Y-chromosome dimorphism in the malaria vector Anopheles stephensi from south India.
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The way that the malaria vectors Anopheles arabiensis and An.gambiae survive the dry season in sub-saharan Sahel or northern Savanna areas of Africa remains enigmatic. We examined this problem by calculating the effective sizes (Ne) of An.arabiensis populations for several locations in West Africa. An indirect/genetic procedure was used, comparing gene frequencies at several time intervals. The amount of drift which occurred provides an estimate of Ne. Most estimates of Ne were approximately 2000 individuals, probably close to the yearly minimum. This supports the hypothesis that populations of An.arabiensis in this region are continuous throughout the year, with many individuals surviving through the dry season, perhaps in a physiologically altered state, rather than extinction or severe bottlenecks during the dry season, followed by recolonization by a few individual survivors or immigrants in the subsequent rainy season.
Cross-species hybridization has been used to isolate a second Drosophila gene, with homology to a feline glutamate decarboxylase (Gad) cDNA. The gene differs in sequence, chromosomal location, and spatial expression from the previously reported Drosophila Gad gene, but both encode proteins of 58 kDa. The derived amino acid sequence reveals a typical pyridoxal phosphate binding site and sequence homology consistent with a glutamate decarboxylase function. The protein includes an amino-terminal polyasparagine sequence, and a beta-pleated sheet region, with regularly spaced glutamine and arginine residues, not found in other decarboxylases. Expression in the adult is limited to the neuropil of the first optic ganglion and to regions of the thoracic musculature that may correspond to the location of motor neuron axons. This is consistent with a glial localization for the transcript. There is no overlap with the reported expression of Drosophila Gad. Although the molecular evidence suggests that this gene encodes a pyridoxal phosphate-dependent decarboxylase, glutamate decarboxylase activity associated with this gene could not be demonstrated, and the in vivo substrate is unknown. It is possible that the protein encoded by this gene is novel, not only in sequence and spatial expression, but also in substrate specificity.
D. subobscura, a typically Palearctic species, has recently colonized the American continent. The population from which the colonization started is still unknown. A complete association between a lethal gene and the O5 chromosomal inversion has been detected in the colonizing populations; this association could be used to ascertain the origin of the colonization. In the present study, three hypotheses on the origin of this association are analyzed and their probabilities are calculated. According to the first hypothesis an O5 chromosome carrying a lethal gene would have been included in the sample from which the colonization started, thus originating the association found in the American populations. The second hypothesis assumes that, at the beginning of the colonization, the association between the O5 inversion and a lethal gene would have been formed from an Ost chromosome carrying a lethal gene, on which a new O5 (or similar) inversion arose. In the last hypothesis, the association between the O5 inversion and the lethal gene would have originated at the beginning of the colonization from a lethal-free O5 chromosome, in which a lethal gene arose due to mutation.
Successful propagation of the malaria parasite Plasmodium falciparum within a susceptible mosquito vector is a prerequisite for the transmission of malaria. A field-based genetic analysis of the major human malaria vector, Anopheles gambiae, has revealed natural factors that reduce the transmission of P. falciparum. Differences in P. falciparum oocyst numbers between mosquito isofemale families fed on the same infected blood indicated a large genetic component affecting resistance to the parasite, and genome-wide scanning in pedigrees of wild mosquitoes detected segregating resistance alleles. The apparently high natural frequency of resistance alleles suggests that malaria parasites (or a similar pathogen) exert a significant selective pressure on vector populations.
We surveyed an Anopheles gambiae population in a West African malaria transmission zone for naturally occurring genetic loci that control mosquito infection with the human malaria parasite, Plasmodium falciparum. The strongest Plasmodium resistance loci cluster in a small region of chromosome 2L and each locus explains at least 89% of parasite-free mosquitoes in independent pedigrees. Together, the clustered loci form a genomic Plasmodium-resistance island that explains most of the genetic variation for malaria parasite infection of mosquitoes in nature. Among the candidate genes in this chromosome region, RNA interference knockdown assays confirm a role in Plasmodium resistance for Anopheles Plasmodium-responsive leucine-rich repeat 1 (APL1), encoding a leucine-rich repeat protein that is similar to molecules involved in natural pathogen resistance mechanisms in plants and mammals.
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The gene encoding the major capsid protein of the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) was identified, sequenced, and transcriptionally mapped. The location of the gene was determined by immunological screening of an expression library of AcMNPV open reading frame-beta-galactosidase fusions with an antibody raised to virus structural proteins. The DNA sequence of the corresponding region, which mapped within 56.6 and 58.0 map units on the AcMNPV genome, revealed a 1,040-base-pair open reading frame capable of encoding a 39-kilodalton polypeptide. The identity of the polypeptide was determined by Western blot (immunoblot) analysis of purified empty capsids with an antibody raised to the capsid-beta-galactosidase fusion protein. The identity of the peptide encoded by the gene was confirmed by immunoprecipitation of an in vitro translation product with RNA selected by hybridization to DNA sequences from the coding region of the gene. Transcripts of the capsid gene were analyzed by Northern (RNA) blots and mapped by nuclease protection and primer extension analysis. The capsid gene is transcribed maximally at 12 and 24 h postinfection but not in the presence of cycloheximide, a protein synthesis inhibitor, or aphidicolin, a viral DNA synthesis inhibitor, and is therefore classified as a late gene. The gene is transcribed in a counterclockwise direction with respect to the circular map. There are three transcriptional start sites, all containing the AGTAAG consensus sequence found at the start site of all late AcMNPV genes.
Spatial organization of chromatin in the interphase nucleus plays a role in gene expression and inheritance. Although it appears not to be random, the principles of this organization are largely unknown. In this work, we show an explicit relationship between the intranuclear localization of various chromosome segments and the pattern of gene distribution along the genome sequence. Using a 7-megabase-long region of the Drosophila melanogaster chromosome 2 as a model, we observed that the six gene-poor chromosome segments identified in the region interact with components of the nuclear matrix to form a compact stable cluster. The six gene-rich segments form a spatially segregated unstable cluster dependent on nonmatrix nuclear proteins. The resulting composite structure formed by clusters of gene-rich and gene-poor regions is reproducible between the nuclei. We suggest that certain aspects of chromosome folding in interphase are predetermined and can be inferred through in silico analysis of chromosome sequence, using gene density profile as a manifestation of "folding code."
In this review, we combine the results of our published and unpublished work with the published results of other laboratories to provide an updated map of the centromeric heterochromatin of chromosome 3 in Drosophila melanogaster. To date, we can identify more than 20 genes (defined DNA sequences with well-characterized functions and (or) defined genetic complementation groups), including at least 16 essential loci. With the ongoing emergence of data from genetic, cytological, and genome sequencing studies, we anticipate continued, substantial progress towards understanding the function, structure, and evolution of centric heterochromatin.
Aiming to establish phylogenetic relationships among species of the montium subgroup, detailed polytene chromosome maps are given showing intraspecific polymorphism and ecdysone induced larval puffing pattern profiles of two Afrotropical members of this subgroup, Drosophila diplacantha and D. seguyi. Both species exhibit two unique characteristics that define the montium subgroup, namely, a large number of reverse tandem duplications and a progressive darkening of anterior spiracles of the late third instar larvae, which is accompanied by a definite temporal and spatial puffing pattern of the salivary gland chromosomes. In contrast with the well-formed Balbiani ring 1 (BR1) observed in Oriental and Indian montium species, BR1 exhibits a different developmental profile in D. diplacantha, while it is obscured in D. seguyi. Although phyletic comparisons of five species from five different complexes within the subgroup show some conservation in banding and puffing pattern homologies, an analysis to assign map sections by sequential rearrangements remains unresolved at this time. The evolution of the subgroup is discussed in relation with the sharing of reverse tandem duplications, especially those including the montium BRs.
The Simulium neavei group is medically important as a vector of human onchocerciasis and is unique among blackflies because of the attachment to freshwater crabs during parts of the larval and pupal life cycle stages. Detailed larval salivary gland polytene chromosome maps are presented for two taxa designated S. neavei Amani form A and S. neavei Amani form B, which are probably synonymous with the previously described morphospecies, S. nyasalandicum and S. woodi, respectively. Simulium neavei Amani form B differs from S. neavei Amani form A by three paracentric inversions, a large deletion and a chromocentre, while the latter taxon differs from the former one by a mid-arm interchange. The fixed mid-arm interchange (1-tr) between two related taxa is unique among the Simuliidae. Detailed banding analysis of S. neavei Amani form B shows that, relative to S. neavei Amani form A, sections 16a, 16b, and 17c of the short arm of chromosome one (IS) have been inserted into section 69/70 of the short arm of chromosome three (IIIS). Assignment of directionality for this interchange inferred because of the near universality in the Simuliidae of a chromosome III long arm to short arm ratio of 2:1. We propose that the mid-arm interchange acted as a primary isolating mechanism because of the segregational load on F1 heterozygous interchange progeny. The magnitude of the segregational load would be dependent on the frequency of pairing and crossing-over between the transposed interstitial segment and its original counterpart in the chromosome complement. Nonpairing in the transposed region and normal random segregation would presumably lead to a 50% of gametes carrying the duplicated or deficient transposed segment, resulting in a segregation load of 50%. However, crossing-over in the paired interstitial region may greatly increase segregational load as result of multivalent formation at metaphase one (MI) and the generation of dicentric bridge(s) and acentric fragment(s) upon cell division. Such meiotic disjunction irregularities could lead to meiotic arrest, gametic loss and (or) cell restitution, and the eventual formation of polyploid (lethal) zygotes.
Human onchocerciasis is hyper-endemic in south-western Cameroon and the island of Bioko. The vector on the island is the endemic 'Bioko' form of Simulium yahense. On the mainland, S. damnosum s.s., S. squamosum and S. mengense are recorded around Mount Cameroon but their contribution to transmission in the area is unclear. As elimination of onchocerciasis, through vector eradication, is a possibility on the island, it is important to be able to identify vector flies which might migrate to Bioko from the mainland. The morphologies of S. damnosum s.s. and S. squamosum are already well known. New cytotaxonomic and morphotaxonomic descriptions of S. mengense, a species which is less well known, are presented. Simulium mengense can be distinguished from the other cytospecies in the area by the presence of tufts of hair-like setae on the larval thorax, the presence of hairs on the subcostal vein of the adult female, and by the scutal pattern of the adult male. Although Vajime and Dunbar described seven fixed inversions in S. mengense, in 1977, nine were observed in the present study. Of the nine, three were the same as Vajime and Dunbar's but the other six were either missed or misinterpreted by them.
Neural receptor-linked protein tyrosine phosphatases (RPTPs) are required for guidance of motoneuron and photoreceptor growth cones in Drosophila. These phosphatases have not been implicated in growth cone responses to specific guidance cues, however, so it is unknown which aspects of axonal pathfinding are controlled by their activities. Three RPTPs, known as DLAR, DPTP69D, and DPTP99A, have been genetically characterized thus far. Here we report the isolation of mutations in the fourth neural RPTP, DPTP10D. The analysis of double mutant phenotypes shows that DPTP10D and DPTP69D are necessary for repulsion of growth cones from the midline of the embryonic central nervous system. Repulsion is thought to be triggered by binding of the secreted protein Slit, which is expressed by midline glia, to Roundabout (Robo) receptors on growth cones. Robo repulsion is downregulated by the Commissureless (Comm) protein, allowing axons to cross the midline. Here we show that the Rptp mutations genetically interact with robo, slit and comm. The nature of these interactions suggests that DPTP10D and DPTP69D are positive regulators of Slit/Roundabout repulsive signaling. We also show that elimination of all four neural RPTPs converts most noncrossing longitudinal pathways into commissures that cross the midline, indicating that tyrosine phosphorylation controls the manner in which growth cones respond to midline signals.
Cytological investigations of salivary gland polytene chromosomes of the Drosophila kikkawai complex in Thailand have revealed an interesting pattern of species divergence. Drosophila bocki, D. leontia and D. kikkawai share chromosome arrangement 3LB. However, additional gene arrangements of 3LA and 3LC have been observed in D. kikkawai and D. leontia, respectively, while D. bocki remains polymorphic for inversion 3LB. Chromosomal evidence seems to suggest that D. bocki is similar to a common ancestor of the D. kikkawai complex.
Curly is a easily distinguishable dominant mutant wing character. The Cy mutation is the most commonly used dominant marker for the second chromosome balancers in Drosophila melanogaster, but little is known about the Cy gene. Based on known genomic and cytogenetic information, a 102 bp deletion which is located between the Genes synaptotagmin (syt) and Activin Like Protein at 23B(Alp23B) on the Drosophila melanogaster genome (release 4) had been found to be commonly contained on Cy chromosome in three different curly strains. Meanwhile, when using the deletion as a DNA marker, the result suggested that Cy homozygote be lethal in embryo period. These results will provide some helpful information to investigate molecular mechanism of curly wings in the further study.
The formation of the Drosophila wing involves developmental processes such as cell proliferation, pattern formation, and cell differentiation that are common to all multicellular organisms. The genes controlling these cellular behaviors are conserved throughout the animal kingdom, and the genetic analysis of wing development has been instrumental in their identification and functional characterization. The wing is a postembryonic structure, and most loss-of-function mutations are lethal in homozygous flies before metamorphosis. In this manner, loss-of-function genetic screens aiming to identify genes affecting wing formation have not been systematically utilized. As an alternative, a number of genetic searches have utilized the phenotypic consequences of gene gain-of-expression, as a method more efficient to search for genes required during imaginal development. Here we present the results of a gain-of-function screen designed to identify genes involved in the formation of the wing veins. We generated 13,000 P-GS insertions of a P element containing UAS sequences (P-GS) and combined them with a Gal4 driver expressed mainly in the developing pupal veins. We selected 500 P-GSs that, in combination with the Gal4 driver, result in modifications of the veins, changes in the morphology of the wing, or defects in the differentiation of the trichomes. The P-element insertion sites were mapped to the genomic sequence, identifying 373 gene candidates to participate in wing morphogenesis and vein formation.