Biomedical subjects

G Feng

Publications and source records attributed to G Feng.

At least 19 recordsLinked to original sources

PubMed1996

Cloning and functional characterization of a novel dopamine receptor from Drosophila melanogaster.

A cDNA clone is described that encodes a novel G-protein-coupled dopamine receptor (DopR99B) expressed in Drosophila heads. The DopR99B receptor maps to 99B3-5, close to the position of the octopamine/tyramine receptor gene at 99A10-B1, suggesting that the two may be related through a gene duplication. Agonist stimulation of DopR99B receptors expressed in Xenopus oocytes increased intracellular Ca2+ levels monitored as changes in an endogenous inward Ca2+-dependent chloride current. In addition to initiating this intracellular Ca2+ signal, stimulation of DopR99B increased cAMP levels. The rank order of potency of agonists in stimulating the chloride current is: dopamine > norepinephrine > epinephrine > tyramine. Octopamine and 5-hydroxytryptamine are not active (< 100 microM). This pharmacological profile plus the second-messenger coupling pattern suggest that the DopR99B receptor is a D1-like dopamine receptor. However, the hydrophobic core region of the DopR99B receptor shows almost equal amino acid sequence identity (40-48%) with vertebrate serotonergic, alpha 1- and beta-adrenergic, and D1-like and D2-like dopaminergic receptors. Thus, this Drosophila receptor defines a novel structural class of dopamine receptors. Because DopR99B is the second dopamine receptor cloned from Drosophila, this work establishes dopamine receptor diversity in a system amenable to genetic dissection.

Amino Acid Sequence↗

PubMed1996

Amino acid substitutions in the two largest subunits of Escherichia coli RNA polymerase that suppress a defective Rho termination factor affect different parts of the transcription complex.

Among the earliest rpoBC mutations identified are three suppressors of the conditional lethal rho allele, rho201. These three mutations are of particular interest because, unlike rpoB8, they do not increase termination at all rho-dependent and rho-independent terminators. rpoB211 and rpoB212 both change Asn-1072 to His in conserved region H of rpoB (betaN1072H), whereas rpoC214 changes Arg-352 to Cys in conserved region C of rpoC (beta'R352C). Both substitutions significantly reduce the overall rate of transcript elongation in vitro relative to wild-type RNA polymerase; however, they probably slow elongation for different reasons. The nucleotide triphosphate concentrations required at the T7 A1 promoter for both abortive trinucleotide synthesis and for promoter escape are much greater for betaN1072H. In contrast, beta'R352C and two adjacent substitutions (beta'G351S and beta'S350F), but not betaN1072H, formed open complexes of greatly reduced stability. The sequence in this region of beta' modestly resembles a region of Escherichia coli DNA polymerase I that contacts the phosphate backbone of DNA in co-crystals. Core determinants affecting open complex formation do not reside exclusively in beta', however, since the Rifr mutation rpoB2 in beta also dramatically destabilized open complexes. We suggest that the principal defects of the two Rho-suppressing substitutions may differ, perhaps reflecting a greater role of beta region H in nucleoside triphosphate-binding and nucleotide addition and of beta' region C in contacts to the DNA strands that could be important for translocation. Although both probably suppress rho201 by slowing RNA chain elongation, these differences may lead to terminator specificity that depends on the rate-limiting step at different sites.

Amino Acid Sequence↗

PubMed1996

Some acoustic features of nasal and nasalized vowels: a target for vowel nasalization.

In order to characterize acoustic properties of nasal and nasalized vowels, these sounds will be considered as a dynamic trend from an oral configuration toward an [n]-like configuration. The latter can be viewed as a target for vowel nasalization. This target corresponds to the pharyngonasal tract and it can be modeled, with some simplifications, by a single tract without any parallel paths. Thus the first two resonance frequencies (at about 300 and 1000 Hz) characterize this target well. A series of measurements has been carried out in order to describe the acoustic characteristics of the target. Measured transfer functions confirm the resonator nature of the low-frequency peak. The introduction of such a target allows the conception of the nasal vowels as a trend beginning with a simple configuration, which is terminated in the same manner, so allowing the complex nasal phenomena to be bounded. A complete study of pole-zero evolutions for the nasalization of the 11 French vowels is presented. It allows the proposition of a common strategy for the nasalization of all vowels, so a true nasal vowel can be placed in this nasalization frame. The measured transfer functions for several French nasal vowels are also given.

France↗

PubMed1996

Depletion of the cellular amounts of the MutS and MutH methyl-directed mismatch repair proteins in stationary-phase Escherichia coli K-12 cells.

The MutL, MutS, and MutH proteins mediate methyl-directed mismatch (MDM) repair and help to maintain chromosome stability in Escherichia coli. We determined the amounts of the MDM repair proteins in exponentially growing, stationary-phase, and nutrient-starved bacteria by quantitative Western immunoblotting. Extracts of null mutants containing various amounts of purified MDM repair proteins were used as quantitation standards. In bacteria growing exponentially in enriched minimal salts-glucose medium, about 113 MutL dimers, 186 MutS dimers, and 135 MutH monomers were present per cell. Calculations with the in vitro dissociation constants of MutS binding to different mismatches suggested that MutS is not present in excess, and may be nearly limiting in some cases, for MDM repair in exponentially growing cells. Remarkably, when bacteria entered late stationary phase or were deprived of a utilizable carbon source for several days, the cellular amount of MutS dropped at least 10-fold and became barely detectable by the methods used. In contrast, the amount of MutH dropped only about threefold and the amount of MutL remained essentially constant in late-stationary-phase and carbon-starved cells compared with those in exponentially growing bacteria. RNase T2 protection assays showed that the amounts of mutS, mutH, and mutL, but not miaA, transcripts decreased to undetectable levels in late-stationary-phase cells. These results suggested that depletion of MutS in nutritionally stressed cells was possibly caused by the relative instability of MutS compared with MutL and MutH. Our findings suggest that the MDM repair capacity is repressed in nutritionally stressed bacteria and correlate with conclusions from recent studies of adaptive mutagenesis. On the other hand, we did not detect induction of MutS or MutL in cells containing stable mismatches in multicopy single-stranded DNA encoded by bacterial retrons.

Adenosine Triphosphatases↗

PubMed1995

Cloning and functional analysis of TipE, a novel membrane protein that enhances Drosophila para sodium channel function.

Voltage-dependent sodium channels are involved in the initiation and propagation of action potentials in many excitable cells. Here we report that tipE, a gene defined by a temperature-sensitive paralytic mutation in Drosophila, encodes a novel integral membrane protein that dramatically stimulates functional expression in Xenopus oocytes of the Drosophila sodium channel alpha subunit encoded by the paralytic (para) locus. Using a heat shock promoter to control tipE+ gene expression in transgenic flies, we demonstrate that tipE+ gene expression is required during pupal development to rescue adult paralysis. In addition, we demonstrate a role for the tipE gene product in adults.

Amino Acid Sequence↗

PubMed1995

Discontinuous movements of DNA and RNA in RNA polymerase accompany formation of a paused transcription complex.

A central enigma of transcriptional regulation is how the normally efficient transcription elongation complex stops at pause and termination signals. One possibility, raised by the discovery that RNA polymerase sometimes contracts its DNA footprint, is that discontinuous movements contribute to recognizing these signals. We report that E. coli RNA polymerase responds to sequences immediately downstream and upstream from the his leader pause site by changing neither its downstream DNA contact nor its upstream RNA contact for 8 bp preceding the pause. This compressed complex isomerizes to a paused conformation by an approximately 10 bp jump of its downstream DNA contact and simultaneous extrusion of an RNA hairpin that stabilizes the paused conformation. We suggest pausing and termination could be alternative outcomes of a similar isomerization that depend on the strength of contacts to 3'-proximal RNA remaining after the jump.

Base Sequence↗

PubMed1995

Cytogenetic and molecular localization of tipE: a gene affecting sodium channels in Drosophila melanogaster.

Voltage-sensitive sodium channels play a key role in nerve cells where they are responsible for the increase in sodium permeability during the rising phase of action potentials. In Drosophila melanogaster a subset of temperature-sensitive paralytic mutations affect sodium channel function. One such mutation is temperature-induced paralysis locus E (tipE), which has been shown by electrophysiology and ligand binding studies to reduce sodium channel numbers. Three new gamma-ray-induced tipE alleles associated with either visible deletions in 64AB or a translocation breakpoint within 64B2 provide landmarks for positional cloning of tipE. Beginning with the flanking cloned gene Ras2, a 140-kb walk across the translocation breakpoint was completed. Germline transformation using a 42-kb cosmid clone and successively smaller subclones localized the tipE gene within a 7.4-kb genomic DNA segment. Although this chromosome region is rich in transcripts, only three overlapping mRNAs (5.4, 4.4, and 1.7 kb) lie completely within the smallest rescuing construct. The small sizes of the rescuing construct and transcripts suggest that tipE does not encode a standard sodium channel alpha-subunit with four homologous repeats. Sequencing these transcripts will elucidate the role of the tipE gene product in sodium channel functional regulation.

Alleles↗

PubMed1995

Cloning and characterization of a calcium channel alpha 1 subunit from Drosophila melanogaster with similarity to the rat brain type D isoform.

We report the complete sequence of a calcium channel alpha 1 subunit cDNA cloned from a Drosophila head cDNA library. This cDNA encodes a deduced protein containing 2516 amino acids with a predicted molecular weight of 276,493. The deduced protein shares many features with vertebrate homologs, including four repeat structures, each containing six transmembrane domains, a conserved ion selectivity filter region between transmembrane domains 5 and 6, and an EF hand in the carboxy tail. The Drosophila subunit has unusually long initial amino and terminal carboxy tails. The region corresponding to the last transmembrane domain (IVS6) and the adjacent cytoplasmic domain has been postulated to form a phenylalkylamine-binding site in vertebrate calcium channels. This region is conserved in the Drosophila sequence, while domains thought to be involved in dihydropyridine binding show numerous changes. The Drosophila subunit exhibits 78.3% sequence similarity to the rat brain type D calcium channel alpha 1 subunit, and so has been designated as a Drosophila melanogaster calcium channel alpha 1 type D subunit (Dmca1D). In situ hybridization shows that Dmca1D is highly expressed in the embryonic nervous system. Northern analysis shows that Dmca1D cDNA hybridizes to three size classes of mRNA (9.5, 10.2, and 12.5 kb) in heads, but only two classes (9.5 and 12.5 kb) in bodies and legs. PCR analysis suggests that the Dmca1D message undergoes alternative splicing with more heterogeneity appearing in head and embryonic extracts than in bodies and legs.

Amines↗

PubMed1995

Radiologic manifestations of osteopetrosis.

The clinical radiologic manifestations of 8 patients with osteopetrosis (4 males and 4 females) are analyzed. Six of the cases are benign. The pathologic basis for the radiologic manifestations is discussed, and a classification is suggested. "Broom-like" metaphyses, metaphyses containing three or four dense layers of bone, and ossification of soft tissues around joints were found and differential diagnosis was made.

Adolescent↗

PubMed1994

Termination-altering amino acid substitutions in the beta' subunit of Escherichia coli RNA polymerase identify regions involved in RNA chain elongation.

To identify regions of the largest subunit of RNA polymerase that are potentially involved in transcript elongation and termination, we have characterized amino acid substitutions in the beta' subunit of Escherichia coli RNA polymerase that alter expression of reporter genes preceded by terminators in vivo. Termination-altering substitutions occurred in discrete segments of beta', designated 2, 3a, 3b, 4a, 4b, 4c, and 5, many of which are highly conserved in eukaryotic homologs of beta'. Region 2 substitutions (residues 311-386) are tightly clustered around a short sequence that is similar to a portion of the DNA-binding cleft in E. coli DNA polymerase I. Region 3b (residues 718-798) corresponds to the segment of the largest subunit of RNA polymerase II in which amanitin-resistance substitutions occur. Region 4a substitutions (residues 933-936) occur in a segment thought to contact the transcript 3' end. Region 5 substitutions (residues 1308-1356) are tightly clustered in conserved region H near the carboxyl terminus of beta'. A representative set of mutant RNA polymerases were purified and revealed unexpected variation in percent termination at six different rho-independent terminators. Based on the location and properties of these substitutions, we suggest a hypothesis for the relationship of subunits in the transcription complex.

Amino Acid Sequence↗

PubMed1994

Crystallization of GreA, a transcript cleavage factor from Escherichia coli.

GreA is a 17.6 kDa protein from Escherichia coli that induces cleavage of the nascent transcript in the elongating complex of RNA polymerase, followed by release of the 3'-terminal fragment. Crystals of GreA have been obtained from polyethylene glycol 4000, 2-propanol and sodium citrate, pH 5.6 and have been propagated by a novel seeding procedure. The crystals diffract beyond 2 A resolution and belong to the orthorhombic space group P2(1)2(1)2(1), with cell dimensions a = 101.7 A, b = 42.22 A, c = 40.05 A and with one molecule in the asymmetric unit.

Bacterial Proteins↗

PubMed1994

GreA-induced transcript cleavage is accompanied by reverse translocation to a different transcription complex conformation.

GreA- and GreB-induced transcript cleavage drives reverse translocation of Escherichia coli RNA polymerase on a DNA template in the absence of NTPs (Feng, G.-H., Lee, D. N., Wang, D., Chan, C. L., and Landick, R. (1994) J. Biol. Chem. 269, 22282-22294, accompanying report). During transcript elongation, the sizes of the DNA footprint and the single-stranded transcription bubble vary markedly among transcription complexes halted at different template positions. To test whether transcription complex intermediates formed during transcript cleavage-induced reverse translocation also display heterogeneous conformations at different template positions, we examined the structures of two different transcription complexes before and after GreA treatment. Transcription complexes halted at position +16 after initiation at the T7 A1 promoter or paused at the trpL pause site exhibited strong blocks to transcript cleavage after removal of 6 to 10 nucleotides. In both cases, the down-stream contact between RNA polymerase and DNA moved little during transcript cleavage, thereby increasing its distance from the active site, whereas the upstream DNA contact and the borders of the transcription bubble moved in approximate register with the transcript 3'-end. The backward movements of halted E. coli RNA polymerase are similar to a recently postulated model for discontinuous translocation during transcription, but differ from those reported for arrested RNA polymerase II transcription complexes.

Bacterial Proteins↗

PubMed1994

The mutL repair gene of Escherichia coli K-12 forms a superoperon with a gene encoding a new cell-wall amidase.

We report a molecular genetic analysis of the region immediately upstream from the Escherichia coli mutL DNA repair gene at 94.8 min. An open reading frame ending 9 bp upstream from the start of mutL corresponds to a 48 kDa polypeptide detected previously in minicells. The predicted amino acid sequence of this 48 kDa polypeptide shows homology to the major N-acetylmuramoyl-L-alanine amidase autolysin of Bacillus subtilis, a known amidase of Bacillus licheniformis, and the product of a Salmonella typhimurium gene that maps near 50 min. Insertions in this upstream gene, which we named amiB, or in mutL did not affect cell shape or viability; however, overexpression of the AmiB polypeptide caused cell lysis, hypersensitivity to osmotic shock and treatment with water, and temporary autolysis by low levels of antibiotics, which are all consistent with AmiB acting as a cell-wall hydrolase. Analysis of chromosomal transcription demonstrated that amiB forms a complex operon with mutL and two additional upstream genes. mutL transcripts also originated from an internal promoter, designated PmutL, located in amiB 312 bp upstream from the translational start of mutL. Together, these results suggest that E. coli contains a second amidase possibly involved in cell-wall hydrolysis, septation, or recycling, and that transcription of this amidase is directly linked to a gene central for DNA repair.

Adenosine Triphosphatases↗

PubMed1994

[Clinicopathological analysis of rhabdomyosarcoma of the ocular adenexa].

12 cases including 11 males and 1 female of rhabdomyosarcoma of ocular adenexa were analysed. The average age of the patients was 4.2 years. 9 cases of lesion were located in the orbit, 2 in the eyelid and 1 in conjunctiva. Pathologically 8 cases were embryonal forms, 2 alveolar, 2 polymorphic. Immunohistochemical studies were performed in 8 cases, which showed desmin positive. Myoglobin presented strong or weak positive in 5 cases, suspicious positive in 3 cases. The clinical manifestation, prognosis and treatment were briefly discussed.

Child↗

PubMed1994

[Experimental study and clinical application of end-to-side (transverse incision) anastomosis between small arteries].

The results of end to side anastomoses with longitudinal or transverse incision on the transverse limb were investigated. The anastomosis of the superficial epigastric artery to the femoral artery in rabbits served as the experimental model. The results demonstrated that the microartery end to side anastomosis with transverse arteriotomy is a more preferable anastomosis style with higher patency, easier arteriotomy and suturing, and it effectively prevents anastomotic stoma spasm or stenosis, and in accord with vascular physiological function. It is worthy to be recommended. The clinical successful cases are also reported.

Abdominal Muscles↗

PubMed1993

The natural history of dry type of age-related macular degeneration.

To study the natural history of dry type of age-related macular degeneration (AMD) and search for a sensitive method for detecting the development of the disease, the fundus fluorescein angiography, visual acuity, electroretinogram and FM 100-hue test were used to examine 75 eyes, 147 eyes, 73 eyes, and 94 eyes respectively. These examinations were taken at least twice during the follow-up periods. The average age was 63.2 years (50-80 years). The average follow-up was 29.8 months with a range of 3-74 months. It was shown tht there were not any statistically significant difference in the macular lesions and electroretinogram between the initial examinations and after follow-up (P > 0.05). 91.14% of the eyes maintained good visual acuity during the follow-up. Subretinal neovascularization developed only in one of the eyes. The total error score of FM 100-hue test had a statistically significant difference between the initial test and the test taken two years afterwards (P < 0.01). It was suggested that most of the dry type of AMD had a favorable prognosis and that color visual test was a sensitive method for monitoring the development of dry type of AMD.

Age Factors↗

PubMed1993

Dynamic investigation on chromosome aberration of a human retinoblastoma cell line So-Rb50.

G-banding and karyotype analyses of cells in seventeen passages of SO-Rb50 during a long period of culture for about four years were performed. Three chromosome markers 13q14-, 1p36+ and 12p13+ were found. Cells possessed 13q14- reduced to zero after the 200th passage while 1p+ and 12p+ cells increased to 100% after 30 and 200 passages respectively. Abnormal chromosomes, ring chromosomes, chromosome radiuses and double minutes were also observed. These chromosomal changes were more often seen before the 200th passage. The significance of these changes are discussed.

Chromosome Aberrations↗

PubMed1993

[Pathologic, electron microscopic and experimental examination of band keratopathy].

One hundred and thirteen cases previously diagnosed as band keratopathy were reexamined by light microscopy and histochemistry, 10 cases of which were studied by TEM and 2 cases were studied by SEM. We find that the calcium deposition initially occur as fine calcific particles within the cytoplasm of epithelic cells and in the basal membrane and bowman's layer, which aggregated to form calcific spherules and large calcific masses extracellularly. When 3 normal corneal tissues were frozen and cultured, the calcium deposition occur in the superficial layer of bowman's membrane and in the stromal lamellae, Authors suggest that the degeneration and calcific abnormal metabolism of the cell caused by various factors is the main pathogenesis that leads to the calcific deposit. The evaporation and constructive destruction of the corneal surface in the interpiperbral area inhibits the formation of the lacrimal membrane and results in deposition of calcium salts in the superficial tissue of exposed cornea in a band configuration. The combination of glycosaminoglycan with calcium salts is the dynamics of aggregation of the fine calcific particles to form the calcific spherules and calcific masses.

Calcium↗