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Evidence of hormonal control of the nucleolar activity in Sciara ocellaris.

The nucleolus organizing region in the salivary gland of Sciara ocellaris becomes gradually less active as the larvae approach pupation. The cytological visible condensation of the nucleolar puff is paralleled by a decrease in the incorporation of [3H]-uridine into the ribosomal RNA. Alterations in the nucleolar behavior were brought about by extirpation of the neuroendocrine glands at six stages of development. There is a 'critical stage' during the 7th day of the 4th instar when the hormonal stimulus for pupation occurs. If the ligature is performed before this stage the NOR remains expanded and the larval-pupal ecdysis is inhibited; if done later on the NOR condenses at a normal rate and the ligated larvae mount to head-less pupae. These results suggest that inhibition of rRNA synthesis and consequent condensation of the NOR is under the control of the moulting hormone.

Animals↗

Drosophila homologs of baculovirus inhibitor of apoptosis proteins function to block cell death.

Apoptotic cell death is a mechanism by which organisms eliminate superfluous or harmful cells. Expression of the cell death regulatory protein REAPER (RPR) in the developing Drosophila eye results in a small eye owing to excess cell death. We show that mutations in thread (th) are dominant enhancers of RPR-induced cell death and that th encodes a protein homologous to baculovirus inhibitors of apoptosis (IAPs), which we call Drosophila IAP1 (DIAP1). Overexpression of DIAP1 or a related protein, DIAP2, in the eye suppresses normally occurring cell death as well as death due to overexpression of rpr or head involution defective. IAP death-preventing activity localizes to the N-terminal baculovirus IAP repeats, a motif found in both viral and cellular proteins associated with death prevention.

Amino Acid Sequence↗

KSR, a novel protein kinase required for RAS signal transduction.

We have identified and characterized two genes in Drosophila whose products are required for activated RAS to signal with normal efficiency, but do not appear to effect signaling by activated RAF. One encodes the beta subunit of type I geranylgeranyl transferase, a prenylation enzyme essential for targeting RAS to the plasma membrane. The other encodes a protein kinase that we have named kinase suppressor of ras (ksr). By genetic criteria, we show that KSR functions in multiple receptor tyrosine kinase pathways. We have isolated mammalian homologs of KSR that, together with the Drosophila gene, define a novel class of kinases. Our results suggest that KSR is a general and evolutionarily conserved component of the RAS signaling pathway that acts between RAS and RAF.

Amino Acid Sequence↗

hikaru genki, a CNS-specific gene identified by abnormal locomotion in Drosophila, encodes a novel type of protein.

We have identified a gene, hikaru genki (hig), whose mutant phenotype includes abnormal locomotor behavior. Mutant first instar larvae have uncoordinated movements, and both larvae and adults have reduced locomotion. Sequence analyses revealed that this gene encodes a novel type of protein with a signal sequence, but without transmembrane regions. One of its domains has similarities with immunoglobulin domains; three or four regions are similar to a complement-binding domain found in complement-related proteins and selectins. In situ hybridization to embryos revealed that accumulation of the hig transcripts is restricted to subsets of cells in the CNS. Our data suggest that hig has a role in the development of CNS functions involved in locomotor activity.

Amino Acid Sequence↗

iroquois: a prepattern gene that controls the formation of bristles on the thorax of Drosophila.

We have isolated a Drosophila mutant where the lateral parts of the notum are completely naked, leaving unaffected a median stripe of hairs. This mutation, iroquois (iro), defines a new gene which maps at 69D. We show that, in the presumptive lateral notum of mutant discs, sense organ precursor cells fail to form and the proneural gene scute is not expressed. The expression of a reporter gene inserted near iro suggests that iro itself is massively expressed in this region of the disc. We propose that iro is a prepattern gene essential to activate the expression of scute in the regions of the disc that will form the lateral notum.

Alleles↗

Genomic regulation of transposable elements in Drosophila.

Transposable elements are a major source of genetic change, including the creation of novel genes, the alteration of gene expression in development, and the genesis of major genomic rearrangements. They are ubiquitous among contemporary organisms and probably as old as life itself. The long coexistence of transposable elements in the genome would be expected to be accompanied by host-element coevolution. Indeed, the important role of host factors in the regulation of transposable elements has been illuminated by recent studies of several systems in Drosophila. These include host factors that regulate the P element, a host mutation that renders the genome permissive for gypsy mobilization and infection, and newly induced mutations that affect the expression of transposon insertion mutations. The finding of a type of hybrid dysgenesis in D. virilis, in which multiple unrelated transposable elements are mobilized simultaneously, may also be relevant to host-factor regulation of transposition.

Animals↗

Origins of sequence diversity in the malaria vaccine candidate merozoite surface protein-2 (MSP-2) in Amazonian isolates of Plasmodium falciparum.

The recent evolution of Plasmodium falciparum is at odds with the extensive polymorphism found in most genes coding for antigens. Here, we examined the patterns and putative mechanisms of sequence diversification in the merozoite surface protein-2 (MSP-2), a major malarial repetitive surface antigen. We compared the msp-2 gene sequences from closely related clones derived from sympatric parasite isolates from Brazilian Amazonia and used microsatellite typing to examine, in these same clones, the haplotype background of chromosome 2, where msp-2 is located. We found examples of msp-2 sequence rearrangements putatively created by nonreciprocal recombinational events, such as replication slippage and gene conversion, while maintaining the chromosome haplotype. We conclude that these nonreciprocal recombination events may represent a major source of antigenic diversity in MSP-2 in P. falciparum populations with low rates of classical meiotic recombination.

Alleles↗

Separation of sister chromatids in mitosis requires the Drosophila pimples product, a protein degraded after the metaphase/anaphase transition.

Mutations in the Drosophila genes pimples and three rows result in a defect of sister chromatid separation during mitosis. As a consequence, cytokinesis is also defective. However, cell cycle progression including the mitotic degradation of cyclins A and B is not blocked by the failure of sister chromatid separation, and as a result, metaphase chromosomes with twice the normal number of chromosome arms still connected in the centromeric region are observed in the following mitosis, pimples encodes a novel protein that is rapidly degraded in mitosis. Our observations suggest that Pimples and Three rows act during mitosis to release the cohesion between sister centromeres.

Amino Acid Sequence↗

Mutations in the heatshock cognate 70 protein (hsc4) modulate Notch signaling.

In our effort to dissect the Notch signaling mechanism we have conducted a screen for mutations that reduce Notch signaling activity. We recovered nine complementation groups as modifiers of the hypomorphic Notch allele notchoid. Apart from the known Notch signaling modulators Notch, Delta and mastermind we isolated alleles in vestigial, wingless, scalloped and clipped, genes known to affect wing morphogenesis. In addition, we identified mutations in Bag, the gene encoding clathrin heavy chain and a dominant mutation of the cytosolic 70 kDa heatshock cognate protein encoded by the hsc4 gene, as Notch signaling modifier. We focused our attention on the latter mutation because it displays dramatic genetic interactions with mutations of the Notch receptor as well as several additional Notch signaling pathway elements. We discuss how hsc4, a gene thought to be involved in subcellular trafficking, may affect the number of functional Notch receptors on the cell surface.

Alleles↗

Mediator, not holoenzyme, is directly recruited to the heat shock promoter by HSF upon heat shock.

Activators of RNA polymerase II (Pol II) transcription have been shown to bind several coactivators and basal factors in vitro. Whether such interactions play a primary regulatory role in recruiting these factors to activator-associated chromosomal target sites in living cells remains unclear. Here, we show that upon heat shock the Pol II-free form of Mediator is rapidly recruited to HSF binding sites. Unlike the TAFs and Pol II, the interaction between Mediator and HSF on chromosomal loci is direct and mechanistically separable from the preinitiation complex assembly step. Therefore, the activator-Mediator interaction likely underlies the initiation of signal transfer from enhancer-bound activators to the basal transcription machinery.

Amanitins↗

Genetic control of male cuticular hydrocarbons in Drosophila melanogaster.

7-tricosene (7-T) and 7-pentacosene (7-P) are the two main hydrocarbons on the cuticle of male Drosophila melanogaster. These two substances might play a pheromonal role during courtship behaviour. We investigated the genetic basis of the quantitative polymorphism observed in the production of 7-T and 7-P. Strains of different geographic origin, with males producing either predominantly 7-T or predominantly 7-P, were hybridized with strains carrying genetic markers. We found that chromosome II changes the balance between 7-T and 7-P while chromosome III regulates the overall quantity of both 7-monoenes. We have also characterized and roughly mapped sept and smoq, two genetic factors on chromosome II that act additively on the production of both cuticular hydrocarbons. The genetic control of the variation in 7-T and 7-P varies between D. melanogaster strains and between D. melanogaster and its sibling species D. simulans. The possible evolutionary and physiological causes of this variation as well as its functional implication for courtship behaviour are discussed.

Alkenes↗

Rates of movement and distribution of transposable elements in Drosophila melanogaster: in situ hybridization vs Southern blotting data.

Genomic copy numbers and the rates of movement of nine families of transposable elements (TEs) of Drosophila melanogaster were estimated in two sets of mutation accumulation lines: Beltsville and Madrid. Southern blotting was used to screen a large number of samples from both genetic backgrounds for TEs. The Madrid lines were also screened by in situ hybridization of TEs to polytene chromosomes, in order to obtain more detailed information about the behaviour of TEs in the euchromatin. Southern blotting data provided evidence of insertions and excision events in both genetic backgrounds, occurring at rates of approximately 10(-5) and 10(-6) per element copy per generation, respectively. In contrast, in situ data from the Madrid background presented a completely different picture, with no evidence for excisions, and a significantly higher rate of transposition (1.01 x 10(-4)). Direct comparison of the two data sets suggests that the Southern blotting technique had serious deficiencies: (i) it underestimated element abundance; (ii) it revealed less than 30% of the new insertions detected by in situ hybridization; and (iii) changes in the size of restriction fragments from any source were spuriously identified as simultaneous insertion-excision events. Our in situ data are consistent with previous studies, and suggest that selection is the main force controlling element spread by transposition.

Animals↗

A QTL analysis of female variation contributing to refractoriness and sperm competition in Drosophila melanogaster.

Sperm competition is an important fitness component in many animal groups. Drosophila melanogaster males exhibit substantial genetic variation for sperm competitive ability and females show considerable genetic variation for first versus second male sperm use. Currently, the forces responsible for maintaining genetic variation in sperm competition related phenotypes are receiving much attention. While several candidate genes contributing to the variation seen in male competitive ability are known, genes involved in female sperm use remain largely undiscovered. Without knowledge of the underlying genes, it will be difficult to distinguish between different models of sexual selection such as cryptic female choice and sexual conflict. We used quantitative trait locus (QTL) mapping to identify regions of the genome contributing to female propensity to use first or second male sperm, female refractoriness to re-mating, and early-life fertility. The most well supported markers influencing the phenotypes include 33F/34A (P2), 57B (refractoriness) and 23F/24A (fertility). Between 10% and 15% of the phenotypic variance observed in these recombinant inbred lines was explained by these individual QTLs. More detailed investigation of the regions detected in this experiment may lead to the identification of genes responsible for the QTLs identified here.

Animals↗

Chromosomal responses of blowfly Lucilia cuprina to heat and heavy metal stress.

Chromosomal responses to heat and heavy metal shocks were studied in the pupal trichogen polytene chromosomes of Lucilia cuprina. Heat shock induced seven distinct puffs on different chromosomes at the following loci: 1C2, 21A1, 21C, 23B, 24B, 42A and 95B3. Arsenate and mercury, two of the most common toxic environmental chemical pollutants also, induced almost the same set of puffs except 20B2, which appeared to be induced by metals only and 95B3, which was not induced by arsenate. The findings suggest that a common set of gene loci encoding the heat shock proteins is responsive to these diverse environmental stresses.

Animals↗

Characterization of the Drosophila melanogaster genome at the nuclear lamina.

The nuclear lamina binds chromatin in vitro and is thought to function in its organization, but genes that interact with it are unknown. Using an in vivo approach, we identified approximately 500 Drosophila melanogaster genes that interact with B-type lamin (Lam). These genes are transcriptionally silent and late replicating, lack active histone marks and are widely spaced. These factors collectively predict lamin binding behavior, indicating that the nuclear lamina integrates variant and invariant chromatin features. Consistently, proximity of genomic regions to the nuclear lamina is partly conserved between cell types, and induction of gene expression or active histone marks reduces Lam binding. Lam target genes cluster in the genome, and these clusters are coordinately expressed during development. This genome-wide analysis gives clear insight into the nature and dynamic behavior of the genome at the nuclear lamina, and implies that intergenic DNA functions in the global organization of chromatin in the nucleus.

Animals↗

Genomic and functional evolution of the Drosophila melanogaster sperm proteome.

In addition to delivering a haploid genome to the egg, sperm have additional critical functions, including egg activation, origination of the zygote centrosome and delivery of paternal factors. Despite this, existing knowledge of the molecular basis of sperm form and function is limited. We used whole-sperm mass spectrometry to identify 381 proteins of the Drosophila melanogaster sperm proteome (DmSP). This approach identified mitochondrial, metabolic and cytoskeletal proteins, in addition to several new functional categories. We also observed nonrandom genomic clustering of sperm genes and underrepresentation on the X chromosome. Identification of widespread functional constraint on the proteome indicates that sexual selection has had a limited role in the overall evolution of D. melanogaster sperm. The relevance of the DmSP to the study of mammalian sperm function and fertilization mechanisms is demonstrated by the identification of substantial homology between the DmSP and proteins of the mouse axoneme accessory structure.

Animals↗

Cytogenetic mechanism and genetic consequences of thelytoky in the wasp Trichogramma cacoeciae.

In Hymenoptera, complete parthenogenesis, that is thelytoky, is a common phenomenon where virgin females produce only daughters. Thelytoky is often induced by bacteria of the genus Wolbachia, but can also be genetically determined by the insect itself, as in the genus Trichogramma where both forms exist. In order to compare these two forms of thelytoky, chromosome behaviour analysis in young eggs and genetic analysis of microsatellite markers were carried out in the wasp Trichogramma cacoeciae, where thelytoky is genetically determined. Microscopic studies revealed that during female gamete formation meiotic cells undergo only a single equational division followed by the expulsion of a single polar body. This absence of meiotic recombination and reduction corresponds well with the high levels of heterozygosity observed in females collected from the field and a nonsegregation pattern in the offspring of heterozygous females. We therefore concluded that diploidy in T. cacoeciae is maintained through an apomictic cloning mechanism and that the incidence of thelytoky under genetic control of the wasp differs entirely from the mechanism induced by Wolbachia infection, where thelytoky is restored through gamete duplication.

Animals↗