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The non-venom insect phospholipases A2.

Phospholipases A(2) (PLA(2)s) are responsible for releasing the fatty acid moiety from the sn-2 position of phospholipids. These enzymes are virtually ubiquitous proteins known from all major biological taxa. Various PLA(2)s act in a wide array of biological processes, including digestion of dietary lipids, cellular homeostasis, intra- and intercellular signaling, host defense and at least a few ecological interactions. PLA(2) activities have been recorded in a small number of insect species, which can be taken to represent the broad group, Insecta. Within insects, PLA(2)s act in functions expected from the background on these enzymes. So far, we know PLA(2)s act in lipid digestion, cellular host defense signaling, reproduction and in organismal-level metabolism. Additional PLA(2) actions are certain to emerge. This is the first article devoted to assembling the known information on insect PLA(2)s. I review the scant information available on the biological actions of PLA(2)s in insects, relate new findings on insect pathogens that disrupt insect immune functions by inhibiting PLA(2)s and mention the few reports of sequence information on insect PLA(2)s. Finally, I offer a brief prospectus on future research into insect PLA(2)s. There are two overarching points in this paper. One, there remains a great deal to learn about insect PLA(2)s and two, some of the findings on insect PLA(2)s will have meaningful practical significance.

Animals↗

Virus-derived genes for insect-resistant transgenic plants.

Insect viruses have evolved to counter physiological barriers to infection presented by the host insect. For the Lepidoptera (butterflies and moths), these barriers include (1) the peritrophic membrane (PM) lining the gut, which presents a physical barrier to virus infection of the midgut epithelial cells, (2) the basement membrane (BM) that overlies the gut thereby restricting secondary infection of other tissues, and (3) the immune system of the host insect. Hence, insect viruses provide a resource for genes that disrupt host physiology in a specific manner, and these genes in turn serve as a resource both for the study of physiological processes, and for disruption of these processes for pest management purposes. There are several examples of the application of genes used by an insect virus to overcome the PM barrier for production of insect-resistant transgenic plants. There are other examples of intrahemocoelic effectors, such as BM-degrading proteases that can only be used with an appropriate system for delivery of the agent from the gut into the hemocoel (body cavity) of the insect pest. In this chapter, we describe (1) baculovirus- and entomopoxvirus-derived genes that alter the physiology of the host insect, (2) use of these and homologous genes for production of insect-resistant transgenic plants, (3) other viral genes that have potential for use in development of insect-resistant transgenic plants, and (4) the use of plant lectins for delivery of intrahemocoelic toxins from transgenic plants. Plant expression of polydnavirus-derived genes is described by Gill et al. (this volume, pp. 393-426).

Animals↗

The effect of sunscreen on the efficacy of insect repellent: a clinical trial.

Sunscreen loses efficacy when used with the insect repellent N,N -diethyl-3-methylbenzamide (diethyltoluamide; deet). A previous study demonstrated a 33.3% decrease in sun protection factor when insect repellent and sunscreen were applied sequentially to the skin. The purpose of this study was to determine whether the efficacy of insect repellent is affected by two different formulations of sunscreen. The combination of sunscreen and insect repellent was tested in 8 groups: a control group (no sunscreen or insect repellent), sunscreen alone (two groups [gel and cream]), insect repellent alone, insect repellent applied before sunscreen (two groups), and insect repellent applied after sunscreen (two groups). The results of this study showed that insect repellent has the same efficacy even when sunscreen is applied with it. This indicates that in the formulations tested, sunscreen does not reduce the efficacy of insect repellent.

Drug Combinations↗

Detection of stored-grain insect infestation in wheat transported in railroad hopper-cars.

Levels of insect infestation, insect spatial distribution, and the relationship between the number of insect-damaged kernels (IDK) and the number of insects present in grain samples in three-hopper railcars transporting wheat from country elevators to a mill were studied. Six of eight sampled railcars were infested with more than two species of insects. The most abundant species collected were the lesser grain borer, Rhyzopertha dominica (F.), and rusty grain beetle, Cryptolestes ferrugineus (Stephens), with the larval stage of the two species being the most prevalent (>90%). The spatial distributions of these two species within the grain mass were typically clumped in railcar compartments containing >0.4 insect/2.75-kg sample of wheat, and these foci of high-infestation levels varied in compartments within the railcars and among the sampled railcars. There were no significant correlations between IDK and insect density for any of the different stage-specific insect populations that were collected in the grain samples. Mean numbers of immatures and IDK differed among railcars and compartments within railcars, but not among grain depths. Number of insects in the first discharge sample was not correlated with mean numbers of insects in the entire compartment. This indicates that each compartment of a railcar should be sampled to determine level of insect infestation but that sampling at different depths within a compartment is less important.

Animals↗

Pest insect control in organically-produced crops of field vegetables.

In the UK, the demand for organic vegetable and salad crops is increasing, mainly as a result of the requirements of the multiple retailers. However, approximately 85% of the organic fruit and vegetable produce sold in the UK is imported. A major constraint to growing field vegetable crops, and particularly organically-produced crops, is the reduction in crop yield and quality caused by pest insects. This paper will consider the control techniques currently available to organic growers and other techniques that may become available in the future. Growing plant varieties with complete or even partial resistance to pest insects can be an effective way of reducing crop damage. There are already varieties of carrot, with resistance to carrot fly, and lettuce, with resistance to certain pest aphid species, which are available commercially. Cultural techniques to exclude, deter or avoid pest insects are also being used by some organic growers. Although isolating new crops from sources of infestation can be a highly effective control strategy, many organic growers cannot use it, as the land converted for organic production is still limited. Various crop covers can be used to prevent pest insects from damaging field crops, but to be effective such covers have to be in place before the pests enter the crop. Several researchers have tried to develop techniques to prevent pest insects from finding their host-plants. No technique involving semiochemicals has been sufficiently successful to be used in field vegetable production in the UK. Other studies have shown that the numbers of pest insects found on crop plants are reduced considerably when the crop is allowed to become weedy, is intercropped with another plant species, or is undersown with a living mulch. Hence, work is now needed to select background plant species that will both reduce pest insect numbers and cause the least reduction in yield to the harvested crop plants. There is also a need to obtain a better understanding of "companion planting", a practice used frequently by organic growers. To date, microbial control is the only biological technique that has been used successfully in field vegetable crops in the UK. However, only the toxicant produced by one microbial agent, the bacterium Bacillus thuringiensis, has so far been registered for use. The use of bacteria, fungi and viruses to control pests of field vegetable crops certainly has possibilities. However, in many cases there are still problems to be overcome to select pathogens that are compatible with, or can still be effective in, the wide fluctuations in temperature, humidity and soil moisture that occur under field conditions. Attempts are now being made to use entomopathogenic nematodes and predatory arthropods to control one major pest insect, the cabbage root fly. Techniques developed to improve the timing of application of various crop protection procedures in systems of conventional vegetable production apply equally well to organic production, despite the choice of control options being more limited. In particular, models to forecast the timing of pest insect attacks could be used to great effect, to indicate the best times to plant, protect and harvest a specific crop to minimise pest insect damage.

Agriculture↗

New north american records of aquatic insects as paratenic hosts of pheromermis (Nematoda: mermithidae)

Several species of aquatic insects in Trout Park Nature Preserve (Elgin, IL) were observed to have small, black spots (<0.1 mm diameter) visible within their bodies. Microscopic examination revealed these spots to be coiled juveniles of a mermithid (Nematoda: Mermithidae). Based on host habitat (seepage areas and rivulets), host species (aquatic insects), and size (mean diameter of coiled juveniles = 79 &mgr;m), it is likely that these mermithids were in the genus Pheromermis. Since adult mermithids were not found, species determination was not feasible, and the possibility of a new species cannot be ruled out. Pheromermis pachysoma and Pheromermis vesparum, however, are two species known to use aquatic insects as paratenic (i.e., transport) hosts in order to reach their definitive hosts, vespid wasps. Wasp larvae are infected by consuming the flesh of adult aquatic insects that contain the coiled juveniles of these Pheromermis spp. Of the 19 macroinvertebrate species examined in this study, Pheromermis juveniles were found in 4 caddisfly species (Hesperophylax designatus, Lepidostoma liba, Glossosoma intermedium, and Diplectrona modesta) and in 2 stonefly species (Clioperla clio and Amphinemura delosa). In addition to all 6 insect species being new host records for Pheromermis infection, this also represents the first report of nematode infection in stoneflies within the Western Hemisphere and of a Pheromermis sp. in Illinois. Among trophic groups, insect detritivores have been frequently recorded infected with coiled Pheromermis juveniles because of their direct consumption of eggs, and we also observed this for detritivores in our investigation (e.g., L. liba and A. delosa). Because C. clio was intensively infected, however, our study also provided evidence that predatory insects can be paratenic hosts. Coiled juveniles were typically present in muscle and fat body and present in almost all body regions. Not every infected paratenic host had external signs of infection; thus, dissections are required for accurate determination of infection prevalence and intensity. Our findings, in conjunction with those made in previous studies, indicate that a wide variety of aquatic insects may be used as paratenic hosts by Pheromermis. Because of their small size, it is highly likely that coiled juveniles are either overlooked or mistaken for melanized integumental wounds during field studies of aquatic insects. A more careful inspection for these coiled juveniles in aquatic insects, especially detritivores and their predators in seepage areas, would probably reveal that Pheromermis is far more common than presently documented. Copyright 1999 Academic Press.

Journal Article↗

GABA receptor molecules of insects.

Receptors for 4-aminobutyric acid (GABA) have been identified in both central and peripheral nervous systems of several invertebrate phyla. To date, much of the information derived from physiological and biochemical studies on insect GABA receptors relates to GABA-gated chloride channels that show some similarities with vertebrate GABAA receptors. Like their vertebrate central nervous system (CNS) counterparts, agonist activation of such insect GABA receptors leads to a rapid, picrotoxin-sensitive increase in chloride ion conductance across the cell membrane. In insects, responses to GABA can be modulated by certain benzodiazepines and barbiturates. However, recent studies have detected a number of striking pharmacological differences between GABA-gated chloride channels of insects and vertebrates. Receptor binding, electrophysiological and 36Cl- flux assays have indicated that many insect receptors of this type are insensitive to the vertebrate GABAA antagonists bicuculline and pitrazepin. Benzodiazepine binding sites coupled to insect GABA receptors display a pharmacological profile distinct from that of corresponding sites in vertebrate CNS. Receptor binding studies have also demonstrated differences between convulsant binding sites of insect and vertebrate receptors. Insect GABA receptor molecules are important target sites for several chemically-distinct classes of insecticidally-active molecules. By characterizing these pharmacological properties in detail, it may prove possible to exploit differences between vertebrate and insect GABA receptors in the rational design of novel, more selective pest control agents. The recent application of the powerful techniques of molecular biology has revealed a diversity of vertebrate GABAA receptor subunits and their respective isoforms that can assemble in vivo to form a multiplicity of receptor subtypes. Molecular cloning of insect GABA receptor subunits will not only enhance our understanding of invertebrate neurotransmitter receptor diversity but will also permit the precise identification of the sites of action of pest control agents.

Amino Acid Sequence↗

Pathogenicity of mollicutes for insects: possible use in biological control.

Acholeplasmas, spiroplasmas and other non-helical sterol-requiring mycoplasmas of unknown phylogenetic affinity inhabit insects. Of these, only spiroplasmas are known to be pathogenic. Group I-2 spiroplasmas, or Spiroplasma apis, especially in combination with other organisms, reduce honey-bee longevity. Plant pathogenic mycoplasma-like organisms are often found intracellularly in insects. Spiroplasmas are found predominantly in the gut lumen or haemolymph (or both) of their insect hosts. Pathogenicity of mycoplasmas is usually altered by extended passage in unusual hosts, in only one of two alternate hosts, or in culture media. Enhancement of experimental pathogenicity may occur with extended cultural passages, but maintenance of natural pathogenicity must be accomplished by continuous exposure to the usual host. Recent data provide new information on the ecology of pathogenicity. Spiroplasmas from unique habitats also tend to be unique. Spiroplasmas isolated from flowers appear to be adapted to insect species that frequent floral surfaces. Group IV spiroplasmas have been isolated from members of 4 holometabolous insect orders (including Lepidoptera), all of which visit flowers. Social or predatory insects, or insects with an "aggregation" phase in their life histories, also appear to be prone to spiroplasma infection. Some insect species which harbor spiroplasmas also carry infections of other mollicutes, some of which involve the haemolymph. Appearance of spiroplasmas in adult insects in nature is strongly affected by seasonality. Extensive tests of the host ranges of the new insect mollicutes will be required before their suitability for biological control can be evaluated.

Acholeplasma↗

Climatic variability and the evolution of insect freeze tolerance.

Insects may survive subzero temperatures by two general strategies: Freeze-tolerant insects withstand the formation of internal ice, while freeze-avoiding insects die upon freezing. While it is widely recognized that these represent alternative strategies to survive low temperatures, and mechanistic understanding of the physical and molecular process of cold tolerance are becoming well elucidated, the reasons why one strategy or the other is adopted remain unclear. Freeze avoidance is clearly basal within the arthropod lineages, and it seems that freeze tolerance has evolved convergently at least six times among the insects (in the Blattaria, Orthoptera, Coleoptera, Hymenoptera, Diptera and Lepidoptera). Of the pterygote insect species whose cold-tolerance strategy has been reported in the literature, 29% (69 of 241 species studied) of those in the Northern Hemisphere, whereas 85 % (11 of 13 species) in the Southern Hemisphere exhibit freeze tolerance. A randomization test indicates that this predominance of freeze tolerance in the Southern Hemisphere is too great to be due to chance, and there is no evidence of a recent publication bias in favour of new reports of freeze-tolerant species. We conclude from this that the specific nature of cold insect habitats in the Southern Hemisphere, which are characterized by oceanic influence and climate variability must lead to strong selection in favour of freeze tolerance in this hemisphere. We envisage two main scenarios where it would prove advantageous for insects to be freeze tolerant. In the first, characteristic of cold continental habitats of the Northern Hemisphere, freeze tolerance allows insects to survive very low temperatures for long periods of time, and to avoid desiccation. These responses tend to be strongly seasonal, and insects in these habitats are only freeze tolerant for the overwintering period. By contrast, in mild and unpredictable environments, characteristic of habitats influenced by the Southern Ocean, freeze tolerance allows insects which habitually have ice nucleators in their guts to survive summer cold snaps, and to take advantage of mild winter periods without the need for extensive seasonal cold hardening. Thus, we conclude that the climates of the two hemispheres have led to the parallel evolution of freeze tolerance for very different reasons, and that this hemispheric difference is symptomatic of many wide-scale disparities in Northern and Southern ecological processes.

Acclimatization↗

Insect resistant cotton plants.

We have expressed truncated forms of the insect control protein genes of Bacillus thuringiensis var. kurstaki HD-1(cryIA(b) and HD-73 (cryIA(c) in cotton plants at levels that provided effective control of agronomically important lepidopteran insect pests. Total protection from insect damage of leaf tissue from these plants was observed in laboratory assays when tested with two lepidopteran insects, an insect relatively sensitive to the B.t.k. insect control protein, Trichoplusia ni (cabbage looper) and an insect that is 100 fold less sensitive, Spodoptera exigua (beet armyworm). Whole plants, assayed under conditions of high insect pressure with Heliothis zea (cotton bollworm) showed effective square and boll protection. Immunological analysis of the cotton plants indicated that the insect control protein represented 0.05% to 0.1% of the total soluble protein. We view these results as a major step towards the agricultural use of genetically modified plants with insect resistance in this valuable, high acreage crop.

Amino Acid Sequence↗

Habitat association in two genetic groups of the insect-pathogenic fungus metarhizium anisopliae: uncovering cryptic species?

Strains of insect-pathogenic fungi with high virulence toward certain pest insects have great potential for commercial biological control applications. Identifying such strains has been a central theme in using fungi for biological control. This theme is supported by a persistent paradigm in insect pathology which suggests that the host insect is the predominant influence on the population genetics of insect-pathogenic fungi. In this study, a population genetics analysis of the insect-pathogenic fungus Metarhizium anisopliae from forested and agricultural habitats in Ontario, Canada, showed a nonrandom association of alleles between two distinct, reproductively isolated groups (index of multilocus association = 1.2). Analyses of the mitochondrial DNA showed no differences between the groups. The two groups were associated with different habitat types, and associations with insect hosts were not found. The group from forested areas showed an ability for cold-active growth (i.e., 8 degrees C), while the group from the agricultural area showed an ability for growth at high temperatures (i.e., 37 degrees C) and resilience to UV exposure. These results represent a significant paradigm shift; habitat selection, not host insect selection, drives the population structure of these insect-pathogenic deuteromycetous fungi. With each group we observed recombining population structures as well as clonally reproducing lineages. We discuss whether these groups may represent cryptic species. Worldwide, M. anisopliae may be an assembly of cryptic species, each adapted to certain environmental conditions. The association of fungal genotypes with habitat but not with host insects has implications on the criteria for utility of this, and perhaps other, fungal biocontrol agents.

Agriculture↗

Stages of infection during the tripartite interaction between Xenorhabdus nematophila, its nematode vector, and insect hosts.

Bacteria of the genus Xenorhabdus are mutually associated with entomopathogenic nematodes of the genus Steinernema and are pathogenic to a broad spectrum of insects. The nematodes act as vectors, transmitting the bacteria to insect larvae, which die within a few days of infection. We characterized the early stages of bacterial infection in the insects by constructing a constitutive green fluorescent protein (GFP)-labeled Xenorhabdus nematophila strain. We injected the GFP-labeled bacteria into insects and monitored infection. We found that the bacteria had an extracellular life cycle in the hemolymph and rapidly colonized the anterior midgut region in Spodoptera littoralis larvae. Electron microscopy showed that the bacteria occupied the extracellular matrix of connective tissues within the muscle layers of the Spodoptera midgut. We confirmed the existence of such a specific infection site in the natural route of infection by infesting Spodoptera littoralis larvae with nematodes harboring GFP-labeled Xenorhabdus. When the infective juvenile (IJ) nematodes reached the insect gut, the bacterial cells were rapidly released from the intestinal vesicle into the nematode intestine. Xenorhabdus began to escape from the anus of the nematodes when IJs were wedged in the insect intestinal wall toward the insect hemolymph. Following their release into the insect hemocoel, GFP-labeled bacteria were found only in the anterior midgut region and hemolymph of Spodoptera larvae. Comparative infection assays conducted with another insect, Locusta migratoria, also showed early bacterial colonization of connective tissues. This work shows that the extracellular matrix acts as a particular colonization site for X. nematophila within insects.

Animals↗

The aerodynamics of insect flight.

The flight of insects has fascinated physicists and biologists for more than a century. Yet, until recently, researchers were unable to rigorously quantify the complex wing motions of flapping insects or measure the forces and flows around their wings. However, recent developments in high-speed videography and tools for computational and mechanical modeling have allowed researchers to make rapid progress in advancing our understanding of insect flight. These mechanical and computational fluid dynamic models, combined with modern flow visualization techniques, have revealed that the fluid dynamic phenomena underlying flapping flight are different from those of non-flapping, 2-D wings on which most previous models were based. In particular, even at high angles of attack, a prominent leading edge vortex remains stably attached on the insect wing and does not shed into an unsteady wake, as would be expected from non-flapping 2-D wings. Its presence greatly enhances the forces generated by the wing, thus enabling insects to hover or maneuver. In addition, flight forces are further enhanced by other mechanisms acting during changes in angle of attack, especially at stroke reversal, the mutual interaction of the two wings at dorsal stroke reversal or wing-wake interactions following stroke reversal. This progress has enabled the development of simple analytical and empirical models that allow us to calculate the instantaneous forces on flapping insect wings more accurately than was previously possible. It also promises to foster new and exciting multi-disciplinary collaborations between physicists who seek to explain the phenomenology, biologists who seek to understand its relevance to insect physiology and evolution, and engineers who are inspired to build micro-robotic insects using these principles. This review covers the basic physical principles underlying flapping flight in insects, results of recent experiments concerning the aerodynamics of insect flight, as well as the different approaches used to model these phenomena.

Animals↗

Community structure and its dynamics of pest, predatory and neutral insects in a jujube ecosystem.

An investigation on the insects in the jujube ecosystem in Taigu District of Shanxi Province, Northern China showed that more species and individual numbers of pest, predatory and neutral insects were found on the tree than on the ground. The ratio of the species and individual numbers of predatory and neutral insects to those of pest insects fluctuated from year to year. Homoptera, Coleoptera and Lepidoptera were the dominant groups of pest insects, while those of predatory insects were Coleopteran, Hemiptera, Diptera and Hymenoptera. The vertical distribution of the community structure of the same or different subcommunity was different in different seasons, as was the case of the same or different subcommunity in the same season. The diversity indexes of pest, predatory and neutral insects fluctuated with seasons, and the populations of predatory and neutral insects had a significant correlation (r = 0.9833, P <0.05) with the fluctuation of pest insects. There was also a significant correlation between the pest, predatory and neutral insects in different stratums of tree canopy, especially in the middle stratum of tree canopy (r = 0.9887, P <0.01).

Animals↗

Mutations and their use in insect control.

Traditional chemically based methods for insect control have been shown to have serious limitations, and many alternative approaches have been developed and evaluated, including those based on the use of different types of mutation. The mutagenic action of ionizing radiation was well known in the field of genetics long before it was realized by entomologists that it might be used to induce dominant lethal mutations in insects, which, when released, could sterilize wild female insects. The use of radiation to induce dominant lethal mutations in the sterile insect technique (SIT) is now a major component of many large and successful programs for pest suppression and eradication. Adult insects, and their different developmental stages, differ in their sensitivity to the induction of dominant lethal mutations, and care has to be taken to identify the appropriate dose of radiation that produces the required level of sterility without impairing the overall fitness of the released insect. Sterility can also be introduced into populations through genetic mechanisms, including translocations, hybrid incompatibility, and inherited sterility in Lepidoptera. The latter phenomenon is due to the fact that this group of insects has holokinetic chromosomes. Specific types of mutations can also be used to make improvements to the SIT, especially for the development of strains for the production of only male insects for sterilization and release. These strains utilize male translocations and a variety of selectable mutations, either conditional or visible, so that at some stage of development, the males can be separated from the females. In one major insect pest, Ceratitis capitata, these strains are used routinely in large operational programs. This review summarizes these developments, including the possible future use of transgenic technology in pest control.

Animals↗

Role of phermones and kairmones for insect suppression systems and their possible health and environmental impacts.

Insects produce pheromones as a chemical communication system to facilitate reproduction. These highly active chemical attractants have been synthesized for some of the most important insect pests, including the boll weevil, gypsy moth, codling moth, tobacco budworm, European corn borer, and several bark beetles. While none of the synthetic sex attractants have yet been developed for use in insect control, they offer opportunities for the future both as control agents and to greatly improved insect detection. Investigations are underway on insect trapping systems employing the phermones and on air permeation techniques to disrupt insect reproduction. The pheromones are generally highly species-specific and are not likely to pose hazards to nontarget organisms in the environment. Toxicological studies indicate that they are low in toxicity to mammals, birds, and fish, but adequate toxicological data are necessary before they can be registered for use in insect control. Another new class of compounds called kaironomes has been discovered. These chemicals are involved in the detection of hosts or prey by insect parasites and predators. Kairomones may prove useful in manipulating natural or released biological agents for more effective biological control of insect pests. No information is yet available on the toxicology of these chemicals.

Air↗

Modulation of insect Ca(v) channels by peptidic spider toxins.

Insects have a much smaller repertoire of voltage-gated calcium (Ca(V)) channels than vertebrates. Drosophila melanogaster harbors only a single ortholog of each of the vertebrate Ca(V)1, Ca(V)2, and Ca(V)3 subtypes, although its basal inventory is expanded by alternative splicing and editing of Ca(V) channel transcripts. Nevertheless, there appears to be little functional plasticity within this limited panel of insect Ca(V) channels, since severe loss-of-function mutations in genes encoding the pore-forming alpha1 subunits in Drosophila are embryonic lethal. Since the primary role of spider venom is to paralyze or kill insect prey, it is not surprising that most, if not all, spider venoms contain peptides that potently modify the activity of these functionally critical insect Ca(V) channels. Unfortunately, it has proven difficult to determine the precise ion channel subtypes recognized by these peptide toxins since insect Ca(V) channels have significantly different pharmacology to their vertebrate counterparts, and cloned insect Ca(V) channels are not available for electrophysiological studies. However, biochemical and genetic studies indicate that some of these spider toxins might ultimately become the defining pharmacology for certain subtypes of insect Ca(V) channels. This review focuses on peptidic spider toxins that specifically target insect Ca(V) channels. In addition to providing novel molecular tools for ion channel characterization, some of these toxins are being used as leads to develop new methods for controlling insect pests.

Amino Acid Sequence↗

Molecular characteristics of insect vitellogenins and vitellogenin receptors.

The recent cloning and sequencing of several insect vitellogenins (Vg), the major yolk protein precursor of most oviparous animals, and the mosquito Vg receptor (VgR) has brought the study of insect vitellogenesis to a new plane. Insect Vgs are homologous to nematode and vertebrate Vgs. All but one of the insect Vgs for which we know the primary structure are cleaved into two subunits at a site [(R/K)X(R/K)R or RXXR with an adjacent beta-turn] recognized by subtilisin-like proprotein convertases. In four of the Vgs, the cleavage site is near the N-terminus, but in one insect species, it is near the C-terminus of the Vg precursor. Multiple alignments of these Vg sequences indicate that the variation in cleavage location has not arisen through exon shuffling, but through local modifications of the amino acid sequences. A wasp Vg precursor is not cleaved, apparently because the sequence at the presumed ancestral cleavage site has been mutated from RXRR to LYRR and is no longer recognized by convertases. Some insect Vgs contain polyserine domains which are reminiscent of, but not homologous to, the phosvitin domain in vertebrate Vgs. The sequence of the mosquito VgR revealed that it is a member of the low-density lipoprotein receptor (LDLR) family. Though resembling chicken and frog VgRs, which are also members of the LDLR family, it is twice as big, carrying two clusters of cysteine-rich complement-type (Class A) repeats (implicated in ligand-binding) instead of one like vertebrate VgRs and LDLRs. It is very similar in sequence and domain arrangement to the Drosophila yolk protein receptor (YPR), despite a non-vitellogenin ligand for the latter. Though vertebrate VgRs, insect VgR/YPRs, and LDLR-related proteins/megalins all accommodate one cluster of eight Class A repeats, fingerprint analysis of the repeats in these clusters indicate they are not directly homologous with one another, but have undergone differing histories of duplications, deletions, and exon shuffling so that their apparent similarity is superficial. The so-called epidermal growth factor precursor region contains two types of motifs (cysteine-rich Class B repeats and YWXD repeats) which occur independently of one another in diverse proteins, and are often involved in protein-protein interactions, suggesting that they potentially are involved in dimerization of VgRs and other LDLR-family proteins. Like the LDLR, but unlike vertebrate VgRs and the Drosophila YPR, the mosquito VgR contains a putative O-linked sugar region on the extra-cellular side of the transmembrane domain. Its function is unclear, but may protect the receptor from membrane-bound proteases. The cytoplasmic tail of insect VgR/YPRs contains a di-leucine (or leucine-isoleucine) internalization signal, unlike the tight-turn tyrosine motif of other LDLR-family proteins. The importance of understanding the details of yolk protein uptake by oocytes lies in its potential for exploitation in novel insect control strategies, and the molecular characterization of the proteins involved has made the development of such strategies a realistic possibility.

Amino Acid Sequence↗