Biomedical subjects

D J Andrew

Publications and source records attributed to D J Andrew.

At least 19 recordsLinked to original sources

PubMed2001

pasilla, the Drosophila homologue of the human Nova-1 and Nova-2 proteins, is required for normal secretion in the salivary gland.

From a screen for genes expressed and required in the Drosophila salivary gland, we identified pasilla (ps), which encodes a set of proteins most similar to human Nova-1 and Nova-2. Nova-1 and Nova-2 are nuclear RNA-binding proteins normally expressed in the CNS where they directly regulate splicing. In patients suffering from paraneoplastic opsoclonus myoclonus ataxia (POMA), Nova-1 and Nova-2 proteins are present as auto-antigens. Consistent with a role in splicing, PS is localized to nuclear puncta. The salivary glands of ps mutants internalize normally and maintain epithelial polarity. However, the mutant salivary glands develop irregularities in overall morphology and have defects in apical secretion. The secretory defects in ps mutants provide a potential mechanism for the loss of motor function observed in POMA patients.

Alleles↗

PubMed2001

Organ formation in Drosophila: specification and morphogenesis of the salivary gland.

The Drosophila salivary gland has emerged as an outstanding model system for the process of organ formation. Many of the component steps, from initial regional specification through cell specialization and morphogenesis, are known and many of the genes required for these different processes have been identified. The salivary gland is a relatively simple organ; the entire gland comprises of only two major cell types, which derive from a single contiguous primordium. Salivary cells cease dividing once they are specified, and organ growth is achieved simply by an increase in size of individual cells, thus eliminating concerns about the potential unequal distribution of determinants during mitosis. Drosophila salivary glands form by the same cellular mechanisms as organs in higher organisms, including regulated cell shape changes, cell intercalation and directed cell migration. Thus, learning how these events are coordinated for tissue morphogenesis in an organism for which the genetic and molecular tools are unsurpassed should provide excellent paradigms for dissecting related processes in the more intricate organs of more complicated species.

Animals↗

PubMed2001

Regulation of Drosophila tracheal system development by protein kinase B.

Protein kinase B (PKB, also termed Akt) is a phosphatidylinositol 3' kinase (PI3'K)-dependent enzyme implicated in survival signaling and human tumorigenesis. To identify potential targets of this protein kinase, we employed a genetic screen in Drosophila. Among several genes that genetically interacted with PKB was trachealess (trh), which encodes a bHLH-PAS domain transcription factor required for development of the trachea and other tubular organs. Trh activates expression of the fibroblast growth factor receptor Breathless, which, in turn, is required for directed migration of all tracheal branches. Using a combination of biochemical and transgenic approaches, we show that direct phosphorylation of Trh by PKB at serine 665 is essential for nuclear localization and functional activation of this regulator of branching morphogenesis.

Active Transport, Cell Nucleus↗

PubMed2001

ribbon encodes a novel BTB/POZ protein required for directed cell migration in Drosophila melanogaster.

During development, directed cell migration is crucial for achieving proper shape and function of organs. One well-studied example is the embryonic development of the larval tracheal system of Drosophila, in which at least four signaling pathways coordinate cell migration to form an elaborate branched network essential for oxygen delivery throughout the larva. FGF signaling is required for guided migration of all tracheal branches, whereas the DPP, EGF receptor, and Wingless/WNT signaling pathways each mediate the formation of specific subsets of branches. Here, we characterize ribbon, which encodes a BTB/POZ-containing protein required for specific tracheal branch migration. In ribbon mutant tracheae, the dorsal trunk fails to form, and ventral branches are stunted; however, directed migrations of the dorsal and visceral branches are largely unaffected. The dorsal trunk also fails to form when FGF or Wingless/WNT signaling is lost, and we show that ribbon functions downstream of, or parallel to, these pathways to promote anterior-posterior migration. Directed cell migration of the salivary gland and dorsal epidermis are also affected in ribbon mutants, suggesting that conserved mechanisms may be employed to orient cell migrations in multiple tissues during development.

Amino Acid Sequence↗

PubMed2000

D-Titin: a giant protein with dual roles in chromosomes and muscles.

Previously, we reported that chromosomes contain a giant filamentous protein, which we identified as titin, a component of muscle sarcomeres. Here, we report the sequence of the entire titin gene in Drosophila melanogaster, D-Titin, and show that it encodes a two-megadalton protein with significant colinear homology to the NH(2)-terminal half of vertebrate titin. Mutations in D-Titin cause chromosome undercondensation, chromosome breakage, loss of diploidy, and premature sister chromatid separation. Additionally, D-Titin mutants have defects in myoblast fusion and muscle organization. The phenotypes of the D-Titin mutants suggest parallel roles for titin in both muscle and chromosome structure and elasticity, and provide new insight into chromosome structure.

Animals↗

PubMed2000

Salivary gland development in Drosophila melanogaster.

The Drosophila salivary gland is proving to be an excellent experimental system for understanding how cells commit to specific developmental programs and, once committed, how cells implement such decisions. Through genetic studies, the factors that determine where salivary glands will form, the number of cells committed to a salivary gland fate, and the distinction between the two major cell types (secretory cells and duct cells) have been discovered. Within the next few years, we will learn the molecular details of the interactions among the salivary gland regulators and salivary gland target genes. We will also learn how the early-expressed salivary gland genes coordinate their activities to mediate the morphogenetic movements required to form the salivary gland and the changes in cell physiology required for high secretory activity.

Animals↗

PubMed2000

Regulation and function of Scr, exd, and hth in the Drosophila salivary gland.

Salivary gland formation in the Drosophila embryo is dependent on the homeotic gene Sex combs reduced (Scr). When Scr function is missing, salivary glands do not form, and when SCR is expressed everywhere in the embryo, salivary glands form in new places. Scr is normally expressed in all the cells that form the salivary gland. However, as the salivary gland invaginates, Scr mRNA and protein disappear. Homeotic genes, such as Scr, specify tissue identity by regulating the expression of downstream target genes. For many homeotic proteins, target gene specificity is achieved by cooperatively binding DNA with cofactors. Therefore, it is likely that SCR also requires a cofactor(s) to specifically bind to DNA and regulate salivary gland target gene expression. Here, we show that two homeodomain-containing proteins encoded by the extradenticle (exd) and homothorax (hth) genes are also required for salivary gland formation. exd and hth function at two levels: (1) exd and hth are required to maintain the expression of Scr in the salivary gland primordia prior to invagination and (2) exd and hth are required in parallel with Scr to regulate the expression of downstream salivary gland genes. We also show that Scr regulates the nuclear localization of EXD in the salivary gland primordia through repression of homothorax (hth) expression, linking the regulation of Scr activity to the disappearance of Scr expression in invaginating salivary glands.

Animals↗

PubMed2000

Titin as a chromosomal protein.

We identified titin as a chromosomal protein using a human autoimmune scleroderma serum. We cloned the corresponding gene in the fruitfly, Drosophila melanogaster. We have demonstrated that titin is not only expressed and localized in striated muscle but is also distributed uniformly on condensed mitotic chromosomes using multiple antibodies directed against different domains of both Drosophila and vertebrate titin. Titin is a giant sarcomeric protein responsible for the elasticity of striated muscle. Titin may also function as a molecular scaffold during myofibril assembly. We hypothesize that titin is a component of chromosomes that may function to determine chromosome structure and provide elasticity, playing a role similar to that proposed for titin in muscle. We have identified mutations in Drosophila Titin (D-Titin) and are characterizing phenotypes in muscle and chromosomes.

Animals↗

PubMed2000

Early genes required for salivary gland fate determination and morphogenesis in Drosophila melanogaster.

Studies of Drosophila salivary gland formation have elucidated the regulatory pathway by which the salivary gland fate is determined and the morphogenetic processes by which the primordial cells are internalized to form the tubular glands. Both the position of the salivary primordia and the number of cells recruited to a salivary gland fate are established through a combination of the localized expression of the transcription factors SEX COMBS REDUCED (SCR), TEASHIRT (TSH) and ABDOMINAL-B (ABD-B), and localized DPP-signaling. Similarly, the distinction between the two major cell types, duct and secretory, is determined by spatially limited EGF-signaling. Salivary gland formation also requires the function of two transcription factors expressed in nearly all cells of the developing embryo, EXTRADENTICLE (EXD) and HOMOTHORAX (HTH). Once the salivary gland fate is determined, cells of the secretory primordia are internalized by an apical constriction mode of invagination. We have characterized three genes encoding transcription factors, trachealess (trh), hückebein (hkb), and fork head (fkh), that are downstream targets of the salivary gland regulators. Mutations in these transcription factors profoundly affect salivary gland morphogenesis. trh is required for the formation of the salivary duct tubes. hkb determines the order of secretory cell invagination, a regulated process critical for determining the final shape of the salivary gland. fkh has two early roles in salivary gland formation. fkh both promotes secretory cell survival and facilitates secretory cell internalization. trh, hkb, and fkh are involved in the formation of not only the salivary duct and secretory tubes, but also of other tubular structures, such as the trachea and the gut endoderm. We propose that trh, hkb, and fkh may serve as "morphogenetic cassettes" responsible for forming tubular structures in a variety of tissues.

Animals↗

PubMed2000

Fork head prevents apoptosis and promotes cell shape change during formation of the Drosophila salivary glands.

The secretory tubes of the Drosophila salivary glands are formed by the regulated, sequential internalization of the primordia. Secretory cell invagination occurs by a change in cell shape that includes basal nuclear migration and apical membrane constriction. In embryos mutant for fork head (fkh), which encodes a transcription factor homologous to mammalian hepatocyte nuclear factor 3beta (HNF-3beta), the secretory primordia are not internalized and secretory tubes do not form. Here, we show that secretory cells of fkh mutant embryos undergo extensive apoptotic cell death following the elevated expression of the apoptotic activator genes, reaper and head involution defective. We rescue the secretory cell death in the fkh mutants and show that the rescued cells still do not invaginate. The rescued fkh secretory cells undergo basal nuclear migration in the same spatial and temporal pattern as in wild-type secretory cells, but do not constrict their apical surface membranes. Our findings suggest at least two roles for fkh in formation of the embryonic salivary glands: an early role in promoting survival of the secretory cells, and a later role in secretory cell invagination, specifically in the constriction of the apical surface membrane.

Animals↗

PubMed2000

Organ shape in the Drosophila salivary gland is controlled by regulated, sequential internalization of the primordia.

During Drosophila development, the salivary primordia are internalized to form the salivary gland tubes. By analyzing immuno-stained histological sections and scanning electron micrographs of multiple stages of salivary gland development, we show that internalization occurs in a defined series of steps, involves coordinated cell shape changes and begins with the dorsal-posterior cells of the primordia. The ordered pattern of internalization is critical for the final shape of the salivary gland. In embryos mutant for hückebein (hkb), which encodes a transcription factor, or faint sausage (fas), which encodes a cell adhesion molecule, internalization begins in the center of the primordia, and completely aberrant tubes are formed. The sequential expression of hkb in selected cells of the primordia presages the sequence of cell movements. We propose that hkb dictates the initial site of internalization, the order in which invagination progresses and, consequently, the final shape of the organ. We propose that fas is required for hkb-dependent signaling events that coordinate internalization.

Animals↗

PubMed1999

Cell fate specification in the Drosophila salivary gland: the integration of homeotic gene function with the DPP signaling cascade.

Salivary gland formation in the Drosophila embryo is linked to the expression of the homeotic gene Sex combs reduced (Scr). When Scr function is missing, salivary glands do not form, and when SCR is expressed everywhere, salivary glands form in new places. However, not every cell that expresses Scr is recruited to a salivary gland fate. Along the anterior-posterior axis, the posteriorly expressed proteins encoded by the teashirt (tsh) and Abdominal-B (Abd-B) genes block SCR activation of salivary gland genes, and along the dorsal-ventral axis, the secreted signaling molecule encoded by decapentaplegic (dpp) prevents activation of salivary gland genes by SCR in dorsal regions of parasegment 2. We have identified five downstream components in the DPP signaling cascade required to block salivary gland gene activation. These components include two known receptors, the type I receptor encoded by the thick veins (tkv) gene and the type II receptor encoded by the punt (put) gene; two of the four known Drosophila members of the Smad family of proteins which transduce signals from the receptors to the nucleus, Mothers against dpp (Mad) and Medea (Med); and, finally, a large zinc-finger transcription factor encoded by the schnurri (shn) gene. These results reveal how anterior-posterior and dorsal-ventral patterning information is integrated at the level of organ-specific gene expression.

Animals↗

PubMed1999

Aurintricarboxylic acid inhibits apoptosis and supports proliferation in a haemopoietic growth-factor dependent myeloid cell line.

The actions of the nuclease inhibitor aurintricarboxylic acid (ATA) were investigated in the growth-factor dependent murine myeloid cell line NSF-60. NSF-60 cells proliferate in response to interleukin-3 (IL-3) and undergo apoptosis when deprived of exogenous IL-3, as demonstrated by the appearance of characteristic DNA 'ladders' following agarose gel electrophoresis. ATA, at concentrations between 5 and 25 microM, inhibited apoptosis in growth-factor deprived cells as demonstrated by inhibition of DNA fragmentation and increased cell survival. ATA at a concentration of 25 microM supported proliferation of the cell line in the absence of exogenous growth-factor. Both ATA and IL-3 increased protein phosphorylation in this cell line. ATA and IL-3 induced proliferation was inhibited by the kinase inhibitors genistein, staurosporine and H-7. These findings suggest that, in NSF-60, ATA is not acting exclusively as an endonuclease inhibitor and that protein phosphorylation is involved in the mechanism of action of ATA in this cell line.

Animals↗

PubMed1999

WRS-85D: A tryptophanyl-tRNA synthetase expressed to high levels in the developing Drosophila salivary gland.

In a screen for genes expressed in the Drosophila embryonic salivary gland, we identified a tryptophanyl-tRNA synthetase gene that maps to cytological position 85D (WRS-85D). WRS-85D expression is dependent on the homeotic gene Sex combs reduced (Scr). In the absence of Scr function, WRS-85D expression is lost in the salivary gland primordia; conversely, ectopic expression of Scr results in expression of WRS-85D in new locations. Despite the fact that WRS-85D is a housekeeping gene essential for protein synthesis, we detected both WRS-85D mRNA and protein at elevated levels in the developing salivary gland. WRS-85D is required for embryonic survival; embryos lacking the maternal contribution were unrecoverable, whereas larvae lacking the zygotic component died during the third instar larval stage. We showed that recombinant WRS-85D protein specifically charges tRNATrp, and WRS-85D is likely to be the only tryptophanyl-tRNA synthetase gene in Drosophila. We characterized the expression patterns of all 20 aminoacyl-tRNA synthetases and found that of the four aminoacyl-tRNA synthetase genes expressed at elevated levels in the salivary gland primordia, WRS-85D is expressed at the highest level throughout embryogenesis. We also discuss the potential noncanonical activities of tryptophanyl-tRNA synthetase in immune response and regulation of cell growth.

Amino Acid Sequence↗

PubMed1998

Identification of a novel Drosophila SMAD on the X chromosome.

TGF-beta signaling from the cell surface to the nucleus is mediated by the SMAD family of proteins, which have been grouped into three classes based upon sequence identity and function. Receptor-regulated, or pathway-restricted, SMADs (R-SMADs) are phosphorylated by ligand-specific serine/threonine kinase receptors. Phosphorylated R-SMADs oligomerize with the coactivating, or shared, SMAD (Co-SMAD) mediator and translocate to the nucleus where the complex directs transcription of downstream target genes. Inhibitory SMADs (I-SMADs) block receptor-mediated phosphorylation of R-SMADs. In Drosophila, one member of each class of SMAD has been reported: MAD, an R-SMAD, MEDEA, a Co-SMAD, and DAD, an I-SMAD. Here, we report the first identification of a novel Drosophila R-SMAD, which we have named Smox for Smad on X. We have localized the Smox gene to a specific interval on the X chromosome and shown that Smox is transcribed throughout development.

Amino Acid Sequence↗

PubMed1998

Human autoantibodies reveal titin as a chromosomal protein.

Assembly of the higher-order structure of mitotic chromosomes is a prerequisite for proper chromosome condensation, segregation and integrity. Understanding the details of this process has been limited because very few proteins involved in the assembly of chromosome structure have been discovered. Using a human autoimmune scleroderma serum that identifies a chromosomal protein in human cells and Drosophila embryos, we cloned the corresponding Drosophila gene that encodes the homologue of vertebrate titin based on protein size, sequence similarity, developmental expression and subcellular localization. Titin is a giant sarcomeric protein responsible for the elasticity of striated muscle that may also function as a molecular scaffold for myofibrillar assembly. Molecular analysis and immunostaining with antibodies to multiple titin epitopes indicates that the chromosomal and muscle forms of titin may vary in their NH2 termini. The identification of titin as a chromosomal component provides a molecular basis for chromosome structure and elasticity.

Amino Acid Sequence↗

PubMed1998

Regulation and formation of the Drosophila salivary glands.

The homeotic gene, Sex combs reduced (Scr), is a master regulator of Drosophila salivary gland formation. Embryos in which Scr function is missing do not form salivary glands, and embryos in which SCR protein is expressed everywhere form extra salivary glands. However, other known proteins, including the homeotic protein Abdominal-B, the unusual zinc finger protein Teashirt, and the secreted signaling molecule Decapentaplegic (a TGF-beta family member), limit the recruitment of SCR-expressing cells to salivary glands. To learn the molecular details of how salivary gland gene expression is controlled and as a first step toward understanding how the SCR transcription factor controls salivary gland morphogenesis, we screened for genes expressed in the developing salivary gland. Among our best candidates for potential direct downstream targets of SCR in the salivary gland are the genes trachealess (trh), dCREB-A, jalapeño, and Semaphorin II (SemaII). Our genetic studies suggest distinct and important roles for each of these genes in salivary gland morphogenesis. Current work includes studying the molecular interactions between SCR and these downstream target genes and asking how target genes coordinate their activities to effect the cell biological changes required to build functional salivary glands.

Animals↗

PubMed1998

Protection of human upper respiratory tract cell lines against sulphur mustard toxicity by hexamethylenetetramine (HMT).

1. Sulphur mustard ('mustard gas', HD) is a highly toxic chemical warfare agent which affects the skin and respiratory tract. The primary targets of inhaled HD are the epithelia of the upper respiratory tract. Hexamethylenetetramine (HMT) has been shown to protect human lung cells against HD toxicity and has also been shown to be effective in vivo against the chemical warfare agent phosgene. The ability of HMT to protect against the toxicity of HD was investigated in the human upper respiratory tract cell lines BEAS-2B and RPMI 2650. 2. HD was highly toxic to both cell lines, with LC50 values of 15-30 microM. HMT, at a concentration of 10 mM, was shown to protect the cell lines against the toxic effects of 20 microM and 40 microM HD. Results demonstrated that it was necessary for HMT to be in situ at the time of exposure to HD for effective cytoprotection. No protection was seen when cells were treated with HMT following exposure to HD, or where HMT was removed prior to HD exposure. 3. Results suggest that HMT may be effective prophylaxis for exposure to HD by inhalation.

Anti-Infective Agents, Urinary↗