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D Yamamoto

Publications and source records attributed to D Yamamoto.

At least 73 records · Page 4Linked to original sources

Regulation of cone cell formation by Canoe and Ras in the developing Drosophila eye.

Cone cells are lens-secreting cells in ommatidia, the unit eyes that compose the compound eye of Drosophila. Each ommatidium contains four cone cells derived from precursor cells of the R7 equivalence group which express the gene sevenless (sev). When a constitutively active form of Ras1 (Ras1V12) is expressed in the R7 equivalence group cells using the sev promoter (sev-Ras1V12), additional cone cells are formed in the ommatidium. Expression of Ras1N17, a dominant negative form of Ras1, results in the formation of 1-3 fewer cone cells than normal in the ommatidium. The effects of Ras1 variants on cone cell formation are modulated by changing the gene dosage at the canoe (cno) locus, which encodes a cytoplasmic protein with Ras-binding activity. An increase or decrease in gene dosage potentiates the sev-Ras1v12 action, leading to marked induction of cone cells. A decrease in cno+ activity also enhances the sev-Ras1N17 action, resulting in a further decrease in the number of cone cells contained in the ommatidium. In the absence of expression of sev-Ras1V12 or sev-Ras1N17, an overdose of wild-type cno (cno+) promotes cone cell formation while a significant reduction in cno+ activity results in the formation of 1-3 fewer cone cells than normal in the ommatidium. We propose that there are two signaling pathways in cone cell development, one for its promotion and the other for its repression, and Cno functions as a negative regulator for both pathways. We also postulate that Cno predominantly acts on a prevailing pathway in a given developmental context, thereby resulting in either an increase or a decrease in the number of cone cells per ommatidium. The extra cone cells resulting from the interplay of Ras1v12 and Cno are generated from a pool of undifferentiated cells that are normally fated to develop into pigment cells or undergo apoptosis.

Animals↗

The Drosophila mushroom body is a quadruple structure of clonal units each of which contains a virtually identical set of neurones and glial cells.

The mushroom body (MB) is an important centre for higher order sensory integration and learning in insects. To analyse the development and organisation of the MB neuropile in Drosophila, we performed cell lineage analysis in the adult brain with a new technique that combines the Flippase (flp)/FRT system and the GAL4/UAS system. We showed that the four mushroom body neuroblasts (MBNbs) give birth exclusively to the neurones and glial cells of the MB, and that each of the four MBNb clones contributes to the entire MB structure. The expression patterns of 19 GAL4 enhancer-trap strains that mark various subsets of MB cells revealed overlapping cell types in all four of the MBNb lineages. Partial ablation of MBNbs using hydroxyurea showed that each of the four neuroblasts autonomously generates the entire repertoire of the known MB substructures.

Animals↗

Molecular cloning and characterization of the ABC transporter expressed in Trachea (ATET) gene from Drosophila melanogaster.

A novel member of the ATP-binding cassette (ABC) transporter proteins has been cloned from Drosophila melanogaster. The gene is designated as ABC Transporter Expressed in Trachea (Atet), because the transcript was localized to the respiratory system by in situ hybridization analysis of whole-mount embryos using digoxigenin-labeled RNA probes. The hybridization signal was also observed in amnioserosa. Northern blot analysis identified a single 4.5 kb mRNA expressed in all developmental stages at a relatively constant level. The Atet gene mapped to 24E on the left arm of the second chromosome. The Atet protein shows extensive homology with the Drosophila white gene product, which is reported to form heterodimers with the brown or scarlet gene product to transport guanine or tryptophan into the pigment cells in the compound eye. By analogy, Atet is suggested to be involved in transporting a small molecule after dimerization with a partner protein.

ATP-Binding Cassette Transporters↗

Sexual orientation in Drosophila is altered by the satori mutation in the sex-determination gene fruitless that encodes a zinc finger protein with a BTB domain.

We have isolated a new Drosophila mutant, satori (sat), the males of which do not court or copulate with female flies. The sat mutation comaps with fruitless (fru) at 91B and does not rescue the bisexual phenotype of fru, indicating that sat is allelic to fru (fru(sat)). The fru(sat) adult males lack a male-specific muscle, the muscle of Lawrence, as do adult males with other fru alleles. Molecular cloning and analyses of the genomic and complementary DNAs indicated that transcription of the fru locus yields several different transcripts. The sequence of fru cDNA clones revealed a long open reading frame that potentially encodes a putative transcription regulator with a BTB domain and two zinc finger motifs. In the 5' noncoding region, three putative transformer binding sites were identified in the female transcript but not in male transcripts. The fru gene is expressed in a population of brain cells, including those in the antennal lobe, that have been suggested to be involved in determination of male sexual orientation. We suggest that fru functions downstream of tra in the sex-determination cascade in some neural cells and that inappropriate sexual development of these cells in the fru mutants results in altered sexual orientation of the fly.

Alleles↗

Identification of AF-6 and canoe as putative targets for Ras.

Ras (Ha-Ras, Ki-Ras, N-Ras) is implicated in the regulation of various cell functions such as gene expression and cell proliferation downstream from specific extracellular signals. Here, we partially purified a Ras-interacting protein with molecular mass of about 180 kDa (p180) from bovine brain membrane extract by glutathione S-transferase (GST)-Ha-Ras affinity column chromatography. This protein bound to the GTP gamma S (guanosine 5'-(3-O-thio)triphosphate, a nonhydrolyzable GTP analog).GST-Ha-Ras affinity column but not to those containing GDP.GST-Ha-Ras or GTP gamma S.GST-Ha-Ras with a mutation in the effector domain (Ha-RasA38). The amino acid sequences of the peptides derived from p180 were almost identical to those of human AF-6 that is identified as the fusion partner of the ALL-1 protein. The ALL-1/AF-6 chimeric protein is the critical product of the t (6:11) abnormality associated with some human leukemia. AF-6 has a GLGF/Dlg homology repeat (DHR) motif and shows a high degree of sequence similarity with Drosophila Canoe, which is assumed to function downstream from Notch in a common developmental pathway. The recombinant N-terminal domain of AF-6 and Canoe specifically interacted with GTP gamma S.GST-Ha-Ras. The known Ras target c-Raf-1 inhibited the interaction of AF-6 with GTP gamma S.GST-Ha-Ras. These results indicate that AF-6 and Canoe are putative targets for Ras.

Amino Acid Sequence↗

Purification and characterization of angiotensin II-generating chymase from hamster cheek pouch.

Hamster cheek pouch vascular tissues contain an angiotensin II-forming enzyme which is inhibited by chymostatin but not by any angiotensin-converting enzyme inhibitors. The enzyme was purified to apparent homogeneity by gel filtration and heparin-Sepharose affinity chromatography. The molecular mass estimated by sodium dodecyl sulphate polyacrylamide gel electrophoresis was 28 kDa and the optimum pH was between 7.5 and 9.0. The angiotensin II-forming activity was inhibited by chymostatin, soybean trypsin inhibitor and phenylmethylsulfonyl fluoride, but not by aprotinin. The N-terminal sequence showed high homology with chymases from various species. Thus, the angiotensin II-generating enzyme obtained from hamster cheek pouch vessels is a chymase.

Amino Acid Sequence↗

A nonpeptidal peptidomimetic agonist of the insect FLRFamide myosuppressin family.

Benzethonium chloride (Bztc) is the first totally nonpeptide ligand for an insect, indeed an invertebrate, peptide receptor. Bztc mimics the inhibitory physiological activity of the myosuppressins, a subfamily of the FLRFamides, in three different insect bioassay systems. The inhibitory action of leucomyosuppressin and the nonpeptide Bztc in both the cockroach hindgut and the mealworm neuromuscular junction can be blocked by the lipoxygenase inhibitor, nordihydroguaiaretic acid, providing evidence for similar modes of action. Lipoxygenase metabolites of arachidonic acid may mediate inhibition of neuromuscular transmission by these two factors. In addition, Bztc competitively displaces a radiolabeled myosuppressin analogue from high- and low-affinity receptors of the locust oviduct. Thus, the nonpeptide interacts with both binding and activating regions of myosuppressin receptors. Molecular dynamics experiments in which selected functional groups of Bztc were fit onto corresponding functional groups of low-energy myosuppressin pentapeptide structures indicate how Bztc may mimic the myosuppressins at a molecular level. The discovery of Bztc as a nonpeptidal peptidomimetic analogue provides an opportunity to develop new pest management strategies by targeting an insect's own peptide receptor.

Amino Acid Sequence↗

Genetic dissection of sexual orientation: behavioral, cellular, and molecular approaches in Drosophila melanogaster.

Insertional mutagenesis using P-element vectors yielded several independent mutations that cause male homosexuality in Drosophila melanogaster. Subsequent analyses revealed that all of these insertions were located at the same chromosomal division, 91B, where one of the inversion breakpoints responsible for the bisexual phenotype of the fruitless (fru) mutant has been mapped. In addition to the altered sexual orientation, the fru mutants displayed a range of defects in the formation of a male-specific muscle, the muscle of Lawrence. Since the male-specific formation of this muscle was dependent solely on the sex of the innervating nerve and not on the sex of the muscle itself, the primary site of action of the fru gene should be in the neural cells. satori, one of the P-insertion alleles of fru which we isolated, carried the lacZ gene of E. coli as a reporter, and beta-galactosidase expression was found in a subset of brain cells including those in the antennal lobe in the satori mutant. Targeted expression of a sex determination gene, transformer (tra), was used to produce chromosomally male flies with certain feminized glomeruli in the antennal lobe. Such sexually mosaic flies courted not only females but also males when the DM2, DA3 and DA4 glomeruli were feminized, indicating that these substructures in the antennal lobe may be involved in the determination of the sexual orientation of flies. Molecular cloning and analyses of the genomic and complementary DNAs indicated that transcription of the fru locus yields several different transcripts, one of which encodes a putative transcription regulator with a BTB domain and two zinc finger motifs. In the 5' non-coding region, three putative Transformer binding sites were identified. It appears plausible therefore that the fru gene is one of the elements in the sex determination cascade that controls sexual fates of certain neuronal cells. Improper sex determination in these neural cells may lead to altered sexual orientation and malformation of the male-specific muscle. Some implications of the results of our study on sexual orientation in other organisms will be discussed based on the Drosophila research.

Amino Acid Sequence↗

canoe encodes a novel protein containing a GLGF/DHR motif and functions with Notch and scabrous in common developmental pathways in Drosophila.

The canoemisty1 (cnomis1) mutation was isolated by virtue of its severe rough eye phenotype from approximately 500 fly lines, each harboring a single autosomal insertion of a P element (Bm delta w). Excision of the P element generated a lethal, null allele, cnomis10, together with many revertants with normal eye morphology. Ommatidia homozygous for cnomis10, produced in an otherwise wild-type eye by somatic recombination, typically contain a reduced number of outer photoreceptors. Some cnomis1 homozygous adults bear extra macrochaetes on the head, notum, humerus and/or scutellum. cnomis1 hemizygotes often show conspicuous wing phenotypes such as a notched blade and the loss of a cross vein. The sequence of cno cDNA clones isolated from an embryonic cDNA library revealed a long open reading frame that potentially encodes a 1893-amino-acid protein with the GLGF/DHR motif, a conserved sequence in Discs large, Dishevelled, and some other proteins associated with cellular junctions. Flies doubly mutant for cnomis1 and scabrous1 (sca1) and those for cnomis1 and the split (spl) allele of Notch (N) always have rumpled wings curved downward. The spl; cnomis1 double mutant flies also exhibit a "giant socket" phenotype. These phenotypes are rarely observed flies singly mutant for either cnomis1, sca1 or spl. The wing vein gaps caused by Abruptex1, a N allele producing an activated form of N protein, are dominantly suppressed by cnomis1. Heterozygosity for shaggy and myospheroid promotes formation of extra wing veins in cnomis1 homozygotes. The genetic interactions suggest that cno participates with members of the N pathway in regulating adhesive cell-cell interactions for the determination of cell fate.

Amino Acid Sequence↗

Diagnostic relevance of abortion-associated human embryonic antigen expressed on the cell surface of tumour promoter-treated Bloom syndrome cells.

We detected stable expression of human embryonic antigen associated with spontaneous abortion (HEAA) on the cell surface of a tumour promoter-treated B lymphoblastoid cell line (BS-SHY) originating from Bloom syndrome. We used indirect immunofluorescence and diluted serum from 44 patients who had recurrent spontaneous abortions. With the use of the panning procedure, we separated characteristic cells expressing strong HEAA. The BS-SHY-HEAA cells separated here would be useful for measuring serum antibody (against HEAA) produced by patients with recurrent abortions. It was also noted that aborters who received husbands' leukocyte immunization have lost this antibody, and have delivered successful pregnancies at term. Using HEAA proteins, we conducted Western blotting analysis for the amino acid sequencing (mol. wt 77 kDa). Amino acid sequencing data indicated that HEAA had 87.5% homology to the immunoglobulin (Ig) VHIII region in the framework. Recently, the protective value of high dose i.v. administration of immunoglobulin in the treatment of recurrent spontaneous abortions has been reported to be similar to that of leukocyte immunization. Therefore, the BS-SHY-HEAA cells appear to provide a valuable tool for rapid serological diagnosis and for evaluating the efficacy of immunotherapy with husbands' leukocytes in patients with recurrent spontaneous abortions.

Abortion, Habitual↗

Phenotypic and molecular characterization of croaker, a new mating behavior mutant of Drosophila melanogaster.

Mating of Drosophila melanogaster is a stereotypically patterned behavior consisting of a fixed sequence of actions that is primarily under genetic control. Although courtship can be easily monitored and quantified, little is known about its neural basis. To obtain a better understanding of cellular and molecular mechanisms underlying courtship, we have isolated mutants that disrupt specific aspects of mating behavior. The croaker mutant was isolated from approximately 1,000 lines harboring single P-element insertions by screening for aberrant courtship song: croaker males often generate polycyclic pulse song while most of the song pulses are monocyclic in the wild-type. The mutant is also defective in flight. Intracellular recordings of excitatory junction potentials from larval body wall muscles and Ca++ action potentials from adult indirect flight muscles demonstrated that neuromuscular transmission and Ca++ electrogenesis in the muscle fibers are not impaired by the croaker mutation. To define the croaker gene molecularly, genomic DNA surrounding the P-element insertion site was cloned by plasmid rescue and subsequent screening of a cosmid library. Northern blotting with the genomic DNA probes detected three transcripts in the wild-type, which were not expressed in the croaker mutant.

Animals↗

Signaling mechanisms in induction of the R7 photoreceptor in the developing Drosophila retina.

The Drosophila compound eye is an excellent experimental system for analysing fate induction of identifiable single cells. Each ommatidium, a unit eye, contains eight photoreceptors (R1-R8), and the differentiation of these photoreceptors occurs in the larval eye imaginal disc in discrete steps: first R8 is determined, then R2/R5, R3/R4, R1/R6 and finally R7. Induction of R7, in particular, has been extensively studied at the molecular level. The R8 photoreceptor presents on its surface a ligand, Bride of Sevenless, that binds and activates Sevenless receptor tyrosine kinase in the R7 precursor. Autophosphorylated Sevenless initiates a Ras1-mediated cascade, which eventually activates transcription factors in the nucleus via Raf1 and MAP kinases, resulting in R7 development. However, recent studies indicate that Sevenless (Sev) functions just to neuralize the cell and has no role in R7 fate determination per se. It appears that the R7 fate may represent the lowest rung of a 'neuronal ground state', which is attained without any specific inductive cue. It is plausible that the R7 precursor is actively prevented from taking on the neuronal fate and this inhibition is removed by activation of Sev.

Animals↗

Sequence and expression of a gene encoding a ribosomal protein S4 homolog from Drosophila melanogaster.

We describe a Drosophila melanogaster cDNA clone encoding an amino acid (aa) sequence 75% identical to human ribosomal protein S4 (RPS4). The D. melanogaster RPS4 has 260 aa, if the NH2-terminal methionine is removed after translation of the mRNA. The mRNA for the protein is about 1 kb in length and is detected throughout the developmental stages tested (i.e., embryo, larva, pupa and adult). The cDNA clone hybridizes to two sites on the X chromosome, 1B1-2 and 3A3-6.

Amino Acid Sequence↗

Positive and negative signaling mechanisms in the regulation of photoreceptor induction in the developing Drosophila retina. Review.

An ommatidium of a Drosophila compound eye contains eight photoreceptor cells, R1-R8. The fates of the photoreceptors are determined exclusively by inductive interactions between neuronal precursors in the cell cluster from which the ommatidium is formed. R7 induction has been extensively analysed at the molecular level. Activation of a membrane receptor tyrosine kinase (Sevenless) in the R7 precursor by a ligand (Bride of sevenless) present on the surface of R8 triggers a transduction cascade mediated by Ras, establishing the R7 fate of this cell. Other Sev-expressing cells are prevented from taking on the R7 fate by several different mechanisms. Pokkuri-mediated repression represents one such regulatory mechanism. The positive and negative signaling pathways operating in the fate determination of other photoreceptor cells are also discussed.

Animals↗

Effects of epidermal growth factor on ornithine decarboxylase activity and DNA synthesis in rats during the perinatal period.

The effects of epidermal growth factor (EGF) on ornithine decarboxylase (ODC) activity and DNA synthesis were studied during the perinatal period in rats. EGF administration to neonatal rats increased ODC activity and DNA synthesis in neonatal rat liver, but not in brain. EGF administration to maternal rats increased ODC activity in maternal rat liver, but not in rat placenta and fetal rat liver. These data suggest that EGF has mitogenic effects on neonatal and maternal rat liver and play some important roles in fetal and neonatal growth.

Animals↗

Crystal structure of papain-succinyl-Gln-Val-Val-Ala-Ala-p-nitroanilide complex at 1.7-A resolution: noncovalent binding mode of a common sequence of endogenous thiol protease inhibitors.

Succinyl-Gln-Val-Val-Ala-Ala-p-nitroanilide corresponding to a common sequence of endogenous thiol protease inhibitors is a noncompetitive reversible inhibitor of papain. In order to elucidate the binding mode of the inhibitor at the atomic level, its complex with papain was crystallized at ca. pH 7.0 using the hanging drop method, and the crystal structure was analyzed at 1.7-A resolution. The crystal has space group P2(1)2(1)2(1), with a = 43.09, b = 102.32, c = 49.69 A, and Z = 4. A total of 47,215 observed reflections were collected on the imaging plates using the same single crystal, and 19,833 unique reflections with Fo > sigma (Fo) were used for structure determination and refinement. The papain structure was determined by use of the atomic coordinates of papain previously reported, and then refined by the X-PLOR program. The inhibitor molecule was located on a difference Fourier map and fitted into the electron density with the aid of computer graphics. The complex structure was finally refined to R = 19.6% including 118 solvent molecules. The X-ray analysis of the complex crystal shows that the inhibitor is located at the R-domain side, not in the center of the binding site created by the R- and L-domains of papain. Such a binding mode of the inhibitor explains well the biological behavior that the inhibitor exhibits against papain. Comparison with the structure of papain-stefin B complex indicates that the structure of the Gln-Val-Val-Ala-Gly sequence itself is not necessarily the essential requisite for inhibitory activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Crystal structure of papain-E64-c complex. Binding diversity of E64-c to papain S2 and S3 subsites.

In order to investigate the binding mode of E64-c (a synthetic cysteine proteinase inhibitor) to papain at the atomic level, the crystal structure of the complex was analysed by X-ray diffraction at 1.9 A (1 A is expressed in SI units as 0.1 nm) resolution. The crystal has a space group P2(1)2(1)2(1) with a = 43.37, b = 102.34 and c = 49.95 A. A total of 21,135 observed reflections were collected from the same crystal, and 14811 unique reflections of up to 1.9 A resolution [Fo > 3 sigma(Fo)] were used for the structure solution and refinement. The papain structure was determined by means of the molecular replacement method, and then the inhibitor was observed on a (2 magnitude of Fo-magnitude of Fc) difference Fourier map. The complex structure was finally refined to R = 19.4% including 207 solvent molecules. Although this complex crystal (Form II) was polymorphous as compared with the previously analysed one (Form I), the binding modes of leucine and isoamylamide moieties of E64-c were significantly different from each other. By the calculation of accessible surface area for each complex atom, these two different binding modes were both shown to be tight enough to prevent the access of solvent molecules to the papain active site. With respect to the E64-c-papain binding mode, molecular-dynamics simulations proposed two kinds of stationary states which were derived from the crystal structures of Forms I and II. One of these, which corresponds to the binding mode simulated from Form I, was essentially the same as that observed in the crystal structure, and the other was somewhat different from the crystal structure of Form II, especially with respect to the binding of the isoamylamide moiety with the papain S subsites. The substrate specificity for the papain active site is discussed on the basis of the present results.

Amino Acid Sequence↗