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C Chang

Publications and source records attributed to C Chang.

At least 37 records · Page 2Linked to original sources
PubMed1998

ETR2 is an ETR1-like gene involved in ethylene signaling in Arabidopsis.

The plant hormone ethylene regulates a variety of processes of growth and development. To identify components in the ethylene signal transduction pathway, we screened for ethylene-insensitive mutants in Arabidopsis thaliana and isolated a dominant etr2-1 mutant. The etr2-1 mutation confers ethylene insensitivity in several processes, including etiolated seedling elongation, leaf expansion, and leaf senescence. Double mutant analysis indicates that ETR2 acts upstream of CTR1, which codes for a Raf-related protein kinase. We cloned the ETR2 gene on the basis of its map position, and we found that it exhibits sequence homology to the ethylene receptor gene ETR1 and the ETR1-like ERS gene. ETR2 may thus encode a third ethylene receptor in Arabidopsis, transducing the hormonal signal through its "two-component" structure. Expression studies show that ETR2 is ubiquitously expressed and has a higher expression in some tissues, including inflorescence and floral meristems, petals, and ovules.

Amino Acid Sequence↗
PubMed1998

Association of the Arabidopsis CTR1 Raf-like kinase with the ETR1 and ERS ethylene receptors.

In Arabidopsis thaliana, signal transduction of the hormone ethylene involves at least two receptors, ETR1 and ERS, both of which are members of the two-component histidine protein kinase family that is prevalent in prokaryotes. The pathway also contains a negative regulator of ethylene responses, CTR1, which closely resembles members of the Raf protein kinase family. CTR1 is thought to act at or downstream of ETR1 and ERS based on double mutant analysis; however, the signaling mechanisms leading from ethylene perception to the regulation of CTR1 are unknown. By using the yeast two-hybrid assay, we detected a specific interaction between the CTR1 amino-terminal domain and the predicted histidine kinase domain of ETR1 and ERS. We subsequently verified these interactions by using an in vitro protein association assay(s). In addition, we determined that the amino-terminal domain of CTR1 can associate with the predicted receiver domain of ETR1 in vitro. Based on deletion analysis, the portion of CTR1 that interacts with ETR1 roughly aligns with the regulatory region of Raf kinases. These physical associations support the genetic evidence that CTR1 acts in the pathway of ETR1 and ERS and suggest that these interactions could be involved in the regulation of CTR1 activity.

Arabidopsis↗
PubMed1998

Mutations of the bacteriophage T4 type II DNA topoisomerase that alter sensitivity to antitumor agent 4'-(9-acridinylamino)methanesulfon-m-anisidide and an antibacterial quinolone.

Various antitumor and antibacterial agents target type II DNA topoisomerases, stabilizing a cleaved DNA reaction intermediate and thereby converting topoisomerase into a cellular poison. Two 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA)-resistant bacteriophage T4 topoisomerases have previously been characterized biochemically, and we have now determined the sequence of the causative mutations. In one case, a mutation (E457K) in a conserved domain of gp39 (ATPase subunit) causes resistance to antitumor agent m-AMSA but hypersensitivity to the quinolone oxolinic acid. In the second case, a combination of two amino acid substitutions (S79F and G269V) in gp52 (DNA-cleaving subunit) causes resistance to both m-AMSA and oxolinic acid. The S79F mutation is responsible for drug resistance, whereas the G269V mutation suppresses a topoisomerase deficiency caused by S79F. Surprisingly, the G269V mutation by itself causes a dramatic hypersensitivity to both inhibitors, defining a new class of topoisomerase mutants. Because S79 and the adjacent N78 are homologous to two key residues of DNA gyrase that affect quinolone sensitivity, we generated additional amino acid substitutions at these two positions. The substitutions alter sensitivity to m-AMSA and to oxolinic acid, sometimes in opposite directions. Furthermore, the quinolone sensitivities of the various mutants paralleled those of corresponding gyrase mutants. These results support models in which both quinolones and antitumor agents bind to a conserved site that overlaps the active site of the enzyme.

Amino Acid Sequence↗
PubMed1998

Cytoskeletal regulation of the signal transduction of prostaglandin EP4 receptor.

Prostaglandin (PG) EP4 receptor is coupled to Gs, stimulating adenylate cyclase. We tested whether cytoskeleton modulates the signal transduction of the EP4 receptor. A microtubule depolymerizing agent, colcemid, enhanced the PGE2-induced cAMP formation in the cloned EP4 receptor-expressing Chinese hamster ovary cells, but enhanced neither NaF plus AlCl3 nor forskolin-induced cAMP formation. Other microtubule depolymerizing agents, including colchicine, also induced the enhancement. These effects stemmed from the action of the agents on microtubules, because beta-lumicolchicine, an inactive isomer of colchicine, had no effect. In contrast, the microfilament depolymerizing agents did not affect the PGE2-induced cAMP formation but potentiated the enhancing effect of colcemid. This enhancement by colcemid was not due to the suppression of the desensitization of the EP4 receptor. The enhancing effect of colcemid was also observed in another Gs-coupled PGE receptor subtype, EP2 receptor. These results demonstrate that the state of microtubule assembly modulates the signal transduction of the EP4 receptor in concert with microfilament.

Adenylyl Cyclases↗
PubMed1998

Expression of human prostatic acid phosphatase correlates with androgen-stimulated cell proliferation in prostate cancer cell lines.

Androgen plays a critical role in regulating the growth and differentiation of normal prostate epithelia, as well as the initial growth of prostate cancer cells. Nevertheless, prostate carcinomas eventually become androgen-unresponsive, and the cancer is refractory to hormonal therapy. To gain insight into the mechanism involved in this hormone-refractory phenomenon, we have examined the potential role of the androgen receptor (AR) in that process. We have investigated the expression of AR and two prostate-specific androgen-responsive antigens, prostatic acid phosphatase (PAcP) and prostate-specific antigen (PSA), for the functional activity of AR in LNCaP and PC-3 human prostate carcinoma cells. Our results are as follows. (i) Clone 33 LNCaP cells express AR, PAcP, and PSA, and cell growth is stimulated by 5alpha-dihydrotestosterone (DHT). Stimulation of cell growth correlates with decreased cellular PAcP activity. (ii) In clone 81 LNCaP cells, the expression of PAcP decreases with a concurrent decrease in the degree of androgen stimulation of cell growth, whereas the expression of PSA mRNA level is up-regulated by DHT, as in clone 33 cells. Conversely, in PAcP cDNA-transfected clone 81 cells, an additional expression of cellular PAcP correlates with an increased stimulation by androgen, higher than the corresponding control cells. (iii) PC-3 cells express a low level of functional AR with no detectable PAcP or PSA, and the growth of PC-3 cells is not affected by DHT treatment. Nevertheless, in two PAcP cDNA-transfected PC-3 sublines, the expression of exogenous cellular PAcP correlates with androgen stimulation. This androgen stimulation of cell growth concurs with an increased tyrosine phosphorylation of a phosphoprotein of 185 kDa. In summary, the data indicate that the expression of AR alone is not sufficient for androgen stimulation of cell growth. Furthermore, in AR-expressing prostate cancer cells, the expression of cellular PAcP correlates with androgen stimulation of cell proliferation.

Acid Phosphatase↗
PubMed1998

Neural crest induction by Xwnt7B in Xenopus.

Neural patterning occurs soon after neural induction during early development. In Xenopus, several caudalizing factors transform anterior neural to posterior neural tissue at the open neural plate stages, while other factors are responsible for setting up mediolateral polarity which becomes the dorsoventral (D-V) axis after neural tube closure. Many Wnt ligands are expressed in the neural tube in distinct anteroposterior (A-P) and D-V domains, implying a function in neural patterning. Here we report the cloning of a full-length Xenopus Wnt7B gene. Xwnt7B induces neural crest markers Xslug and Xtwist in ectodermal explants coinjected with neural inducer noggin and in ectodermal cells neuralized by dissociation. In vivo, Xwnt7B expands the Xtwist expression domain when injected in the animal pole. Our results suggest that Wnt members are involved in dorsoventral patterning of the neural tube.

Amino Acid Sequence↗
PubMed1998

Human aldehyde dehydrogenase gene family.

Twelve aldehyde dehydrogenase (ALDH) genes have been identified in humans. These genes, located on different chromosomes, encode a group of enzymes which oxidizes varieties of aliphatic and aromatic aldehydes. Metabolic disorders and clinical problems associated with mutations of ALDH1, ALDH2, ALDH4, ALDH10 and succinic semialdehyde (SSDH) genes have been emerged. Comparison of the human ALDHs indicates a wide range of divergency (> 80 - < 15% identity at the protein sequence level) among them. However, several protein regions, some of which are implicated in functional activities, are conserved in the family members. The phylogenic tree constructed of 56 ALDH sequences of humans, animals, fungi, protozoa and eubacteria, suggests that the present-day human ALDH genes were derived from four ancestral genes that existed prior to the divergence of Eubacteria and Eukaryotes. The neighbor-joining tree derived from 12 human ALDHs and antiquitin indicates that diversification within the ALDH1/2/5/6 gene cluster occurred during the Neoproterozoic period (about 800 million years ago). Duplication in the ALDH 3/10/7/8 gene cluster occurred in Phanerozoic period (about 300 million years ago). Separations of ALDH3/ALDH10 and that of ALDH7/ALDH8 had occurred during the period of appearance and radiation of mammalian species.

Aldehyde Dehydrogenase↗
PubMed1998

Constitutive and inducible hsp70s are involved in oxidative resistance evoked by heat shock or ethanol.

Improved cardiac post-ischemic recovery after whole-body hyperthermia is correlated with an increased expression of the heat shock proteins (hsps). The inducible hsp70 (hsp70i) has a known cardioprotective effect against ischemia/reperfusion injury. Here, we studied whether other hsps are also involved in cardioprotection. Using rat heart-derived H9c2 myocytes, we observed that preheating at 43 degreesC for 20 min conferred resistance to hydrogen peroxide (H2O2). The resistance to mild H2O2 toxicity (3-5 micro mol/10(7) cells) appeared early and persisted, whereas the resistance to moderate H2O2 toxicity (6-9 micro mol/10(7) cells) was detectable only at 20-44 h post heat shock. No resistance was observed at higher doses of hydrogen peroxide (10-12 micro mol/10(7) cells), indicating that severe toxicity exceeds the capacity of the induced protective mechanism. Coincidentally, this thermal regimen elicited a rapid and prolonged increase in the cellular level of hsp70i, and a delayed and transient induction of the constitutive hsp70 (hsp70c). Nuclear translocations of hsp70i and hsp70c also occurred upon heat stimulation. A homogeneous distribution of the accumulated hsp70i and hsp70c throughout the nuclei and cytoplasm paralleled the development of heat-induced resistance to moderate H2O2 challenge. Application of another hsp inducer, ethyl alcohol, evoked a similar pattern of H2O2 resistance, and hsp induction and distribution. Our results suggest that induction and subcellular distribution of hsp70s contribute importantly to cellular antioxidant defenses, and that a co-operation between hsp70i and hsp70c may improve cardiac preservation during oxidative insult.

Animals↗
PubMed1998

Stable overexpression of the constitutive form of heat shock protein 70 confers oxidative protection.

We have previously reported that thermal preconditioning confers an oxidative resistance in rat heart-derived H9c2 myocytes. The development of this resistance is associated with a co-expression of both inducible (hsp70i) and constitutive (hsp70c) forms of the 70-kD heat shock proteins, suggesting an antioxidant role for these proteins. Overexpression of hsp70i has been shown to render cells more tolerant to oxidative challenge. The present study sought to determine whether increases in hsp70c, the constitutive member of this protein family, are also positively correlated to oxidative protection. A rat cDNA encoding hsp70c was inserted into a mammalian expression vector, allowing transcription of the inserted gene to be regulated by a powerful cytomegaloviral promoter. After introduction of this construct into H9c2 myocytes, stable clones were obtained. Western and Northern blot analysis of the derived clones showed a two-fold increase in hsp70c mRNA and protein concentrations. These clones were more resistant to thermal killing when compared to control cells transfected with the vector alone, implicating a functional role for the overexpressed hsp70c protein. hsp70c-enriched cells also exhibited a marked resistance to oxidative challenges, including exposure to hydrogen peroxide (H2O2), hydroxyl radical, menadione, and hypoxia/reoxygenation. These findings indicate that hsp70c overexpression provides a protective effect against endogenous or exogenously generated reactive oxygen species (ROS), suggesting that hsp70c actively participates in the heat shock-induced oxidative protection.

Animals↗
PubMed1998

Direct quantitation of HIV by flow cytometry using branched DNA signal amplification.

Adaptation of the branched DNA signal amplification technology to flow cytometry has resulted in a quantitative nuclei-acid assay with significant advantages over the microwell-based format. In this assay, microbeads, rather than microwell plates, are derivatized with nucleic-acid capture probes and the derivatized beads are used to capture single nucleic-acid targets, which then capture fluorescent reporter probes via branched DNA. The assay detects DNA or RNA targets, has a current lower sensitivity limit of 500 human immunodeficiency virus (HIV) RNA molecules and responds linearly to target level from 500 to at least 50,000 molecules. Since microbeads can easily interrogate large volumes, viral lysis and genomic RNA capture can proceed in one step from comparatively large volumes, and sample preparation is greatly simplified compared to the microwell-format bDNA assay.

Cell Line↗
PubMed1998

The molecular basis of C4 photosynthesis in sorghum: isolation, characterization and RFLP mapping of mesophyll- and bundle-sheath-specific cDNAs obtained by differential screening.

C4 photosynthesis depends upon the strict compartmentalization of the CO2-assimilatory enzymes of the C4 and Calvin cycle in two different cell types, mesophyll and bundle-sheath cells. A differential accumulation is also observed for enzymes of other metabolic pathways, and mesophyll and bundle-sheath chloroplasts of NADP-malic enzyme type C4 plants differ even in their photosynthetic electron transport chains. A large number of studies indicate that this division of labour between mesophyll and bundle-sheath cells is the result of differential gene expression. To investigate the extent of this differential gene expression and thus gain insight into the genetic basis of C4 photosynthesis, genes that are differentially expressed in the mesophyll and bundle-sheath cells were catalogued in the NADP-malic enzyme type C4 grass Sorghum bicolor. A total of 58 cDNAs were isolated by differential screening. Using a tenfold difference in transcript abundance between mesophyll and bundle-sheath cells as a criterion, 25 cDNAs were confirmed to encode mesophyll-specific gene sequences and 8 were found to encode bundle-sheath-specific sequences. Eight mesophyll-specific cDNAs showed no significant similarities within GenBank and may therefore represent candidates for the elucidation of hitherto unknown functions in the differentiation of mesophyll and bundle-sheath cells. The chromosomal location of 50 isolated cDNAs was determined by RFLP mapping using an interspecific sorghum cross.

Base Sequence↗
PubMed1998

Thyroid hormone direct repeat 4 response element is a positive regulatory element for the human TR2 orphan receptor, a member of steroid receptor superfamily.

We demonstrate that TR2 orphan receptor (TR2) may induce transactivation activities via an AGGTCA-like-direct-repeat-4 consensus thyroid hormone response element (DR4-TRE) system. TR2 showed a slightly greater binding affinity than thyroid hormone receptor alpha1 (TR alpha1)/retinoid X receptor alpha (RXR alpha) heterodimer with Kds 0.5 nM and 2.3 nM, respectively. These receptors, TR2 and TR alpha1/RXR alpha heterodimer, competed with each other on binding to limited amounts of DR4-TRE. TR2 canceled the suppression effect of unliganded-TR alpha1 on CAT reporter activity in a dose-dependent fashion. Estrogen receptor (ER) and 2P2 (a mutated TR2 with P box sequence of androgen receptor) failed not only to bind to DR4-TRE but also to recover this inhibitory effect of unliganded TRalpha1. However, when T3 was supplemented, estradiol-ER competed for a full CAT activity while TR2 showed an additive effect on the transcriptional activation. These results indicate that DNA binding is essential for TR2 to take action and fully functional liganded TR alpha1 may rely on common factors shared with ER but not TR2.

Carrier Proteins↗
PubMed1998

Anti-tumor effects of d-dicentrine from the root of Lindera megaphylla.

d-Dicentrine, a naturally occurring aporphine type isoquinoline alkaloid, isolated from the root of Lindera megaphylla Hemsl. (Lauraceae), was evaluated for its potential anti-cancer activity. We found d-dicentrine significantly inhibited the growth of human hepatoma cell line HuH-7 by delaying its doubling time in tissue culture. An in vitro colony forming assay showed that d-dicentrine decreased the colony formation efficiency in both hepatoma cell lines, HuH-7 and MS-G2, used in our study. Biosyntheses of the macromolecules DNA and RNA were also strongly inhibited. An MTT assay in 21 tumor cell lines also revealed that d-dicentrine was most cytotoxic to esophageal carcinoma HCE-6, lymphoma cell lines Molt-4 and CESS, leukemia cell lines HL60 and K562, and hepatoma cell line MS-G2. An in vitro tumor growing assay in the Severe Combined immunodeficiency (SCID) mice showed that intraperitoneal injection of d-dicentrine at the dose of 100 micrograms twice a week for 4 weeks significantly inhibited the tumor incidence of leukemia cell line K562 in SCID mice. All these data indicated that d-dicentrine has potential anti-tumor applications.

Animals↗
PubMed1998

Characterization of fibrous retrocorneal membrane by confocal microscopy.

PURPOSE: To study the appearance of a fibrous retrocorneal membrane as seen by confocal microscopy. METHODS: A 67-year-old white woman with a history of multiple ocular surgeries, including repeated penetrating keratoplasties for aphakic bullous keratopathy, developed a retrocorneal membrane in the right eye. The membrane was first noticed 3 years after the last corneal transplant and remained stable subsequently. The patient was examined by in vivo white light tandem-scanning confocal microscopy. RESULTS: At the level of the retrocorneal membrane, confocal microscopy disclosed the presence of a hyperreflective fibrous-appearing layer. Normal endothelial cells could not be found. Anterior to the hyperreflective layer, activated keratocytes were identified. CONCLUSION: Confocal microscopy may allow noninvasive diagnosis of fibrous retrocorneal membrane. Additionally, our data suggest that the posterior keratocytes might play a role in the production and deposition of fibrous tissue.

Aged↗
PubMed1998

Differential expression of Nur77 family members in human T-lymphotropic virus type 1-infected cells: transactivation of the TR3/nur77 gene by Tax protein.

We analyzed the differential expression and regulation of three members of the Nur77 transcription factor family by the human T-lymphotropic virus type 1 (HTLV-1) Tax protein. We have demonstrated that in both HTLV-1-infected cells and Tax-expressing JPX-9 cells, TR3/nur77 is highly expressed, whereas neither NOR-1 nor NOT expression is detectable. Transient transfection analysis further confirmed the Tax transactivation of the TR3/nur77 promoter but not the NOR-1 promoter in different cell types. Furthermore, expression of a luciferase reporter gene driven by the NGFI-B (rat homolog of TR3/Nur77) response element (NBRE) provided evidence that Tax-mediated transactivation resulted in the induction of a functional protein. Cotransfection assays with the TR3/nur77 promoter sequence or the NBRE binding motif together with a series of Tax mutants have shown that Tax-induced TR3/nur77 expression is mediated by CREB/ATF-related transcription factors.

Activating Transcription Factor 1↗
PubMed1998

Mononeuropathy multiplex in hemophilia: an electrophysiologic assessment.

Single nerve lesion or mononeuropathy caused by extrinsic compression of hematoma or hemarthrosis is consistently reported for hemophilic patients. To study the manifestation of peripheral nerve lesions, an extensive nerve conduction study of peripheral nerves and specific nerves in correlation with clinical involvement was carried out on 126 hemophilic patients. We found 24 patients with 34 peripheral nerve lesions. The frequency of nerve lesion positively correlated with the severity of hemophilia (p < 0.01) and increasing age (p < 0.05). Nerve lesions occurred more commonly in lower extremities than in upper extremities. The femoral nerve was most commonly involved. Seven patients were identified to have mononeuropathy multiplex with 2 or more nerve lesions involving more than one extremity. All of these patients had chronic condition. Our findings suggest that mononeuropathy multiplex may occur in hemophilic patients.

Adolescent↗
PubMed1998

Antisense TR3 orphan receptor can increase prostate cancer cell viability with etoposide treatment.

In androgen-responsive LNCaP human prostatic cancer cells, human TR3 orphan receptor, a member of the steroid receptor superfamily, can be rapidly induced by androgen. In contrast, ablation of androgen by castration can induce the expression of the TR3 orphan receptor gene in rat ventral prostate that has undergone apoptosis. This phenomenon prompted us to further analyze the potential role of human TR3 orphan receptor in prostate cancer cells in which apoptosis had been induced. Northern blot analysis shows that human TR3 orphan receptor expression can be induced rapidly after treatment of LNCaP and PC-3 prostate cancer cells with calcium ionophore or etoposide. Our data further demonstrate that a much higher concentration of etoposide was needed to kill the same number of cells in LNCaP and PC-3 cells transfected stably with antisense TR3 orphan receptor compared with that in control vector transfectants. Together, our data suggest that the human TR3 orphan receptor may play an important role in modulating drug-induced prostate apoptosis.

Androgens↗