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Homologs of the mouse Brachyury gene are involved in the specification of posterior terminal structures in Drosophila, Tribolium, and Locusta.

The Brachyury (T) gene is required for notochord differentiation in vertebrates. We have identified a Drosophila gene, the T-related gene (Trg), with high similarity to T within a stretch of approximately 200 amino acids, the DNA-binding domain of T. Trg is expressed throughout embryogenesis, first at the blastoderm stage in the hindgut primordium under the control of the terminal gap genes tll and hkb, and then until the end of embryogenesis in the differentiating hindgut. Drosophila embryos deficient for Trg do not form the hindgut, a phenotype that can be rescued by a Trg transgene. Thus, a common feature of T and Trg is their requirement in specifying the development of a single embryonic structure. Homologs of Trg are also expressed in the developing hindgut of Tribolium and Locusta embryos suggesting a highly conserved function of Trg in insects. This conservation and the high similarity of T and Trg raise the question of a common evolutionary origin of the hindgut of insects and the notochord of chordates.

Amino Acid Sequence↗

Ethylene dibromide: toxicology and risk assessment.

Since the 1920s ethylene dibromide's (EDB's) primary use has been as a scavenger of lead compounds in gasoline. Gasoline evaporation contributed to EDB emissions into the environment. In 1973, the United States Environmental Protection Agency (EPA) issued regulations to reduce the use of leaded gasoline and this has resulted in lower EDB usage and emissions. In addition, EDB has been used extensively as a fumigant since 1948. Its volatility and versatility, based on chemical and biocidal properties, led to its use as a soil sterilant, as a spot fumigant of grain milling machinery, and as a control agent in grain, fruit and vegetable infestations. In 1977 the EPA began a review of EDB's pesticidal uses which eventually led to its cancellation for most agricultural applications. Disposal of EDB and contamination of water supplies remain major environmental concerns. EDB can be absorbed via the dermal, oral and inhalation routes. It appears to be metabolized in vivo by an oxidative pathway (cytochrome P-450) and a conjugation pathway (glutathione S-transferase). The metabolites play an important role in exerting its toxicity. Few human poisonings have been reported from either acute or chronic exposure. However, EDB is irritating to the skin and eyes. Limited information indicates that EDB can damage the liver and kidneys following extensive or prolonged exposure. The genotoxicity of EDB has been clearly demonstrated. It binds to DNA in vivo and in vitro, and a DNA adduct has been identified. EDB has been shown to be mutagenic in numerous bacterial assays, in fungi, in plants, in insects, and in mammalian cell culture. Some evidence indicates that EDB can cause sister chromatid exchange and chromosomal aberrations. EDB is a reproductive toxin, but it does not appear to be teratogenic. It has been shown to affect spermatogenesis in rats, bulls and rams and to affect fertility in fowl. Human studies indicate that EDB exposure may harm sperm and decrease fertility. The toxic effect of greatest concern that may result from EDB exposure is cancer. In rats and mice, EDB produced tumors at the application site and at distant sites. When given orally, EDB has produced tumors in the forestomach, lung, and the circulatory system. When administered by inhalation, EDB produced tumors in the nasal cavity, lung, and the circulatory system. Dermal application of EDB produced skin and lung tumors. Analyses of risks from EDB exposure have focused on potential carcinogenic effects. Initial risk estimates, based on animal studies, indicated that citrus workers had essentially a 100% chance of contracting cancer.(ABSTRACT TRUNCATED AT 400 WORDS)

Ethylene Dibromide↗

Analysis of a lysozyme gene from the malaria vector mosquito, Anopheles gambiae.

A genomic DNA sequence encoding a basic lysozyme was isolated from the malaria vector mosquito Anopheles gambiae by screening a library with a probe prepared by PCR of reverse transcribed adult RNA. The sequence consists of an upstream region of about 2 kb, a coding region containing three exons and two introns, and a short 3' untranslated region. The coding region indicates that this mosquito lysozyme consists of a signal peptide of 20 residues followed by an 120 aa mature protein which is very similar to other basic lysozymes. The two small introns, 67 and 76 bp, are located at evolutionarily conserved sites. RT-PCR indicated that this gene is expressed abundantly in sugar-fed adults, and at considerably lower levels when females have fed on blood. Although it remains to be seen whether this gene is induced by bacterial infection, the surrounding sequence contains six sequence motifs very similar to the consensus binding sites for a transcription factor similar to NF-kappa B that are found associated with most insect immune response genes. This lysozyme gene maps to division 27 on the left arm of polytene chromosome 2L. An ORF unrelated to any animal protein in current data bases was found at the 5' end of the clone.

Amino Acid Sequence↗

Genome sequence of the endocellular obligate symbiont of tsetse flies, Wigglesworthia glossinidia.

Many insects that rely on a single food source throughout their developmental cycle harbor beneficial microbes that provide nutrients absent from their restricted diet. Tsetse flies, the vectors of African trypanosomes, feed exclusively on blood and rely on one such intracellular microbe for nutritional provisioning and fecundity. As a result of co-evolution with hosts over millions of years, these mutualists have lost the ability to survive outside the sheltered environment of their host insect cells. We present the complete annotated genome of Wigglesworthia glossinidia brevipalpis, which is composed of one chromosome of 697,724 base pairs (bp) and one small plasmid, called pWig1, of 5,200 bp. Genes involved in the biosynthesis of vitamin metabolites, apparently essential for host nutrition and fecundity, have been retained. Unexpectedly, this obligate's genome bears hallmarks of both parasitic and free-living microbes, and the gene encoding the important regulatory protein DnaA is absent.

Animals↗

A screen for genes that function downstream of Ras1 during Drosophila eye development.

Cell-fate specification of the R7 photoreceptor cell is controlled by the sevenless receptor tyrosine kinase (SevRTK) and Ras1, the Drosophila homologue of mammalian H-ras, K-ras and N-ras oncogenes. An activated form of Ras1 expressed under control of the sevenless enhancer/promoter (sev-Ras1V12) induces production of supernumerary R7 photoreceptor cells, which causes the eye to become rough in appearance. To isolate mutations in genes functioning downstream of Ras1, we carried out a screen for dominant suppressors and enhancers of this rough eye phenotype. Approximately 850,000 mutagenized flies were screened, and 282 dominant suppressors and 577 dominant enhancers were isolated. Mutations in the Drosophila homologues of Raf, MEK, MAPK, type I Geranylgeranyl Transferase and Protein Phosphatase 2A were isolated, as were mutations in several novel signaling genes. Some of these mutant genes appear to be general signaling factors that function in other Ras1 pathways, while one seems to be more specific for photoreceptor development. At least two suppressors appear to function either between Ras1 and Raf or in parallel to Raf.

Animals↗

[Progress in the study of the structure and function of Cre recombinase].

The Cre recombinase, an integrase from bacteriophage P1, catalyzes site-specific recombination between 34-bp repeats termed loxP sites, in the absence of any additional cofactors and energy. Mediated by Cre recombinase, specific DNA fragments can be excised, inversed or integrated depending on the orientation or position of loxP sites in vitro or in vivo. Because of its simplicity and high efficiency, Cre/loxP site-specific recombination system has been widely used in gene deletion and function identification, gene site-specific integration, gene trapping and chromosome engineering. It has been used as a useful tool for DNA recombination in transgenic yeast, plants, insects and mammals. Here progress in the study of the structure and function of Cre recombinase is discussed.

Integrases↗

Sequence analysis of a 33.1 kb fragment from the left arm of Saccharomyces cerevisiae chromosome X, including putative proteins with leucine zippers, a fungal Zn(II)2-Cys6 binuclear cluster domain and a putative alpha 2-SCB-alpha 2 binding site.

In the framework of the European BIOTECH project for sequencing the Saccharomyces cerevisiae genome, we have determined the nucleotide sequence of the left part of the cosmid clone 232 and the cosmid clone 233 provided by F. Galibert (Rennes Cedex, France). We present here 33,099 base pairs of sequence derived from the left arm of chromosome X of strain S288C. This sequence reveals 17 open reading frames (ORFs) with more than 299 base pairs, including the published sequences for ARG3, LIGTR/LIG1, ORF2, ACT3 and SCP160. Two other ORFs showed similarity with S. cerevisiae genes: one with the CAN1 gene coding for an arginine permease, and one with genes encoding the family of transcriptional activators containing a fungal Zn(II)2-Cys6 binuclear cluster domain like that found in Ppr1p or Ga14p. Both putative proteins contain a leucine zipper motif, the Can1p homologue has 12 putative membrane-spanning domains and a putative alpha 2-SCB-alpha 2 binding site. In a diploid disruption mutant of ORF J0922 coding for the transcriptional activator homologue, no colonies appeared before 10 days after transformation and then grew slowly. In contrast, haploid disruption mutants showed a growth phenotype like wild-type cells. One ORF showed weak similarity to the rad4 gene product of Schizosaccharomyces pombe and is essential for yeast growth. Five ORFs showed similarity to putative genes on the right arm of chromosome XI of S. cerevisiae. Two of them have similarity to each other and belong to a family of extracellular proteins that groups mammalian SCP/Tpx-1, insects Ag3/Ag5, plants PR-1 and fungi Sc7/Sc14.(ABSTRACT TRUNCATED AT 250 WORDS)

Chromosomes, Fungal↗

Monandry (monogamy) in natural populations of anopheline mosquitoes.

Polymorphic Y chromosomes within two species of anopheline mosquitoes provide markers for testing if females are inseminated by one or more males in nature. Wild-caught females of Anopheles dirus (sp. A) and An. maculatus s.s. produced 291 and 55 families, respectively, which showed a single type of Y chromosome. One family of the former species showed two types of Y chromosomes. These field data support the idea, established from laboratory studies, that female mosquitoes are largely monandrous (monogamous). Such information is important in interpretation of population biological data and, practically, in attempts to control insect pests by use of genetically designed males.

Animals↗

The Drosophila sex determination signal: how do flies count to two?

Seventy years after the discovery that sex in Drosophila melanogaster is determined by the balance between X chromosomes and autosomes, we can finally identify some of the specific genes whose relative dosage is responsible for the male/female decision in somatic cells and study how they act at the molecular level. Discovery of these sex determination genes was delayed because their mutant phenotypes were unanticipated. It now seems appropriate to consider how the concept of the X/A balance may have limited thinking about the fruit fly sex determination signal.

Animals↗

Biological functions of the ISWI chromatin remodeling complex NURF.

The nucleosome remodeling factor (NURF) is one of several ISWI-containing protein complexes that catalyze ATP-dependent nucleosome sliding and facilitate transcription of chromatin in vitro. To establish the physiological requirements of NURF, and to distinguish NURF genetically from other ISWI-containing complexes, we isolated mutations in the gene encoding the large NURF subunit, nurf301. We confirm that NURF is required for transcription activation in vivo. In animals lacking NURF301, heat-shock transcription factor binding to and transcription of the hsp70 and hsp26 genes are impaired. Additionally, we show that NURF is required for homeotic gene expression. Consistent with this, nurf301 mutants recapitulate the phenotypes of Enhancer of bithorax, a positive regulator of the Bithorax-Complex previously localized to the same genetic interval. Finally, mutants in NURF subunits exhibit neoplastic transformation of larval blood cells that causes melanotic tumors to form.

Adenosine Triphosphatases↗

Modification of the Drosophila heterochromatic mutation brownDominant by linkage alterations.

The variegating mutation brownDominant (bwD) of Drosophila melanogaster is associated with an insertion of heterochromatin into chromosome arm 2R at 59E, the site of the bw gene. Mutagenesis produced 150 dominant suppressors of bwD variegation. These fall into two classes: unlinked suppressors, which also suppress other variegating mutations; and linked chromosome rearrangements, which suppress only bwD. Some rearrangements are broken at 59E, and so might directly interfere with variegation caused by the heterochromatic insertion at that site. However, most rearrangements are translocations broken proximal to bw within the 52D-57D region of 2R. Translocation breakpoints on the X chromosome are scattered throughout the X euchromatin, while those on chromosome 3 are confined to the tips. This suggests that a special property of the X chromosome suppresses bwD variegation, as does a distal autosomal location. Conversely, two enhancers of bwD are caused by translocations from the same part of 2R to proximal heterochromatin, bringing the bwD heterochromatic insertion close to the chromocenter with which it strongly associates. These results support the notion that heterochromatin formation at a genetic locus depends on its location within the nucleus.

Animals↗

Functional expression of insecticide-resistant GABA receptors from the mosquito Aedes aegypti.

We are interested in cloning insecticide resistance genes from vector mosquitos for use as selectable markers in their genetic transformation. As a first step towards this goal, we here report the functional homomultimeric expression of a gamma-aminobutyric acid (GABA) receptor subunit gene, Resistance to dieldrin (Rdl), from the yellow fever mosquito Aedes aegypti in baculovirus-infected insect cell lines. Replacement of alanine296 with a serine leads to approximately 100-fold insensitivity to picrotoxin as previously observed in Drosophila. This shows not only that the mosquito GABA receptor cDNA is functional but also that it can be simply mutated to resistance. Strategies for incorporation of this cDNA into a minigene for the genetic transformation of mosquitoes are discussed.

Aedes↗

Polydnaviruses: potent mediators of host insect immune dysfunction.

Endoparasitic insects are used as biological control agents to kill many species of insect pest. One key to the success of parasitoids that develop in the hemocoel of their host is their ability to knock out the host's immune system, inducing a decline in the responsiveness of a variety of cellular and humoral components so that parasitoid eggs are not encapsulated. In many species parasitized by braconid and ichneumonid wasps, host immunosuppression appears to be mediated by polydnaviruses (PDVs) injected by the female parasitoid into the host hemocoel. The viruses exhibit a complex and intimate genetic relationship with the wasp, since viral sequences are integrated within the wasp's chromosomal DNA. Here Mark Lavine and Nancy Beckage summarize the current evidence for mechanisms of virally induced host immunosuppression in parasitized insects, as well as the roles of other factors including wasp ovarian proteins and venom components, in suppressing hemocyte-mediated and humoral immune responses. Interestingly, in some species, the PDV-induced host immunosuppression appears transitory, with older parasitoid larvae probably exploiting other mechanisms to protect themselves from the host's immune system during the final stages of parasitism. During the final stages of parasitism, the parasitoids likely exploit other mechanisms of immunoevasion via antigen masking, antigen mimicry, or production of active inhibitors of the hemocyte-mediated encapsulation response as well as inhibiting melanization.

Journal Article↗

[Effect of induced chromosomal rearrangements on the fertility and viability of Anopheles atroparvus].

Partial sterility in 3 strains of malarial mosquito Anopheles atroparvus with heterozygous chromosomal rearrangements In2(R;L) 1+In2L1; In3(R;L) 1+D3R1; T(2R;3R)1 was studied. Fertility of these lines was decreased 2.5 to 5 times, as compared with the control. No homozygotes for rearrangements were detected. Causes for partial sterility of insects with heterozygous insertions, translocations and the significance of constructing homozygous strains are discussed.

Animals↗

Quantitative trait loci define genes and pathways underlying genetic variation in longevity.

Quantitative trait locus (QTL) mapping provides a means to discover and roughly position regions of the genome that harbor genes responsible for natural variation in a complex trait. QTL mapping has been utilized extensively in the pursuit of genes contributing to longevity, chiefly in two animal models, the nematode Caenorhabditis elegans and the dipteran insect Drosophila melanogaster. Research on both species has demonstrated that a relatively small set of loci accounts for most of their genetic variance in lifespan. QTL mapping complements the discovery of longevity genes by mutagenesis screens, because the two procedures are predicted to unveil overlapping but distinct types of genes. We argue that information gained from animal models, even invertebrates, can greatly facilitate the process of gene identification and testing of homologous genes in humans.

Aging↗

[Frequency of chromosome aberrations after irradiation of gametes of cabbage root fly (Delia brassicae Bouche) with various doses of X rays].

Chromosome rearrangements were isolated via crossing of F1 offspring from X-irradiated male and female Delia brassicae with intact insects. An enhanced (more than 30%) rate of late embryonic lethality (LEL) in clutches was the primary criterion of rearrangements. When males alone were irradiated at 2.5, 5, and 15 Gy, a total of 7.8, 9.6, and 23.6% of their offspring, respectively, inherited semisterility. After irradiation of females alone, the LEL rate in F1 was insignificant. Hybridization analysis of F2 revealed 18 lines with autosomal inheritance of semisterility. Fourteen lines (43.7%) exhibited an increased rate of LEL linked to the male sex.

Animals↗

Extreme rates and heterogeneity in insect DNA evolution.

DNA-DNA hybridization studies of insects, more specifically Drosophila and cave crickets, have revealed interesting patterns of genome evolution that contrast markedly with what has been seen in other taxa, especially mammals and birds. Insect genomes are composed of sections of single-copy DNA with extreme variation in rates of evolutionary change. This variation is more extreme than between introns and exons; introns fall into the relatively conserved fraction of the genome. Attempts to calculate absolute rates of change in Drosophila DNA have all led to estimates some 5-10 times faster than those found in most vertebrates; this is true even for the more conservative part of the nuclear genome. Finally we point out that morphological similarity, chromosomal similarity, and/or ability to form interspecific hybrids is often associated with quite high levels of single-copy DNA divergence in insects as compared to mammals and birds.

Animals↗