstrated that ARA160 might function as a coactivator for AR-mediated transactivation in human prostate cancer PC-3 cells. Our data further suggest that this AR N-terminal coactivator can function cooperatively with AR C-terminal coactivator, ARA70, in PC-3 cells. Together, our data demonstrate that ARA160 might represent the first identified androgen-enhanced N-terminal coactivator for the AR.

Amino Acid Sequence↗

PubMed1999

The linkage of Kennedy's neuron disease to ARA24, the first identified androgen receptor polyglutamine region-associated coactivator.

Although the linkage of polyglutamine (poly-Q) repeat expansion in the androgen receptor (AR) to Kennedy's disease (X-linked spinal and bulbar muscular atrophy) was a major step forward, the detailed molecular mechanism of how the change in poly-Q length contributes to the disease remains unclear. Here we report the identification of a nuclear G-protein, Ras-related nuclear protein/ARA24, as the first AR coactivator that can bind differentially with different lengths of poly-Q within AR. In the yeast and mammalian reciprocal interacting assays, our data suggested the interaction of AR N-terminal domain with ARA24 diminishes as the poly-Q length increases. The coactivation of ARA24 also diminishes with the poly-Q expansion within AR. Deletion of the acidic hexapeptide (DEDDDL) at the C terminus of ARA24 further enhances its AR coactivation. Together, our data suggest that poor interaction and weaker coactivation of ARA24 to the longer poly-Q AR in the X-linked spinal and bulbar muscular atrophied AR could contribute to the weaker transactivation of AR. The consequence of poor interaction and weak coactivation may eventually lead to the partial androgen insensitivity during the development of Kennedy's disease.

Amino Acid Sequence↗

PubMed1999

Identification of ARA70 as a ligand-enhanced coactivator for the peroxisome proliferator-activated receptor gamma.

In an effort to understand transcriptional regulation by the peroxisome proliferator-activated receptor gamma (PPARgamma), we have investigated its potential interaction with coregulators and have identified ARA70 as a ligand-enhanced coactivator. ARA70 was initially described as a coactivator for the androgen receptor (AR) and is expressed in a range of tissues including adipose tissue (Yeh, S., and Chang, C. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 5517-5521). Here we show that ARA70 and PPARgamma specifically interact by coimmunoprecipitation and in a mammalian two-hybrid assay. PPARgamma and ARA70 interact in the absence of the PPARgamma ligand 15-deoxy-Delta12,14-prostaglandin J2, although the addition of exogenous ligand enhances this interaction. Similarly, in transient transfection of DU145 cells, cotransfection of PPARgamma and ARA70 induces transcription from reporter constructs driven by either three copies of an isolated PPAR response element or the natural promoter of the adipocyte fatty acid-binding protein 2 in the absence of exogenous 15-deoxy-Delta12,14-prostaglandin J2. However, this PPARgamma-ARA70 transactivation is enhanced by the addition of ligand. Thus, ARA70 can function as a ligand-enhanced coactivator of PPARgamma. Finally, we show that AR can squelch PPARgamma-ARA70 transactivation, which suggests that cross-talk may occur between PPARgamma- and AR-mediated responses in adipocytes.

Cell Line↗

PubMed1999

Differential regulation of direct repeat 3 vitamin D3 and direct repeat 4 thyroid hormone signaling pathways by the human TR4 orphan receptor.

In situ hybridization analysis demonstrated that abundant testicular orphan receptor (TR4) transcripts were detected in kidney, intestine, and bone, which are vitamin D3 target organs. Cell transfection studies also demonstrated that the expression of the vitamin D3 target gene, 25-hydroxyvitamin D3 24-hydroxylase, can be repressed by TR4 through high affinity binding (Kd = 1.32 nM) to the direct repeat 3 vitamin D3 receptor response element (DR3VDRE). This TR4-mediated repression of DR3VDRE is in contrast to our earlier report that TR4 could induce thyroid hormone target genes containing a direct repeat 4 thyroid hormone response element (DR4T3RE). Electrophoretic mobility shift assay using several TR4 monoclonal antibodies when combined with either TR4-DR3VDRE or TR4-DR4T3RE showed a distinct supershifted pattern, and proteolytic analysis further demonstrated distinct digested peptides with either TR4-DR3VDRE or TR4-DR4T3RE. These results may therefore suggest that TR4 can adapt to different conformations once bound to DR3VDRE or DR4T3RE. The consequence of these different conformations of TR4-DR3VDRE and TR4-DR4T3RE may allow each of them to recruit different coregulators. The differential repression of TR4-mediated DR3VDRE and DR4T3RE transactivation by the receptor interacting protein 140, a TR4 coregulator, further strengthens our hypothesis that the specificity of gene regulation by TR4 can be modulated by protein-DNA and protein-protein interactions.

Animals↗

PubMed1999

Isolation of Ku70-binding proteins (KUBs).

DNA-dependent protein kinase (DNA-PK) plays a critical role in resealing DNA double-stand breaks by non-homologous end joining. Aside from DNA-PK, XRCC4 and DNA ligase IV, other proteins which play a role(s) in this repair pathway remain unknown; DNA-PK contains a catalytic subunit (DNA-PKcs) and a DNA binding subunit (Ku70 and Ku80). We isolated Ku70-binding proteins (KUB1-KUB4) using yeast two-hybrid analyses. Sequence analyses revealed KUB1 to be apolipoprotein J (apoJ), also known as X-ray-inducible transcript 8 (XIP8), testosterone-repressed prostate message-2 (TRPM-2) and clusterin. KUB2 is Ku80. KUB3 and KUB4 are unknown, >10 kb trans-cripts. Interactions of apoJ/XIP8 or KUB3 with Ku70 were confirmed by co-immunoprecipitation analyses in MCF-7:WS8 breast cancer or IMR-90 normal lung fibroblast cells, respectively. The interaction of apoJ/XIP8 with Ku70 was confirmed by far-western analyses. Stable over-expression of full-length apoJ/XIP8 in MCF-7:WS8 caused decreased Ku70/Ku80 DNA end binding that was restored by apoJ/XIP8 monoclonal antibodies. The role of apoJ/XIP8 in ionizing radiation resistance/sensitivity is under investigation.

Animals↗

PubMed1999

Crystal structures of thermostable xylose isomerases from Thermus caldophilus and Thermus thermophilus: possible structural determinants of thermostability.

The crystal structures of highly thermostable xylose isomerases from Thermus thermophilus (TthXI) and Thermus caldophilus (TcaXI), both with the optimum reaction temperature of 90 degrees C, have been determined by X-ray crystallography. The model of TcaXI has been refined to an R-factor of 17.8 % for data extending to 2.3 A and that of TthXI to 17.1 % for data extending to 2.2 A. The tetrameric arrangement of subunits characterized by the 222-symmetry and the tertiary fold of each subunit in both TcaXI and TthXI are basically the same as in other xylose isomerases. Each monomer is composed of two domains. Domain I (residues 1 to 321) folds into the (beta/alpha)8-barrel. Domain II (residues 322 to 387), lacking beta-strands, makes extensive contacts with domain I of an adjacent subunit. Each monomer of TcaXI contains ten beta-strands, 15 alpha-helices, and six 310-helices, while that of TthXI contains ten beta-strands, 16 alpha-helices, and five 310-helices. Although the electron density does not indicate the presence of bound metal ions in the present models of both TcaXI and TthXI, the active site residues show the conserved structural features. In order to understand the structural basis for thermostability of these enzymes, their structures have been compared with less thermostable XIs from Arthrobacter B3728 and Actinoplanes missouriensis (AXI and AmiXI), with the optimum reaction temperatures of 80 degrees C and 75 degrees C, respectively. Analyses of various factors that may affect protein thermostability indicate that the possible structural determinants of the enhanced thermostability of TcaXI/TthXI over AXI/AmiXI are (i) an increase in ion pairs and ion-pair networks, (ii) a decrease in the large inter-subunit cavities, (iii) a removal of potential deamidation/isoaspartate formation sites, and (iv) a shortened loop.

Aldose-Ketose Isomerases↗

PubMed1999

From HER2/Neu signal cascade to androgen receptor and its coactivators: a novel pathway by induction of androgen target genes through MAP kinase in prostate cancer cells.

Overexpression of the HER2/Neu protooncogene has been linked to the progression of breast cancer. Here we demonstrate that the growth of prostate cancer LNCaP cells can also be increased by the stable transfection of HER2/Neu. Using AG879, a HER2/Neu inhibitor, and PD98059, a MAP kinase inhibitor, as well as MAP kinase phosphatase-1 (MPK-1), in the transfection assay, we found that HER2/Neu could induce prostate-specific antigen (PSA), a marker for the progression of prostate cancer, through the MAP kinase pathway at a low androgen level. Reporter assays and mammalian two-hybrid assays further suggest this HER2/Neu-induced androgen receptor (AR) transactivation may function through the promotion of interaction between AR and AR coactivators, such as ARA70. Furthermore, we found this HER2/Neu --> MAP kinase --> AR-ARAs --> PSA pathway could not be blocked completely by hydroxyflutamide, an antiandrogen used in the treatment of prostate cancer. Together, these data provide a novel pathway from HER2/Neu to AR transactivation, and they may represent one of the reasons for the PSA re-elevation and hormone resistance during androgen ablation therapy in prostate cancer patients.

Androgen Antagonists↗

PubMed1999

Cloning and characterization of human prostate coactivator ARA54, a novel protein that associates with the androgen receptor.

Androgen receptor (AR) is a member of the steroid receptor superfamily that may require coactivators for proper or maximal transactivation. Using a yeast two-hybrid screening followed by mammalian cell analyses, we identified a novel ligand-dependent AR-associated protein, ARA54, which consists of 474 amino acids with a molecular mass of 54 kDa. We demonstrated that ARA54 might function as a preferential coactivator for AR-mediated transactivation in human prostate cancer DU145 cells. Interestingly, our data also showed that ARA54 could significantly enhance the transcriptional activity of LNCaP mutant AR (ARt877a) but not wild type AR or another mutant AR (ARe708k) in the presence of 10 nM 17beta-estradiol or 1 microM hydroxyflutamide. These results imply that both ARA54 and the positions of the AR mutation (877 versus 708) might contribute to the specificity of AR-mediated transactivation. Our findings further demonstrated that the C-terminal domain of ARA54 can serve as a dominant negative inhibitor and exogenous full-length ARA54 can reverse this squelching effect on AR transcriptional activity. Co-expression of ARA54 with other AR coactivators, such as ARA70 or SRC-1, showed additive stimulation of AR-mediated transactivation, which indicates that these cofactors may function individually as AR coactivators to induce AR target gene expression. Through our findings, we have identified and characterized a novel AR coactivator, ARA54, which may play an important role in the AR signaling pathway in human prostate.

Amino Acid Sequence↗

PubMed1999

Cloning and characterization of androgen receptor coactivator, ARA55, in human prostate.

Androgen receptor (AR) is a hormone-activated transcriptional factor that can bind to androgen response elements and that regulates the transcription of target genes via a mechanism that presumably involves cofactors. We report here the cloning of a novel AR coactivator ARA55 using a yeast two-hybrid system. ARA55 consists of 444 amino acids with the predicted molecular mass of 55 kDa and its sequence shows very high homology to mouse hic5, a TGF-beta1-inducible gene. Yeast and mammalian two-hybrid systems and co-immunoprecipitation assays all prove ARA55 can bind to AR in a ligand-dependent manner. Transient transfection assay in prostate cancer DU145 cells further demonstrates that ARA55 can enhance AR transcriptional activity in the presence of 1 nM dihydrotestosterone or its antagonists such as 100 nM 17beta-estradiol or 1 microM hydroxyflutamide. Our data also suggest the C-terminal half of ARA55, which includes three LIM motifs, is sufficient to interact with AR. Northern blot and polymerase chain reaction quantitation showed ARA55 can be expressed differently in normal prostate and prostate tumor cells. Together, our data suggests that ARA55 may play very important roles in the progression of prostate cancer by the modulation of AR transactivation.

Amino Acid Sequence↗

PubMed1999

Reciprocal EGF signaling back to the uterus from the induced C. elegans vulva coordinates morphogenesis of epithelia.

BACKGROUND: Reciprocal signaling between distinct tissues is a general feature of organogenesis. Despite the identification of developmental processes in which coordination requires reciprocal signaling, little is known regarding the underlying molecular details. Here, we use the development of the uterine-vulval connection in the nematode Caenorhabditis elegans as a model system to study reciprocal signaling. RESULTS: In C. elegans, development of the uterine-vulval connection requires the specification of uterine uv1 cells and morphogenesis of 1 degrees -derived vulval cells. LIN-3, an epidermal growth factor (EGF) family protein, is first produced by the gonadal anchor cell to induce vulval precursor cells to generate vulval tissue. We have shown that lin-3 is also expressed in the 1 degrees vulval lineage after vulval induction and that the 1 degrees vulva is necessary to induce the uv1 uterine cell fate. Using genetic and cell biological analyses, we found that the specification of uterine uv1 cells is dependent on EGF signaling from cells of the 1 degrees vulval lineages to a subset of ventral uterine cells of the gonad. RAS and RAF are necessary for this signaling. We also found that EGL-38, a member of the PAX family of proteins, is necessary for transcription of lin-3 in the vulva but not in the anchor cell. A let-23 mutation that confers ligand-independent activity bypasses the requirement for EGL-38 in specification of the uv1 cell fate. CONCLUSIONS: We have shown how relatively simple EGF signals can be used reciprocally to specify the uterine-vulval connection during C. elegans development.

Animals↗

PubMed1999

A thermostable xylose isomerase from Thermus caldophilus: biochemical characterization, crystallization and preliminary X-ray analysis.

A highly thermostable xylose isomerase from Thermus caldophilus has been expressed in Escherichia coli. The purified enzyme has an optimum temperature of 363 K. It has been crystallized at room temperature using ammonium sulfate as a precipitant. The crystal belongs to the orthorhombic space group P212121, with unit-cell parameters a = 84.35, b = 123.60, c = 140.24 A. The presence of one molecule of tetrameric xylose isomerase in the asymmetric unit gives a crystal volume per protein mass (Vm) of 2.1 A3 Da-1 and a solvent content of 41% by volume. The crystals initially showed diffraction to 1.7 A Bragg spacing with synchrotron X-rays, and a set of native data extending to 2.3 A resolution has been collected.

Aldose-Ketose Isomerases↗

PubMed1999

Temporal evolution of 3-nitropropionic acid-induced neurodegeneration in the rat brain by T2-weighted, diffusion-weighted, and perfusion magnetic resonance imaging.

An appropriate detecting technique is necessary for the early detection of neurodegenerative diseases. 3-Nitropropionic acid-intoxicated rats serve as the animal model for one neurodegenerative disease, Huntington's disease. Non-invasive diffusion- and T2-weighted magnetic resonance imaging were applied to study temporal evolution and spatial distribution of brain lesions which were produced by intravenous injection of 3-nitropropionic acid in rats. Lesions in the striatum, hippocampus, and corpus callosum but not in the cortex were observed 3 and 4.5 h after 3-nitropropionic acid injection (30 mg/kg) on the diffusion- and T2-weighted images, respectively (n = 6). The results demonstrated that the diffusion-weighted imaging is not only superior to T2-weighted imaging in detecting onset of 3-nitropropionic acid-induced excitotoxic brain damage but also differentiates lesion and non-lesion areas with better spatial resolution than T2-weighted imaging. Additionally, to correlate structural alterations with pathophysiological conditions, dynamic susceptibility contrast magnetic resonance imaging was performed before and 4 h after 3-nitropropionic acid administration (n = 8). The relative cerebral blood volume was significantly elevated in the striatum (P < 0.001) but not in the cortex after 3-nitropropionic acid administration. The changes in regional relative cerebral blood volume were well correlated to the changes in signal intensities in the corresponding areas on the diffusion- and T2-weighted images. The combined structural and functional information in this study may provide new insights and therapeutic strategies in treating neurodegenerative diseases.

Animals↗

PubMed1999

The ethylene-response pathway: signal perception to gene regulation.

Tremendous strides have been made in the past year toward elucidating the ethylene-response pathway. Ethylene is perceived by a family of histidine kinase-like receptors, which negatively regulate ethylene responses. Binding of ethylene requires a copper cofactor, and proper receptor function relies on a copper transporter. Downstream, EIN2 is a structurally novel protein containing an integral membrane domain. In the nucleus, the EIN3 family of DNA-binding proteins regulates transcription in response to ethylene, and an immediate target of EIN3 is a DNA-binding protein of the AP2/EREBP family.

Arabidopsis↗

PubMed1999

C. elegans vulval development as a model system to study the cancer biology of EGFR signaling.

Molecular genetic studies of C. elegans vulval development have helped to define an evolutionarily conserved signaling pathway from an EGF-like ligand through EGF-receptor, Ras and MAP kinase to the nucleus. Further studies have identified novel positive regulators such as KSR-1 and SUR-8/SOC-2 and negative regulators such as cbl/SLI-1. The many negative regulatory proteins might serve to prevent inappropriate signaling, and thus are analogous to tumor suppressor genes.

Animals↗

PubMed1999

Identification of the histamine H1 receptor gene as a differentially repressed target of the human TR2 orphan receptor.

We have identified a DNA response element (TR2RE-HR) in the 3' flanking region of the human histamine H1 receptor gene as a target for the TR2 orphan receptor, a member of the steroid/thyroid hormone receptor superfamily. The application of both tetracycline inducible and improved differential display systems has allowed us to isolate a cDNA fragment differentially regulated by the expression of the TR2 orphan receptor. Northern blot and sequencing analysis demonstrated that the expression of the human histamine H1 receptor gene was differentially repressed by the TR2 orphan receptor. Electrophoretic mobility shift assay further revealed a specific binding (dissociation constant = 26.2 nM) between the TR2 orphan receptor and the wildtype TR2RE-HR, but not the mutant TR2RE-HR. In addition, reporter gene expression assay indicated that the TR2 orphan receptor may suppress the expression of luciferase activities in a dose-dependent manner via the TR2RE-HR in HeLa cells. Our results demonstrate that the histamine H1 receptor gene could represent one of the target genes directly regulated by the human TR2 orphan receptor.

Base Sequence↗