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At least 1,315 records · Page 73Linked to original sources
PubMed1998

The RXR homolog ultraspiracle is an essential component of the Drosophila ecdysone receptor.

Pulses of the steroid hormone ecdysone function as key temporal signals during insect development, coordinating the major postembryonic developmental transitions, including molting and metamorphosis. In vitro studies have demonstrated that the EcR ecdysone receptor requires an RXR heterodimer partner for its activity, encoded by the ultraspiracle (usp) locus. We show here that usp exerts no apparent function in mid-third instar larvae, when a regulatory hierarchy prepares the animal for the onset of metamorphosis. Rather, usp is required in late third instar larvae for appropriate developmental and transcriptional responses to the ecdysone pulse that triggers puparium formation. The imaginal discs in usp mutants begin to evert but do not elongate or differentiate, the larval midgut and salivary glands fail to undergo programmed cell death and the adult midgut fails to form. Consistent with these developmental phenotypes, usp mutants show pleiotropic defects in ecdysone-regulated gene expression at the larval-prepupal transition. usp mutants also recapitulate aspects of a larval molt at puparium formation, forming a supernumerary cuticle. These observations indicate that usp is required for ecdysone receptor activity in vivo, demonstrate that the EcR/USP heterodimer functions in a stage-specific manner during the onset of metamorphosis and implicate a role for usp in the decision to molt or pupariate in response to ecdysone pulses during larval development.

Animals↗
PubMed2005

Characterization of deltamethrin resistance in field populations of Aedes aegypti in Thailand.

Five field collections of adult Aedes aegypti mosquitoes from different areas in Bangkok and Pathum Thani provinces were subjected to susceptibility tests against deltamethrin. Low levels of resistance were detected among all populations tested (RR50 = 8-17.2) compared to the susceptible strain, Bora (French Polynesia). Among the five populations tested, the BKH (Bang Khen, Bangkok) and PSC (Phasicharoen, Bangkok) populations showed a higher level of deltamethrin resistance than the other three populations (RR50 of BKH = 17.2, and of PSC= 13.6) and cross-resistance to DDT was observed in these strains. Biochemical analysis showed a significant elevation of mixed function oxidases enzyme activity in all populations. There was an elevation of non-specific esterases in all populations except BKL, and there was no consistent association of glutathione S-transferases with deltamethrin and DDT resistance, although not all populations were bioassayed for DDT. The partial cDNA sequence of the para-type voltage-dependent sodium channel (IIS4-IIS6) was determined for BKH and PSC populations. Common amino acid substitution, leucine to phenylalanine in the IIS6 region, found for insects including Anopheles gambiae was not found in either the BKH or the PSC populations. However, two other amino acid substitutions (proline substituted with serine at position 64 in the PSC population and leucine with phenylalanine at position 69 in the BKH population) were found in the IIS5-IIS6 inter-segment region sequenced. The role these substitutions play in target site resistance is uncertain at this time.

Aedes↗
PubMed2001

Structure and evolution of the mitochondrial DNA complete control region in the Drosophila subobscura subgroup.

The complete A + T-rich region of mitochondrial DNA (mtDNA) has been cloned and sequenced in the species of the Drosophila subobscura subgroup D. subobscura, D. madeirensis and D. guanche. Comparative analysis of these sequences with others already published has identified new sequence motifs that are conserved in Drosophila and other insects. A putative bi-directional promoter and a stop signal are proposed to be involved in the primary mtDNA strand replication of Drosophila. This region strongly resolves relationships of the species included in a phylogenetic analysis, both for closely related species and also at deeper phylogenetic levels when only the left and central domains are taken into account.

AT Rich Sequence↗
PubMed1995

Epithelial planar polarity in the developing Drosophila eye.

Experiments with the insect ectoderm have suggested that planar polarity in epithelia results from the local orientation of cells to the slope of a gradient of positional information. Here we show that planar polarity in the Drosophila eye is inverted when the morphogenetic wave that sweeps through the presumptive retinal epithelium is induced to move in the reverse direction. We suggest that the movement of the morphogenetic wave may be causal in establishing the planar polarity of this epithelium.

Animals↗
PubMed2001

Homeotic evidence for the appendicular origin of the labrum in Tribolium castaneum.

The ontogeny of the insect labrum, or upper lip, has been debated for nearly a century. Recent molecular data suggest a segmental appendage origin of this structure. Here we report the first arthropod mutation associated with a homeotic transformation of the labrum. Antennagalea-5 (Ag(5)) transforms both antennal and labral structures to resemble those of gnathal appendages in Tribolium castaneum. This labral transformation suggests that the labrum is a fused structure composed of two pairs of appendage endites, and is serially homologous to the gnathal appendages.

Animals↗
PubMedDate not supplied

Molecular cloning of a female-specific cDNA with unique repeat sequences from the fat body of the adult locust, Locusta migratoria.

A cDNA clone encoding a 25-kDa protein (25K) was isolated from a cDNA library made from RNA isolated from the adult fat body and ovaries of the locust, Locusta migratoria. The longest open reading frame of this cDNA clone encodes a 225-amino acid polypeptide, the N-terminal end of which was similar to the 21-kDa and 19-kDa juvenile hormone induced proteins identified in the locust hemolymph, but the C-terminal end was different. The C-terminal end of the 25K cDNA contained seven unique repeat elements of 10 amino acids each, most of which are polar residues. Expression of the 25K mRNA was tissue-, development- and sex-specific. A 1.2-kb mRNA was detected using the 25K cDNA as a probe only in the fat body of adult females. The mRNA started to appear at day 4 after the insect molted to the adult and rapidly increased by day 6. The mRNA was absent in the ovarian follicle cells and fat body of adult males. In vitro transcription and translation of the 25K cDNA produced a protein that migrated around 32 kDa on sodium dodecyl sulfate polyacrylamide gels. The 25K cDNA was expressed in a baculovirus expression system and the protein produced also migrated around 32 kDa.

Amino Acid Sequence↗
PubMed2001

Chromosome races with Pliocene origins: evidence from mtDNA.

There are eight distinct chromosomal races of the New Zealand weta Hemideina thoracica. We used mtDNA sequence data to test the hypothesis that these races originated on islands during the early Pliocene (7--4 million years ago). Nine major mitochondrial lineages were identified from 65 cytochrome oxidase I sequences. Phylogenetic analysis of these lineages suggests that they arose at approximately the same time. The geographical distribution of some lineages coincides with areas that were islands during the Pliocene. Overall, hierarchical AMOVA analysis shows that chromosomal races and Pliocene islands describe only 28% and 24%, respectively, of the total current mtDNA variation. However, removing one widespread (A) and one putatively introgressed (F) lineage increases these estimates to 65% and 80%, respectively. Intraspecific sequence divergence was very high, reaching a maximum of 9.5% (uncorrected distance) and GC content was high compared to other insect mtDNA sequences. Average corrected distance among mtDNA lineages supports the Pliocene origins of this level of genetic diversity. In the southern part of the species range there is reduced mtDNA variation, probably related to local extinction of H. thoracica populations from recent volcanic activity and subsequent re-colonization from a leading edge. In contrast, in this southern part there are five chromosome races, suggesting that chromosome races here may be younger than those in the north.

Animals↗
PubMed2003

Juvenile hormone acid methyltransferase: a key regulatory enzyme for insect metamorphosis.

Juvenile hormone (JH) acid methyltransferase (JHAMT) is an enzyme that converts JH acids or inactive precursors of JHs to active JHs at the final step of JH biosynthesis pathway in insects. By fluorescent mRNA differential display, we have cloned a cDNA encoding JHAMT from the corpora allata (CA) of the silkworm, Bombyx mori (BmJHAMT). The BmJHAMT cDNA encodes an ORF of 278 aa with a calculated molecular mass of 32,544 Da. The predicted amino acid sequence contains a conserved S-adenosyl-l-methionine (SAM) binding motif found in the family of SAM-dependent methyltransferases. Purified N-terminal 6xHis-tagged recombinant BmJHAMT protein expressed in Escherichia coli catalyzed conversion of farnesoic acid and JH acids I, II, and III to their cognate methyl esters in the presence of SAM, confirming that this cDNA encodes a functional JHAMT. Putative orthologs, DmJHAMT and AgJHAMT, were identified from the genome sequence of the fruit fly Drosophila melanogaster, and a malaria vector, Anopheles gambiae, respectively. Northern blot and quantitative RT-PCR analyses revealed that the BmJHAMT gene was expressed specifically in the CA throughout the third and fourth instar. At the beginning of the last (fifth) instar, the expression level of BmJHAMT declined rapidly and became undetectable by day 4 and remained so until pupation. Correlation of the BmJHAMT gene expression and the JH biosynthetic activity in the CA suggests that the transcriptional suppression of the BmJHAMT gene is crucial for the termination of JH biosynthesis in the CA, which is a prerequisite for the initiation of metamorphosis.

Amino Acid Sequence↗
PubMed2002

Identification in Drosophila melanogaster of the invertebrate G protein-coupled FMRFamide receptor.

We here describe the cloning and characterization of the functionally active Drosophila melanogaster (Drm) FMRFamide receptor, which we designated as DrmFMRFa-R. The full-length ORF of a D. melanogaster orphan receptor, CG 2114 (Berkeley Drosophila Genome Project), was cloned from genomic DNA. This receptor is distantly related to mammalian thyroid-stimulating hormone-releasing hormone receptors and to a set of Caenorhabditis elegans orphan receptors. An extract of 5,000 central nervous systems from the related but bigger flesh fly, Neobellieria bullata (Neb), was used to screen cells expressing the orphan receptor. Successive purification steps, followed by MS, revealed the sequence of two previously uncharacterized endogenous peptides, APPQPSDNFIRFamide (Neb-FIRFamide) and pQPSQDFMRFamide (Neb-FMRFamide). These are reminiscent of other insect FMRFamide peptides, having neurohormonal as well as neurotransmitter functions. Nanomolar concentrations of the Drm FMRFamides (DPKQDFMRFamide, TPAEDFMRFamide, SDNFMRFamide, SPKQDFMRFamide, and PDNFMRFamide) activated the cognate receptor in a dose-dependent manner. To our knowledge, the cloned DrmFMRFa-R is the first functionally active FMRFamide G protein-coupled receptor described in invertebrates to date.

Amino Acid Sequence↗
PubMed1997

Rapid cloning of insect transposon insertion junctions using 'universal' PCR.

Very highly degenerate primers with short specific 3' anchor sequences and 5' adaptors were used in conjunction with nested specific primers to amplify large numbers of unknown insertion junctions of the insect retrotransposon Woot, using genomic DNA as template for the polymerase chain reaction (PCR). This technique, sometimes referred to as universal PCR, is a powerful method for molecular characterization of transposon insertions into genomes, and more generally for short-distance chromosome walking through unknown DNA. Twenty-four unique insertion junctions were cloned and sequenced from two strains of Tribolium castaneum and one strain of T. freemani. Inspection of these sequences revealed that integration of the Woot retrotransposon is cued by the insertion target motif, GTAC, in both species.

Animals↗
PubMed1994

Complete base sequence for the mitochondrial large subunit ribosomal RNA of the gypsy moth Lymantria dispar (L.).

A 1355 bp sequence (accession number L32141) isolated from a gypsy moth (Lymantria dispar) cDNA library showed 68-74% sequence identity to mitochondrial large subunit ribosomal RNA (mt IrRNA) sequences of Locusta migratoria, Apis mellifera, Aedes albopictus, Anopheles gambiae and two Drosophila species. A comparison of the primary sequences of the mt IrRNAs from the above insects in four orders and from Esherichia coli demonstrated regions of conservation which presumably correspond to regions of functional and/or structural homology. A secondary structure for the gypsy moth mt IrRNA sequence was derived based on the proposed secondary structures of Drosophila yakuba and Aedes albopictus mt IrRNAs (Gutell & Fox, Nucleic Acid Res 16 (Suppl.), r175-r269, 1988). This sequence was found to hybridize to about 10-15% of the clones in several (eleven) gypsy moth cDNA libraries.

Animals↗
PubMed2001

Genomic gigantism: DNA loss is slow in mountain grasshoppers.

Several studies have shown DNA loss to be inversely correlated with genome size in animals. These studies include a comparison between Drosophila and the cricket, Laupala, but there has been no assessment of DNA loss in insects with very large genomes. Podisma pedestris, the brown mountain grasshopper, has a genome over 100 times as large as that of Drosophila and 10 times as large as that of Laupala. We used 58 paralogous nuclear pseudogenes of mitochondrial origin to study the characteristics of insertion, deletion, and point substitution in P. pedestris and Italopodisma. In animals, these pseudogenes are "dead on arrival"; they are abundant in many different eukaryotes, and their mitochondrial origin simplifies the identification of point substitutions accumulated in nuclear pseudogene lineages. There appears to be a mononucleotide repeat within the 643-bp pseudogene sequence studied that acts as a strong hot spot for insertions or deletions (indels). Because the data for other insect species did not contain such an unusual region, hot spots were excluded from species comparisons. The rate of DNA loss relative to point substitution appears to be considerably and significantly lower in the grasshoppers studied than in Drosophila or Laupala. This suggests that the inverse correlation between genome size and the rate of DNA loss can be extended to comparisons between insects with large or gigantic genomes (i.e., Laupala and Podisma). The low rate of DNA loss implies that in grasshoppers, the accumulation of point mutations is a more potent force for obscuring ancient pseudogenes than their loss through indel accumulation, whereas the reverse is true for Drosophila. The main factor contributing to the difference in the rates of DNA loss estimated for grasshoppers, crickets, and Drosophila appears to be deletion size. Large deletions are relatively rare in Podisma and Italopodisma.

Animals↗
PubMed2004

Compensated deleterious mutations in insect genomes.

Relatively little is known about the importance of amino acid interactions in protein and phenotypic evolution. Here we examine whether mutations that are pathogenic in Drosophila melanogaster become fixed via epistasis in other Dipteran genomes. Overall divergence at pathogenic amino acid sites is reduced. However, approximately 10% of the substitutions at these sites carry the exact same pathogenic amino acid found in D. melanogaster mutants. Hence compensatory mutation(s) must have evolved. Surprisingly, the fraction 10% is not affected by phylogenetic distance. These results support a selection-driven process that allows compensated amino acid substitutions to become rapidly fixed in taxa with large populations.

Amino Acid Sequence↗
PubMed2000

Molecular characterization of a prophenoloxidase cDNA from the malaria mosquito Anopheles stephensi.

Some refractory anopheline mosquitoes are capable of killing Plasmodium, the causative agent of malaria, by melanotic encapsulation of invading ookinetes. Phenoloxidase (PO) appears to be involved in the formation of melanin and toxic metabolites in the surrounding capsule. A cDNA encoding Anopheles stephensi prophenoloxidase (Ans-proPO) was isolated from a cDNA library screened with an amplimer produced by reverse transcriptase polymerase chain reaction (RT-PCR) with degenerate primers designed against conserved proPO sequences. The 2.4-kb-long cDNA has a 2058 bp open reading frame encoding Ans-proPO of 686 amino acids. The deduced amino acid sequence shows significant homology to other insect proPO sequences especially at the two putative copper-binding domains. In A. stephensi, Ans-proPO expression was detected in larval, pupal and adult stages. The Ans-proPO mRNA was detected by RT-PCR and in situ hybridization in haemocytes, fat body and epidermis of adult female mosquitoes. A low level of expression was detected in the ovaries, whereas no expression was detected in the midguts. Semi-quantitative RT-PCR analysis of Ans-proPO mRNA showed that its expression was similar in adult female heads, thoraxes and abdomens. No change in the level of Ans-proPO expression was found in adult females after blood feeding, bacterial challenge or Plasmodium berghei infection. However, elevated PO activity was detected in P. berghei-infected mosquitoes, suggesting that in non-selected permissive mosquitoes PO may be involved in limiting parasite infection. Genomic Southern blot and immunoblots suggest the presence of more than one proPO gene in the A. stephensi genome, which is consistent with the findings in other Diptera and Lepidoptera species. The greatest similarity in sequence and expression profile between Ans-proPO and A. gambiae proPO6 suggests that they might be homologues. Our results demonstrate that Ans-proPO is constitutively expressed through different developmental stages and under different physiological conditions, implying that other factors in the proPO activation cascade regulate melanotic encapsulation.

Amino Acid Sequence↗
PubMed1997

Evolution of R1 and R2 in the rDNA units of the genus Drosophila.

R1 and R2 are non-long terminal repeat (non-LTR) retrotransposable elements that specifically insert in the 28S ribosomal RNA (rRNA) genes of insects. Using the Drosophila genus, which includes some of the best characterized insect taxa, we have conducted a number of studies on the evolution of these elements. We find that R1 and R2 are subject to the same recombinational forces that give rise to the concerted evolution of the rDNA units. The turnover of R1 and R2 elements can be readily documented in different strains of D. melanogaster using 5' truncated elements as restriction-length polymorphisms. This turnover leads to uniform populations of elements with nucleotide sequence divergence of different copies averaging only 0.23% for the R2 and 0.47% for the R1 elements. Molecular phylogenetic analysis of elements from 16 different species of Drosophila suggests that these elements have been stable components of the rDNA locus for the 50-70 million year history of the Drosophila genus. Using changes at synonymous positions within the protein-encoding regions as estimates of the baseline substitution rate, it could be shown that R1 and R2 are evolving at rates similar to that of typical protein encoding genes provided corrections are made for the low codon bias of the elements. R1 and R2 are clearly well-adapted for their existence in the rDNA units of their host.

Animals↗
PubMed1989

Morphogen gradients and the control of body pattern in insect embryos.

The basic body plan of insects is set up in response to determinants initially localized at the anterior and posterior poles of the egg. Early in development, these determinants give rise to a series of morphogen gradients which in turn trigger a cascade of molecular signals determining the body pattern. This signalling process is outlined and recent experiments testing the roles of these gradient systems in determining anterior and posterior pattern are described. The results of these experiments suggest that anterior pattern is controlled by a single instructive gradient, whereas posterior pattern depends on the overlap of several gradients, each providing only one or a few distinct responses.

Animals↗
PubMed2003

Recent horizontal transfer of mellifera subfamily mariner transposons into insect lineages representing four different orders shows that selection acts only during horizontal transfer.

We report the isolation and sequencing of genomic copies of mariner transposons involved in recent horizontal transfers into the genomes of the European earwig, Forficula auricularia; the European honey bee, Apis mellifera; the Mediterranean fruit fly, Ceratitis capitata; and a blister beetle, Epicauta funebris, insects from four different orders. These elements are in the mellifera subfamily and are the second documented example of full-length mariner elements involved in this kind of phenomenon. We applied maximum likelihood methods to the coding sequences and determined that the copies in each genome were evolving neutrally, whereas reconstructed ancestral coding sequences appeared to be under selection, which strengthens our previous hypothesis that the primary selective constraint on mariner sequence evolution is the act of horizontal transfer between genomes.

Amino Acid Sequence↗
PubMed1995

Molecular cloning and characterization of cDNA for insect biogenic peptide, growth-blocking peptide.

Growth-blocking peptide (GBP) is an insect biogenic peptide that prevents the onset of metamorphosis from larva to pupa. A cDNA coding for GBP is described. Mixed oligonucleotides derived from a GBP peptide sequence were used to generate amplified DNA by the polymerase chain reaction (PCR). Based on the sequence of the amplified DNA, a 41 bases oligonucleotide was designed for screening a cDNA library which was constructed from the armyworm Pseudaletia separata larvae parasitized with the parasitic wasp Cotesia kariyai. The cloned cDNA for GBP was 809 base pairs in length. An open reading frame of 429 base pairs encodes a pre-pro-peptide of 143 amino acid residues in which GBP is localized at the C-terminal region, and other three peptides including a putative signal peptide and appropriate processing sites for endoproteolytic cleavage precede the GBP sequence. Northern blot analyses demonstrate the presence of a 800-base mRNA transcript in fat body and 2.5-kilobase transcript in brain and nerve cord, suggesting the possibility that the transcription of GBP gene is regulated in a tissue-dependent manner. This interpretation was supported by isolating a GBP cDNA fragment from cDNA pool of brain-nerve cords. GBP mRNA is constantly expressed in both parasitized and non-parasitized last instar larvae and there is no difference in the levels of the mRNA between both larvae, thus indicating that parasitism may effect on translational or posttranslational level to elevate plasma GBP concentration.

Amino Acid Sequence↗