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P element homing to the Drosophila bithorax complex.

P elements containing a 7 kb DNA fragment from the middle of the Drosophila bithorax complex insert preferentially into the bithorax complex or into the adjacent chromosome regions. This 'homing' property is similar to that reported for the engrailed promoter (Hama, C., Ali, Z. and Kornberg, T. B. (1990) Genes Dev. 4, 1079-1093). The 7 kb fragment does not contain any known promoter, but it acts as a boundary element separating adjacent segmental domains. An enhancer-trap P element was constructed with the homing fragment and the selectable marker flanked by FRT sites. P insertions can be trimmed down by Flp-mediated recombination to just the lacZ reporter, so that the (beta)-galactosidase pattern is not influenced by sequences inside the P element. Twenty insertions into the bithorax complex express (beta)-galactosidase in segmentally limited patterns, reflecting the segmental domains of the bithorax complex where the elements reside. The mapping of segmental domains has now been revised, with enlargement of the abx/bx, bxd/pbx, and the iab-3 domains. The FRT sites in the P elements permit recombination between pairs of elements on opposite chromosomes, to generate duplications or deletions of the DNA between the two insertion sites. Using this technique, the length of the Ultrabithorax transcription unit was varied from 37 to 138 kb, but there was surprisingly little effect on Ultrabithorax function.

Animals↗

Detection and molecular analysis of plant- and insect-associated bacteria harboring aconitate isomerase involved in biosynthesis of trans-aconitic acid as antifeedant in brown planthoppers.

The activity of aconitate isomerase, which is involved in the biosynthesis of trans-aconitic acid as antifeedant in brown planthoppers, was detected in Pseudomonas fluorescens LRB3W1 and Pseudomonas putida MAFF301685 but not in Pseudomonas putida MAFF301684. The enzyme activity was induced in the presence of trans-aconitate, and therefore bacteria showing the enzyme activity were easily detected by their ability to grow on the minimal medium containing trans-aconitate as the sole carbon source (ACO agar medium). Experiments on growth of plant- or insect-associated bacteria on ACO agar medium showed that most of the Gram-negative bacteria displayed the aconitate isomerase activity unlike most of the Gram-positive bacteria isolated mainly from insects. Mini-Tn5 transposon derivatives of P. fluorescens LRB3W1 lacking completely or partially their ability to grow on ACO agar medium were obtained. Southern blot analysis with a mini-Tn5 DNA probe definitely showed that the genes responsible for the biosynthesis of aconitate isomerase present on chromosomal DNA. Thus, it was suggested that genes for aconitate isomerase biosynthesis are commonly present in Gram-negative plant- or insect-associated bacteria, and also the DNA fragments including the genes were detected in P. fluorescens LRB3W1.

Aconitic Acid↗

ERGA-BGE chromosome-level genome assembly of the giant stream lacewing  Osmylus fulvicephalus (Scopoli, 1763).

The giant stream lacewing, Osmylus fulvicephalus (Scopoli, 1763), is a widespread European species belonging to the insect order Neuroptera. Its cryptic larvae are predators found at the banks of streams and smaller rivers where they use their piercing, lance-shaped stylets to inject venom into their arthropod prey. Here, we present the reference genome of the giant stream lacewing as a crucial resource for uncovering the genetic basis of venom evolution in Neuroptera. The chromosome-level genome encompasses 674.7 Mb and is composed of 60 contigs and 24 scaffolds where 99.2% of the assembly is distributed among the 6 contiguous chromosomal pseudomolecules and two sex chromosomes (X and Y). Contig and scaffold N50 have a value of 51.5 Mb and 116.2 Mb, respectively. This reference genome is the first genomic resource from the family of lance lacewings, providing valuable data for clarifying the phylogenetic placement of the family Osmylidae within Neuroptera.

Biodiversity Genomics Europe↗

[Heterochromatic ABO locus of the Drosophila melanogaster X chromosome overlaps with the region of localization of repeats, including mobile elements and Stellate genes].

A loss of certain heterochromatic regions (ABO loci) of various chromosomes dramatically distorts the early embryo development in the progeny of females mutant for the abnormal oocyte (abo) gene, which is located in euchromatin of chromosome 2. One ABO locus (X-ABO) is in X-chromosomal heterochromatin distal of the nucleolus organizer. A cluster of the Stellate repeats is located in the same heterochromatin block. Deletions of various fragments from distal heterochromatin were tested for the effect on expression of the abo mutation. The X-ABO locus was assigned to X-chromosomal heterochromatin segment h26 and shown to include repeats consisting mostly of mobile elements and defective Stellate copies. A major part of the regular Stellate tandem repeats proved to be distal of the X-ABO locus.

ABO Blood-Group System↗

Patterns of Hermes transposition in Drosophila melanogaster.

Transposable elements are being developed as tools for genomics and for the manipulation of insect genotypes for the purposes of biological control. An understanding of their transposition behavior will facilitate the use of these elements. The behavior of an autonomous Hermes transposable element from Musca domestica in the soma and germ-line of Drosophila melanogaster was investigated using the method of transposon display. In the germ-line, Hermes transposed at a rate of approximately 0.03 jumps per element per generation. Within the soma Hermes exhibited markedly non-random patterns of integration. Certain regions of the genome were distinctly preferred over others as integration targets, while other regions were underrepresented among the integration sites used. One particular site accounted for 4.4% of the transpositions recovered in this experiment, all of which were located within a 2.5-kb region of the actin5C promoter. This region was also present within the Hermes element itself, suggesting that this clustering is an example of transposable element "homing". Clusters of integration sites were also observed near the original donor sites; these represent examples of local hopping. The information content (sequence specificity) of the 8-bp target site was low, and the consensus target site resembles that determined from plasmid-based integration assays.

Animals↗

Mutability studies in two Drosophila melanogaster isogenic stocks, endemic for C picornavirus and virus-free.

A virus-free Drosophila stock was obtained by outer disinfection of eggs from DCV-contaminated females. The healthy flies exhibited 3 times less lethal mutations on the X chromosome than did the diseased flies. In addition, on X and 2nd chromosomes, the mutability of the infected males was 2-3 times lower than that of the infected females. The natural viruses of Drosophila are partially responsible for the rate of mutations occurring in the wild populations of this insect.

Animals↗

Dosage compensation of the Drosophila white gene requires both the X chromosome environment and multiple intragenic elements.

The X-linked white gene when transposed to autosomes retains only partial dosage compensation. One copy of the gene in males expresses more than one copy but less than two copies in females. When inserted in ectopic X chromosome sites, the mini-white gene of the CaspeR vector can be fully dosage compensated and can even achieve hyperdosage compensation, meaning that one copy in males gives more expression than two copies in females. As sequences are removed gradually from the 5' end of the gene, we observe a progressive transition from hyperdosage compensation to full dosage compensation to partial dosage compensation. When the deletion reaches -17, the gene can no longer dosage compensate fully even on the X chromosome. A deletion reaching +173, 4 bp preceding the AUG initiation codon, further reduces dosage compensation both on the X chromosome and on autosomes. This truncated gene can still partially dosage compensate on autosomes, indicating the presence of dosage compensation determinants in the protein coding region. We conclude that full dosage compensation requires an X chromosome environment and that the white gene contains multiple dosage-compensation determinants, some near the promoter and some in the coding region.

ATP-Binding Cassette Transporters↗

Reduced X-linked nucleotide polymorphism in Drosophila simulans.

Population genetic theory predicts that selectively driven changes of allele frequency for both beneficial and deleterious mutants reduce polymorphism at tightly linked sites. All else being equal, these reductions in polymorphism are expected to be greater when recombination rates are lower. Therefore, the empirical observation of a positive correlation between recombination rates and amounts of DNA polymorphism across the Drosophila melanogaster genome can be explained by two very different types of natural selection. Here, we evaluate alternative models of effects of selection on linked sites by comparison of X-linked and autosomal variation. We present polymorphism data from 40 genes distributed across chromosome arms X and 3R of Drosophila simulans, a sibling species of D. melanogaster. We find significantly less silent polymorphism in D. simulans on the X chromosome than on 3R, but no difference between arms for silent divergence between species. This pattern is incompatible with predictions from theoretical studies on the effect of negative selection on linked sites. We propose that some form of positive selection having greater effects on sex chromosomes than on autosomes is the better explanation for the D. simulans data.

Animals↗

Chromosome-level genome assembly of Ampulex clypecomplana Chen & Li (Hymenoptera: Ampulicidae).

Ampulex clypecomplana Chen & Li, 2010 (Hymenoptera: Ampulicidae) is an important predatory insect in Hymenoptera. However, molecular information about this predatory insect is currently limited. In this study, we employed ONT long-read sequencing, MGI-SEQ short-read sequencing, Hi-C sequencing and transcriptomic data to assemble the high-quality genome of A. clypecomplana. The genome assembly length was 338.43 Mb, with a Scaffold N50 length of 19.05 Mb. Our BUSCO analysis further confirmed the gene coverage completeness of the genome assembly to be 99.2%. Phylogenetic analysis indicated that A. clypecomplana appeared approximately 132 million years ago. We annotated 110.75 Mb of repetitive sequences, accounting for 32.72% of the entire genome. In A. clypecomplana, we identified 180 gene expansions and 1029 genes that underwent contraction or loss. The high-quality genome of A. clypecomplana provides a valuable genetic resource for future research in evolution, molecular biology, and applied studies.

Animals↗

The conserved HR domain of the Drosophila suppressor 2 of zeste [Su(z)2] and murine bmi-1 proteins constitutes a locus-specific chromosome binding domain.

The related Drosophila Suppressor 2 of zeste [Su(z)2] and Posterior sex combs (Psc) proteins are both locus-specific chromosome binding proteins. They are found at many of the same polytene chromosome loci as other Polycomb-group proteins. The 1,365 amino acid Su(z)2 protein and the 1,603 amino acid Psc protein share a conserved 200 amino acid domain, the homology region (HR). To identify the protein domain responsible for locus-specific chromosome binding, we made a series of Hsp70:cDNA deletion constructs of the Sz(z)2 gene and transformed these into flies. We found that the HR is necessary and sufficient for Su(z)2 locus-specific polytene chromosome binding. The murine Bmi-1 protein also shares the conserved HR domain. When expressed in flies, the Bmi-1 protein showed a locus-specific chromosome binding pattern similar to that of the Su(z)2 and Psc proteins. These results argue that a locus-specific chromosome binding function resides in the HR domain. Other results show that a second, low affinity, non-specific chromosome binding function is localized outside the HR in the Su(z)2 protein, and that the Su(z)2 protein contains at least two nuclear localization signals.

Animals↗

Molecular characterization of Tclabial and the 3' end of the Tribolium homeotic complex.

The homeotic selector gene labial is located at the 3' end of the Antennapedia complex (ANTC) and is required for proper head development in Drosophila. We have cloned and characterized the Tribolium ortholog of labial, Tclabial (Tclab). Similar to Drosophila labial, Tclab contains a single large intron upstream of the homeobox. In contrast, Tclab lacks an intron within the homeobox. The Tribolium ortholog of chaoptic, Tcchaoptic, transcribed from the opposite strand, is located immediately downstream of the homeotic complex, and its 3'UTR overlaps that of Tclab by 50 nucleotides. We have also sequenced the 13.5 kb interval between Tclab and maxillopedia (the Tribolium ortholog of Drosophila proboscipedia). In contrast to Drosophila, there is not a cluster of cuticle genes in this region. Finally, we have examined the expression of Tclab transcripts in Tribolium embryos. As previously described for Drosophila and other insects, the expression of Tclab is specific to the intercalary segment.

Amino Acid Sequence↗

Polydnavirus-facilitated endoparasite protection against host immune defenses.

The polydnavirus of Campoletis sonorensis has evolved with an unusual life cycle in which the virus exists as an obligate symbiont with the parasite insect and causes significant physiological and developmental alterations in the parasite's host. The segmented polydnavirus genome consists of double-stranded superhelical molecules; each segment is apparently integrated into the chromosomal DNA of each male and female wasp. The virus replicates in the nucleus of calyx cells and is secreted into the oviduct. When the virus is transferred to the host insect during oviposition, gene expression induces host immunosuppression and developmental arrest, which ensures successful development of the immature endoparasite. In the host, polydnavirus expression is detected by 2 hr and during endoparasite development. Most of the abundantly expressed viral genes expressed very early after parasitization belong to multigene families. Among these families, the "cysteine-rich" gene family is the most studied, and it may be important in inducing host manifestations resulting in parasite survival. This gene family is characterized by a similar gene structure with introns at comparable positions within the 5' untranslated sequence and just 5' to a specific cysteine codon (*C) within a cysteine motif, C-*C-CC-C-C. Another unusual feature is that the nucleotide sequences of introns 2 in the subfamily WHv1.0/WHv1.6 are more conserved than those of the flanking exons. The structures of these viral genes and possible functions for their encoded protein are considered within the context of their endoparasite and virus strategy for genetic adaptation and successful parasitization.

Amino Acid Sequence↗

Chitinases are a multi-gene family in Aedes, Anopheles and Drosophila.

Degenerate primers were used to amplify by the polymerase chain reaction (PCR) DNA fragments from the chitinase genes of five insect species: Aedes aegypti, Anopheles freeborni, Anopheles gambiae, Anopheles stephensi and Drosophila melanogaster. As many as four different products were found for each species; each deduced protein sequence having greatest homology to chitinase sequences from other species of insects and the crustacean, Penaeus japonicus. The four PCR products of A. aegypti hybridize to two loci, with three of the products derived from either three tightly linked genes or a single gene with three catalytic domains. Southern blot hybridizations of the PCR products from the species of Anopheles suggest a similar arrangement.

Aedes↗

How flies make one equal two: dosage compensation in Drosophila.

Dosage compensation is the process by which the expression of X-linked genes is equalized in males and females. In Drosophila, dosage compensation occurs by coordinately upregulating the transcription rates of all the genes on the single X chromosome in males. This hypertranscription requires the functioning of four autosomal male-specific lethal (msl) genes and is under the control of the Sxl gene. Recent genetic and molecular studies have suggested that the msl proteins may associate with one another in a sex-specific heteromeric complex on the male X chromosome, where they may function to alter its chromatin structure.

Animals↗

Both naturally occurring insertions of transposable elements and intermediate frequency polymorphisms at the achaete-scute complex are associated with variation in bristle number in Drosophila melanogaster.

A restriction enzyme survey of a 110-kb region including the achaete scute complex (ASC) examined 14 polymorphic molecular markers in a sample of 56 naturally occurring chromosomes. Large insertions as a class were associated with a reduction in both sternopleural and abdominal bristle number, supporting deleterious mutation-selection equilibrium models for the maintenance of quantitative genetic variation. Two polymorphic sites were independently associated with variation in bristle number measured in two genetic backgrounds as assessed by a permutation test. A 6-bp deletion near sc alpha is associated with sternopleural bristle number variation in both sexes and a 3.4-kb insertion between sc beta and sc gamma is associated with abdominal bristle number variation in females. Under an additive genetic model, the small deletion polymorphism near sc alpha accounts for 25% of the total X chromosome genetic variation in sternopleural bristle number, and the 3.4 kb insertion accounts for 22% of the total X chromosome variation in female abdominal bristle number. The observation of common polymorphisms associated with variation in bristle number is more parsimoniously explained by models that incorporate balancing selection or assume variants affecting bristle number are neutral, than mutation-selection equilibrium models.

Animals↗

Ordered assembly of roX RNAs into MSL complexes on the dosage-compensated X chromosome in Drosophila.

BACKGROUND: In the male Drosophila, the X chromosome is transcriptionally upregulated to achieve dosage compensation, in a process that depends on association of the MSL proteins with the X chromosome. A role for non-coding RNAs has been suggested in recent studies. The roX1 and roX2 RNAs are male-specific, non-coding RNAs that are produced by, and also found associated with, the dosage-compensated male X chromosome. Whether roX RNAs are physically part of the MSL complex has not been resolved. RESULTS: We found that roX RNAs colocalize with the MSL proteins and are highly unstable unless the MSL complex is coexpressed, suggesting a physical interaction. We were able to immunoprecipitate roX2 RNA from male tissue-culture cells with antibodies to the proteins Msl1 and Mle, consistent with an integral association with MSL complexes. Localization of roX1 and roX2 RNAs in mutants indicated an order of MSL-complex assembly in which roX2 RNA is incorporated early in a process requiring the Mle helicase. We also found that the roX2 gene, like roX1, is a nucleation site for MSL complex spreading into flanking chromatin in cis. CONCLUSIONS: Our results support a model in which MSL proteins assemble at specific chromatin entry sites (including the roX1 and roX2 genes); the roX RNAs join the complex at their sites of synthesis; and complete complexes spread in cis to dosage compensate most genes on the X chromosome.

Animals↗