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W L Yang

Publications and source records attributed to W L Yang.

54 records · Page 3Linked to original sources
PubMed1994

Purification and characterization of CTP synthetase, the product of the URA7 gene in Saccharomyces cerevisiae.

In the yeast Saccharomyces cerevisiae, CTP synthetase [EC 6.3.4.2; UTP:ammonia ligase (ADP-forming)] is the product of the URA7 gene. CTP synthetase was purified 503-fold to apparent homogeneity from cells bearing the URA7 gene on a multicopy plasmid that directed a 10-fold overproduction of the enzyme. The purification procedure included ammonium sulfate fractionation of the cytosolic fraction followed by chromatography with Sephacryl 300 HR, Q-Sepharose, Affi-Gel Blue, and Superose 6. The N-terminal amino acid sequence of purified CTP synthetase was identified and aligned perfectly with the deduced sequence of the URA7 gene. The minimum subunit molecular mass (68 kDa) of purified CTP synthetase was in good agreement with the size (64.7 kDa) of the URA7 gene product. Antibodies were raised against a maltose-binding protein-CTP synthetase fusion protein which immunoprecipitated CTP synthetase from wild-type cells. Immunoblot analysis was used to identify CTP synthetase in wild-type cells and cells bearing the URA7 gene on a multicopy plasmid. The results of gel filtration chromatography indicated that the size of native CTP synthetase was consistent with a dimeric structure for the enzyme. CTP synthetase oligomerized to a tetramer in the presence of its substrates UTP and ATP. Maximum CTP synthetase activity was dependent on magnesium ions (4 mM) and 2-mercaptoethanol at the pH optimum of 8.0. CTP synthetase exhibited positive cooperative kinetics with respect to UTP and ATP and negative cooperative kinetics with respect to glutamine and GTP. CTP synthetase was potently inhibited by the product CTP which also increased the positive cooperativity of the enzyme toward UTP.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗
PubMed1994

Use of synthetic lethal mutants to clone and characterize a novel CTP synthetase gene in Saccharomyces cerevisiae.

In the pyrimidine biosynthetic pathway, CTP synthetase catalyses the conversion of uridine 5'-triphosphate (UTP) to cytidine 5'-triphosphate (CTP). In the yeast Saccharomyces cerevisiae, the URA7 gene encoding this enzyme was previously shown to be nonessential for cell viability. The present paper describes the selection of synthetic lethal mutants in the CTP biosynthetic pathway that led us to clone a second gene, named URA8, which also encodes a CTP synthetase. Comparison of the predicted amino acid sequences of the products of URA7 and URA8 shows 78% identity. Deletion of the URA8 gene is viable in a haploid strain but simultaneous presence of null alleles both URA7 and URA8 is lethal. Based on the codon bias values for the two genes and the intracellular concentrations of CTP in strains deleted for one of the two genes, relative to the wild-type level, URA7 appears to be the major gene for CTP biosynthesis. Nevertheless, URA8 alone also allows yeast growth, at least under standard laboratory conditions.

Amino Acid Sequence↗
PubMed1992

Structural basis of the in vivo induction of micronuclei.

The structural basis of the in vivo induction of micronuclei was examined with CASE, a structure-activity relational method. The CASE program identified a number of structures associated with this activity. When used to predict the activity of chemicals not included in the learning set, these structural determinants gave a concordance in excess of 83%. The existence of a structural basis for the induction of micronuclei will permit an investigation of the mechanistic basis of this phenomenon.

Databases, Factual↗
PubMed1991

[Surgical treatment of superior mesenteric artery syndrome].

Fourty-two patients with superior mesenteric artery syndrome were treated surgically. Among the patients, 24 were male and 18 female (mean age, 38 years). The courses of the disease varied from 1 to 10 years. Operations included amputation of the ligament of Treitz (14 patients), gastrojejunostomy (2), subtotal gastrectomy and gastro jejunostomy (1), duodenojejunostomy (6), anterior repositioning of the duodenum (7), and circular drainage operation of the duodenum (12). Two patients underwent anterior repositioning of the duodenum and gastrojejunostomy. In patients without relief of symptoms such as vomiting after operation, circular drainage of the duodenum was performed. All the 42 patients were successfully treated. The authors discussed 6 kinds of operations, their indications, and the advantages and disadvantages. Emphasis was given to the anterior repositioning of the duodenum and its circular drainage.

Adolescent↗
PubMed1989

[Protective effect of extracts from Aloe vera L. var. chinensis (Haw.) Berg. on experimental hepatic lesions and a primary clinical study on the injection of in patients with hepatitis].

The injection(10-15 ml/kg/d, ip x 4), total glycoside (125-225 mg/kg/d, ip x (3-4); 600 mg/kg/d, ig x 3) and crystal III (120 mg/kg/d, ip x 4) of Aloe vera var. chinensis were found to be effective in lowering the elevated sGPT induced by CCl4, thioacetamide and D-aminogalactose in mice or rats. It was also observed that these agents could protect hepatic cells from the CCl4-induced injury. When dogs were given in with Aloe injection of 0.1 ml/kg/d x 180, no toxicity was noted. The total effective sGPT-lowering rate of Aloe injection on 38 patients of chronic hepatitis with positive HBsAg was 86.8%.

Alanine Transaminase↗
PubMed1989

[Histologic distribution of anti-glioma monoclonal antibody SZ-39 in human xenograft brain tumors].

131I-labeled anti-glioma monoclonal antibody SZ-39 was injected intraperitoneally into nude mice bearing xenograft human glioma, ependymoma and intracranial metastatic adenocarcinoma. Subsequent gamma-imaging and assessment of radioactivity showed selective accumulation of antibody in the glioma. Quantitative autoradiography supported the results of radiolocalization observed in vivo in different brain tumors and normal tissues. It was also shown that antibody SZ-39 was specially localized to viable glioma cells and closely associated with their cell membrane. Yet, it was not bound to fibrous or necrotic areas. 131I-normal mouse IgG was distributed in a nonspecific pattern in the glioma. These studies demonstrated the specificity of SZ-39 in vivo which may be useful in clinical diagnostic guiding and treatment.

Adenocarcinoma↗
PubMed1987

Presence of hepatitis B virus DNA in serum of surface-antigen-seronegative immunocompromised patients.

We compared the prevalence of serum hepatitis B virus (HBV) DNA in cancer patients who were immunocompromised with that in healthy subjects. Testing positive for serum HBV DNA were 27 of the 232 (11.6%) surface-antigen-negative cancer patients and 7 of the 382 (1.8%) surface-antigen-negative healthy subjects. These data suggest that the negative serology, at least in immunocompromised individuals, is probably not sufficient to exclude HBV infection.

DNA, Viral↗
PubMed1987

Hepatitis type B virus DNA in patients receiving hemodialysis: correlation with other HBV serological markers.

Possible presence of hepatitis type B virus (HBV) was assessed in 239 end-stage renal failure patients who were receiving long-term maintenance hemodialysis (average 30.8 months; duration: 1-94 months), and who had not shown any other symptom of HBV infection. Their HBV serological markers, including HBV DNA, were evaluated together with those of normal control individuals. HBV surface antigen (HBsAg) was detected in 42 of the 239 dialysis patients, 15 of whom also positive for HBV DNA (mean +/- SD = 56.2 +/- 23.7 pg/100 microliters of serum). HBV DNA was also found in 22 of the 197 (11.2%) dialysis patients who were negative for HBsAg, with an average of 36.2 +/- 19.0 pg/100 microliters of serum. This rate of detecting HBV DNA in HBV seronegative dialysis individuals was significantly higher than the rate of 1.83% found among healthy HBsAg(-) individuals. Among these 22 dialysis patients who were HBsAg(-) but HBV DNA(+), 15 were found to possess antibodies against HBsAg (anti-HBs) and/or antibody against HBV e antigen (anti-HBe). These data suggested that the absence of serum HBV antigen or the presence of antibodies against HBV markers might not be sufficient to identify possible HBV infection in immunocompromised hosts such as hemodialysis patients.

Adult↗
PubMedDate not supplied

Cisplatin sensitivity in cAMP-dependent protein kinase mutants of Saccharomyces cerevisiae.

The emergence of cisplatin resistance poses a significant problem to the treatment of a variety of human malignancies. Therefore, understanding the molecular basis of cisplatin resistance could improve the clinical effectiveness of this anticancer agent. Recently, our laboratory has demonstrated that cAMP-dependent protein kinase (PKA) mutants of the Chinese hamster ovary (CHO) and the mouse adrenocortical carcinoma Y1 cells exhibited increased resistance to cisplatin as well as other DNA-damaging drugs. Further studies showed that either the functional inactivation of PKA or the mutation in the regulatory subunit gene may cause increased recognition of cisplatin-damaged DNA and enhanced DNA repair capacity. In this study, we evaluated the role of PKA in modulating cellular sensitivity to cisplatin in a series of PKA mutants of Saccharomyces cerevisiae. Mutants with decreased kinase activity resulting from a srv2 mutation showed no alterations in cisplatin sensitivity. Complementation of TPK1 in a yeast strain containing mutant tpk1 and also tpk2 and tpk3 deletions did not significantly alter its sensitivity to this DNA-damaging agent. Yeast transformants containing increased kinase activity resulting from overexpression of RAS2Val19 or TPK1 and yeast strains having increased kinase activities due to mutations in the BCY1 gene also did not show alterations in their sensitivity to cisplatin. Therefore, results from these studies unambiguously demonstrate that changes in PKA activity have no effect on cisplatin sensitivity in Saccharomyces cerevisiae.

Antineoplastic Agents↗