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C Wu

Publications and source records attributed to C Wu.

At least 433 records · Page 24Linked to original sources
PubMed1995

Displacement of sequence-specific transcription factors from mitotic chromatin.

The general inhibition in transcriptional activity during mitosis abolishes the stress-inducible expression of the human hsp70 gene. Among the four transcription factors that bind to the human hsp70 promoter, the DNA-binding activities of three (C/EBP, GBP, and HSF1) were normal, while Sp1 showed reduced binding activity in mitotic cell extracts. In vivo footprinting and immunocytochemical analyses revealed that all of the sequence-specific transcription factors were displaced from promoter sequences as well as from bulk chromatin during mitosis. The correlation of transcription factor displacement with chromatin condensation suggests an involvement of chromatin structure in mitotic repression. However, retention of DNase I hypersensitivity suggests that the hsp70 promoter was not organized in a canonical nucleosome structure in mitotic chromatin. Displacement of transcription factors from mitotic chromosomes could present another window in the cell cycle for resetting transcriptional programs.

Base Sequence↗
PubMed1995

Polymorphisms and probable lack of mutation in a human mutT homolog, hMTH1, in hereditary nonpoliposis colorectal cancer.

The human MTH1 gene, a homolog of the E. coli mutator gene mutT, encodes 8-oxo-dGTPase that degrades a mutagenic substrate for DNA synthesis. To determine whether this gene is associated with hereditary nonpolyposis colorectal cancer (HNPCC), we examined the hMTH1 sequence in 32 HNPCC cases by means of polymerase chain reaction-single strand conformation polymorphism and sequencing analyses. Three different DNA variants were identified in normal and the corresponding tumor DNA. The first variant, a G to A transition at codon 83, changes valine to methionine and was detected in 9 HNPCC cases. The same change was detected in 5 of 30 unrelated healthy individuals, suggesting that it is not associated with a marked HNPCC predisposition. The second variant is a silent C to T transition at codon 119. Another C to T transition at 31 bp upstream of the beginning of exon 4 was also found. However, specific mutations in hMTH1 were detected in neither normal nor tumor cells from HNPCC patients. These results indicate that hMTH1 is not directly involved in HNPCC.

Adaptor Proteins, Signal Transducing↗
PubMed1995

Association of the yeast pheromone response G protein beta gamma subunits with the MAP kinase scaffold Ste5p.

The mating response pathway of the yeast Saccharomyces cerevisiae includes a heterotrimeric guanine nucleotide-binding protein (G protein) that activates a mitogen-activated protein MAP kinase cascade by an unknown mechanism. An amino-terminal fragment of the MAP kinase scaffold protein Ste5p that interfered with pheromone-induced cell cycle arrest was identified. A haploid-specific interaction between the amino terminus of Ste5p and the G protein beta subunit Ste4p was also detected in a two-hybrid assay, and the product of a signaling-defective allele of STE4 was defective in this interaction. In cells with a constitutively activated pheromone response pathway, epitope-tagged Ste4p was coimmunoprecipitated with Ste5p. Thus, association of the G protein and the MAP kinase cassette via the scaffolding protein Ste5p may transmit the G protein signal.

Adaptor Proteins, Signal Transducing↗
PubMed1995

Arabidopsis heat shock factor is constitutively active in Drosophila and human cells.

Heat shock factors (HSF) are the transcriptional activators of the heat shock response. The conversion of constitutively expressed HSF to a form that can bind DNA requires the trimerization of the protein, involving leucine zipper interactions as shown for yeast, Drosophila, chicken and human HSFs. Like other metazoan HSFs, the endogenous Arabidopsis HSF displays heat shock-inducible DNA-binding activity in gel retardation assays. The heat shock-inducible binding of a recombinant Arabidopsis HSF (ATHSF1) expressed in Arabidopsis plants suggests that ATHSF1 is the major heat shock factor regulating the heat stress response. However, on transient expression in Drosophila and human cells, ATHSF1 fails to exhibit proper regulation, as demonstrated by constitutive binding to DNA, and by constitutive expression of a chloramphenicol acetyltransferase (CAT) reporter gene under the control of the Drosophila hsp70 promoter. These results suggest that the regulation of ATHSF1 is normally dependent on a specific factor that inhibits the DNA-binding and transcriptional activities under non-heat shock conditions.

Animals↗
PubMed1995

Molecular characterization of Ste20p, a potential mitogen-activated protein or extracellular signal-regulated kinase kinase (MEK) kinase kinase from Saccharomyces cerevisiae.

The Ste20p protein kinase was immunopurified from yeast cells and analyzed in an in vitro assay system. Ste20p immune complexes exhibited autophosphorylating activity at serine and threonine residues and specifically phosphorylated a bacterially expressed glutathione S-transferase (GST) fusion of Ste11p (a mitogen-activated protein or extracellular signal-regulated kinase kinase (MEK) kinase homologue) at serine and threonine residues. In contrast, GST fusions either of Ste7p (a MEK homologue) or the beta-subunit of the mating response G-protein and immunoprecipitated Ste5p were not phosphorylated by the Ste20p immune complexes. Myelin basic protein was identified as an excellent in vitro substrate, whereas histone H1 was only poorly phosphorylated. Evidence was obtained that autophosphorylation might play a regulatory role for the in vitro kinase activity. The in vitro activity was found to be Ca(2+)-independent. Both the in vivo and in vitro activities were abolished by mutational changes of either the conserved lysine residue 649 within the ATP binding site or threonine 777 between the catalytic subdomains VII and VIII. Wild-type Ste20p and the catalytically inactive T777A mutant were identified as phosphoproteins in vivo. The phosphorylation occurred at serine and threonine residues independent of pheromone stimulation. Based on the genetically determined significance of Ste20p in pheromone signal transduction and on our in vitro studies, we propose the model that Ste20p represents a yeast MEK kinase kinase whose function is to link G-protein-coupled receptors through G beta gamma to a mitogen-activated protein kinase module.

Amino Acid Sequence↗
PubMed1995

Stimulation of the DNA-dependent protein kinase by RNA polymerase II transcriptional activator proteins.

The DNA-dependent protein kinase (DNA-PK) phosphorylates RNA polymerase II and a number of transcription factors. We now show that the activity of DNA-PK is directly stimulated by certain transcriptional activator proteins, including the human heat shock transcription factor 1 (HSF1) and a transcriptionally active N-terminal 147 amino acid GAL4 derivative. Stimulation of DNA-PK activity required specific sequences in the activator proteins outside the minimal DNA binding domains. The stimulation of DNA-PK activity also required DNA and was greater with DNA containing relevant activator binding sites. Comparison of different HSF binding fragments showed that optimal stimulation occurred when two HSF binding sites were present. Stimulation with HSF and GAL4 was synergistic with Ku protein, another regulator of DNA-PK activity. DNA-PK is tightly associated with the transcriptional template, and an increase in its activity could potentially influence transcription through the phosphorylation of proteins associated with the transcription complex.

Antigens, Nuclear↗
PubMed1995

A bivariate limiting distribution of tumor latency time.

The model of radiation carcinogenesis, proposed earlier by Klebanov, Rachev, and Yakovlev [8] substantiates the employment of limiting forms of the latent time distribution at high dose values. Such distributions arise within the random minima framework, the two-parameter Weibull distribution being a special case. This model, in its present form, does not allow for carcinogenesis at multiple sites. As shown in the present paper, a natural two-dimensional generalization of the model appears in the form of a Weibull-Marshall-Olkin distribution. Similarly, the study of a randomized version of the model based on the negative binomial minima scheme results in a bivariate Pareto-Marshall-Olkin distribution. In the latter case, an estimate for the rate of convergence to the limiting distribution is given.

Animals↗
PubMed1995

Bone marrow purging with dibutyl phthalate--experimental basis and a preliminary clinical application.

It has been found recently that di-n-butyl phthalate (DBP) possesses a new pharmacological activity in the selective elimination of tumor cells from marrow. In combination with long-term liquid culture of marrow cells in vitro, DBP purged autologous bone marrow transplantation has first been applied to a case of acute myeloid leukemia at the first complete remission with successful reconstitution of hematopoiesis.

Adult↗
PubMed1995

Heat shock transcription factors: structure and regulation.

Organisms respond to elevated temperatures and to chemical and physiological stresses by an increase in the synthesis of heat shock proteins. The regulation of heat shock gene expression in eukaryotes is mediated by the conserved heat shock transcription factor (HSF). HSF is present in a latent state under normal conditions; it is activated upon heat stress by induction of trimerization and high-affinity binding to DNA and by exposure of domains for transcriptional activity. Analysis of HSF cDNA clones from many species has defined structural and regulatory regions responsible for the inducible activities. The heat stress signal is thought to be transduced to HSF by changes in the physical environment, in the activity of HSF-modifying enzymes, or by changes in the intracellular level of heat shock proteins.

Amino Acid Sequence↗
PubMed1995

Functional expression of yoked human chorionic gonadotropin in baculovirus-infected insect cells.

hCG is a glycoprotein hormone composed of an alpha-subunit, common to all gonadotropins and to TSH, and a hormone-specific beta-subunit. The non-covalent association of the two subunits is an obligatory step for the formation of biologically active hormones. The correct assembly of the heterodimer is also important for efficient secretion of the hormone, receptor binding, and signal transduction. Herein, we have demonstrated that expression of the two subunits from independent promoters present in a single recombinant baculovirus resulted in subunit association and secretion of biologically active holoprotein by the insect cells. To determine whether the active conformation of heterodimer could be achieved when the two subunits were synthesized in tandem on a single polypeptide chain, two single chain or yoked hCG1, the C-terminus of the complete beta-subunit (145 amino acid residues) was conjoined to the N-terminus of the alpha-subunit. Yoked hCG2 was similar, except that it contained the N-terminal 123 amino acid residues of the beta-subunit. Both yoked hCG molecules bound LH/CG receptor with high affinity and stimulated adenylate cyclase and progesterone levels in transformed mouse Leydig (MA-10) cells. Therefore, the alpha- and beta-subunits are able to fold into a biologically active conformation when covalently linked. Interestingly, when compared with urinary hCG, the hormone expressed in baculovirus-infected insect cells binds to the LH/CG receptor with higher affinity, but exhibits diminished signaling, thus providing another example of a partial dissociation between receptor binding and activation.

Animals↗
PubMed1995

The role of alpha 4 beta 1 integrin in cell motility and fibronectin matrix assembly.

The alpha 4 beta 1 integrin has been suggested to play important roles in embryogenesis and pathogenesis of many diseases which involve both cell adhesion and cell migration. Previous studies using anti-alpha 4 beta 1 antibodies and fibronectin (Fn) fragments have suggested that alpha 4 beta 1 integrins may be involved in cell motility on Fn and vascular cell adhesion molecule-1 (VCAM-1). However, the cells used in these studies also express other Fn integrin receptors including alpha 5 beta 1 integrin, which is known to function in cell motility on Fn. To test whether alpha 4 beta 1 integrins mediate cell motility on Fn and VCAM-1 in the absence of alpha 5 beta 1 integrin, we expressed human alpha 4 integrin in a Chinese hamster ovary (CHO) cell line that is deficient in alpha 5 beta 1 integrin (CHO B2). The parental alpha 5 deficient CHO B2 cells were unable to adhere, spread or migrate on Fn, nor could they assemble a fibrillar Fn matrix. Expression of alpha 4 beta 1 integrin in the CHO B2 cells enabled the cells to adhere, spread and migrate on Fn and on VCAM-1 but not to assemble a fibrillar Fn matrix. The cellular processes mediated by the interaction of alpha 4 beta 1 with Fn or VCAM-1 were inhibited by the CS1 peptide derived from the major alpha 4 beta 1 binding site on Fn. These findings demonstrate that alpha 4 beta 1 integrins not only function as cell adhesion receptors but also as cell motility receptors for Fn and VCAM-1 independent of alpha 5 beta 1. Moreover, they reveal important functional differences between Fn binding integrins. The alpha 4-positive, alpha 5-negative CHO cells described in this report will be useful tools in studying the mechanism of molecular signalling during integrin mediated cellular processes.

Amino Acid Sequence↗
PubMed1995

A novel role for alpha 3 beta 1 integrins in extracellular matrix assembly.

To study the biological role of alpha 3 beta 1 integrins in cell adhesion, migration, and in the deposition of extracellular matrix, we stably expressed the human alpha 3 integrin subunit in the alpha 4, alpha 5 integrin deficient CHO cell line B2. The expression of alpha 3 beta 1 integrins enhanced cell adhesion on entactin (also known as nidogen), but not on fibronectin. Using recombinant GST-fusion proteins that span the entire length of the entactin molecule, we located cell adhesive activity to the G2 domain of entactin. These results suggest that the alpha 3 beta 1 integrin functions as an adhesion receptor interacting with the G2 domain of entactin. On the other hand, the expression of alpha 3 beta 1 integrins did not confer the ability to migrate on entactin. Strikingly, the expression of alpha 3 beta 1 dramatically increased the deposition of entactin and fibronectin into the pericellular matrix. This was accompanied by increased binding activity of the 29 kDa amino-terminal domain of fibronectin. Thus, similar to alpha 5 beta 1 integrins, alpha 3 beta 1 integrins can play an important role in modulating the assembly of pericellular matrices. However, unlike fibronectin deposition supported by alpha 5 beta 1, alpha 3 beta 1 supported fibronectin deposition into pericellular matrix was not inhibited by antibodies binding to the RGD containing cell adhesion domain of fibronectin, demonstrating that the two processes are mechanistically distinct. The role of alpha 3 beta 1 in pericellular matrix assembly potentially implicates this receptor in the assembly and/or recognition of entactin-containing pericellular matrices, an observation consistent with its apparent role in the renal glomerulus of the mammalian kidney.

Amino Acid Sequence↗
PubMed1995

Endotoxin-induced liver injury and changes in the levels of plasma tumor necrosis factor-alpha and interleukin-6 in rabbits.

ELISA was employed to detect changes in plasma TNF-alpha and IL-6 level in rabbits having endotoxin-induced generalized Shwartzman reaction. Liver injury was assessed by the increase of serum ALT and hepatic histopathologic changes. The results showed that plasma TNF-alpha and IL-6 levels increased remarkably 12h after intravenous injection of endotoxin twice at a 24h interval, and these changes corresponded with the degree of liver injury. Coinjection of dexamethasone and endotoxin partly prevented the elevation of TNF-alpha and IL-6 levels and reduced liver injury. These results indicated that TNF-alpha and IL-6 were involved in endotoxin-induced liver injury.

Animals↗
PubMed1995

Effect of pH on myofilament Ca(2+)-sensitivity in alpha-toxin permeabilized guinea pig detrusor muscle.

PURPOSE: To ascertain if the inotropic effects of altered intracellular pH on detrusor smooth muscle could be explained by a change in Ca(2+)-sensitivity of the contractile proteins. MATERIALS AND METHODS: Guinea pig detrusor smooth muscle was permeabilized with alpha-toxin and exposed to solutions mimicking the composition of the intracellular compartment and of varying [Ca2+]. Isometric tension was measured during exposure to solutions of varying [Ca2+] at pH 6.3, 7.1 and 7.5. RESULTS: At pH 7.1 the pCa (-log10[Ca2+]) required for half-maximal activation (pCa50) was 6.00 +/- 0.07 at 22C. A Hill coefficient of unity suggested lack of cooperativity in myofilament Ca2+ activation. A decrease of pH from 7.1 to 6.3 had no significant effect on the pCa50 value or the maximum Ca2+ activated force. An increase to pH 7.5 decreased the pCa50 value by 0.65 +/- 0.20 units but left maximum force unaffected. CONCLUSIONS: The reduced Ca2+ sensitivity of detrusor myofilaments at alkaline pH could partly explain the negative inotropic effect of intracellular alkalosis in intact muscle. The positive inotropic effect of intracellular acidosis cannot, however, be explained by alteration to myofilament Ca2+ sensitivity.

Actin Cytoskeleton↗
PubMed1995

Early childhood differentials in mother-child AIDS-information interaction.

To date there have been no systematic inquiries into 1) the extent of parent-child AIDS interaction; or 2) the factors that influence whether young children and their parents talk about AIDS. In our sample, from a medium-sized Southern metropolitan area, 70 percent of mothers of children in the first, third, and fifth grades said they had talked to their child about AIDS, but only 41 percent of the mothers said their child had asked them questions about AIDS. Fourteen hypotheses about factors which might influence mother-child AIDS interaction are derived from the health/sex socialization literature. In the process of testing these bivariate hypotheses it is shown that, although sons and daughters are equally likely to ask their mother questions about AIDS, mothers are more likely to talk to their daughters than their sons about AIDS. This pattern maintains even after controls are introduced. It is suggested that discussing AIDS with young children as a health issue rather than waiting until adolescence and discussing it as a sex issue may be a more effective socialization route. Mothers may be able to do this with young daughters in the context of women's health, but because most AIDS education for adults generally is sex- rather than health-oriented they may lack the knowledge and framework to do this with their sons.

Acquired Immunodeficiency Syndrome↗
PubMed1995

[Determination of paracetmol caffeine and ephedrine hydrochloride in likeshu capsules].

Paracetamol (P), caffeine (C) and ephedrine hydrochloride (EH) are there physiological active component in Likeshu capsules. We have proposed the determination method of these three components using extractive separation-spectrophotometry. Only one amount weighed of sample is required to determine them. The recoveries of P, C and EH were 97.51 +/- 0.64% (n = 9), 98.24 +/- 1.04% (n = 8) and 98.58 +/- 1.09% (n = 9) respectively. The results of determination of samples agreed with those of the HPLC method. This method is sensitive, accurate and simple. The other ingredients of Likeshu capsules do not interfere.

Acetaminophen↗
PubMed1995

[Study on neuropeptide Y-containing innervation of cerebral small arteries and arterioles in spontaneously hypertensive rat].

To comprehend the relationship between the distribution of perivascular neuropeptides and remodified structure of arterioles in hypertension, we applied the technique of sheeting from pia matter of spontaneously hypertensive rats (SHR) to reveal branches at all levels of the vascular network so as to make an overall and stereoscopic observation of the vasculature. In addition, immunohistochemical technique and image analyser were applied to analyse qualitatively and quantitatively neuropeptide Y(NPY)-containing innervation in the vasculature. Findings revealed that the surface density of NPY-containing innervation and integral luminous density of positive reaction particle for anti-NPY in middle cerebral artery and its branches -small arteries and arterioles of SHR were significantly increased as compared with findings in Wistar Kyoto. The results reveal that an increase in density of NPY-containing innervation of cerebral small arteries and arterioles intensifies action of nutritive adjustment of vascular smooth muscle and is clearly related with increase of contractive excitability of artery, hypertrophy of artery, smooth muscle cells and the development of hypertension.

Animals↗