Virus maturation is found in all animal viruses and dsDNA bacteriophages that have been studied. It is a programmed process, cued by cellular environmental factors, that transitions a noninfectious, initial assembly product (provirus) to an infectious particle (virion). Nudaurelia capensis omega virus (NωV) is an ssRNA insect virus with T=4 quasi-symmetry. Over the last 20 years, NωV virus-like particles (VLPs) have been an attractive model for the detailed study of maturation. The novel feature of the system is the progressive transition from procapsid to capsid controlled by pH. Homogeneous populations of maturation intermediates can be readily produced at arbitrary intervals by adjusting the pH between 7.6 and 5.0. These intermediates were investigated using biochemical and biophysical methods to create a stop-frame transition series of this complex process. The studies reviewed here characterized the large-scale subunit reorganization during maturation (the particle changes size from 48 nm to 41 nm) as well as the mechanism of a maturation cleavage, a time-resolved study of cleavage site formation, and specific roles of quasi-equivalent subunits in the release of membrane lytic peptides required for cellular entry.

DNA barcoding is used in a variety of ecological applications to identify organisms, including partially digested prey items from diet samples. That particular application can enhance the ability to characterize diet and predator–prey dynamics but is problematic when genetic sequences of prey match those of consumer species (i.e., self-DNA). Such a result may indicate cannibalism, but false positives can result from contamination of degraded prey samples with consumer DNA. Here, nuclear-encoded microsatellite markers were used to genotype invasive lionfish, Pterois volitans, consumers and their prey (n = 80 pairs) previously barcoded as lionfish. Cannibalism was confirmed when samples exhibited two or more different alleles between lionfish and prey DNA across multiple microsatellite loci. This occurred in 26.2% of all samples and in 42% of samples for which the data were considered conclusive. These estimates should be considered conservative given rigorous assignment criteria and low allelic diversity in invasive lionfish populations. The highest incidence of cannibalism corresponded to larger sized consumers from areas with high lionfish densities, suggesting cannibalism in northern Gulf of Mexico lionfish is size- and density-dependent. Cannibalism has the potential to influence population dynamics of lionfish which lack native western Atlantic predators. These results also have important implications for interpreting DNA barcoding analysis of diet in other predatory species where cannibalism may be underreported.

The invasion of the western Atlantic Ocean by the Indo-Pacific red lionfish (Pterois volitans) has had devastating consequences for marine ecosystems. Estimating the number of colonizing lionfish can be useful in identifying the introduction pathway and can inform policy decisions aimed at preventing similar invasions. It is well-established that at least ten lionfish were initially introduced. However, that estimate has not faced probabilistic scrutiny and is based solely on the number of haplotypes in the maternally-inherited mitochondrial control region. To rigorously estimate the number of lionfish that were introduced, we used a forward-time, Wright-Fisher, population genetic model in concert with a demographic, life-history model to simulate the invasion across a range of source population sizes and colonizing population fecundities. Assuming a balanced sex ratio and no Allee effects, the simulations indicate that the Atlantic population was founded by 118 (54–514, 95% HPD) lionfish from the Indo-Pacific, the Caribbean by 84 (22–328, 95% HPD) lionfish from the Atlantic, and the Gulf of Mexico by at least 114 (no upper bound on 95% HPD) lionfish from the Caribbean. Increasing the size, and therefore diversity, of the Indo-Pacific source population and fecundity of the founding population caused the number of colonists to decrease, but with rapidly diminishing returns. When the simulation was parameterized to minimize the number of colonists (high θ and relative fecundity), 96 (48–216, 95% HPD) colonists were most likely. In a more realistic scenario with Allee effects (e.g., 50% reduction in fecundity) plaguing the colonists, the most likely number of lionfish increased to 272 (106–950, 95% HPD). These results, in combination with other published data, support the hypothesis that lionfish were introduced to the Atlantic via the aquarium trade, rather than shipping. When building the model employed here, we made assumptions that minimize the number of colonists, such as the lionfish being introduced in a single event. While we conservatively modelled the introduction pathway as a single release of lionfish in one location, it is more likely that a combination of smaller and larger releases from a variety of aquarium trade stakeholders occurred near Miami, Florida, which could have led to even larger numbers of colonists than simulated here. Efforts to prevent future invasions via the aquarium trade should focus on the education of stakeholders and the prohibition of release, with adequate rewards for compliance and penalties for violations.

Short rotation Eucalyptus plantations offer great potential for increasing wood-fiber production in the southern United States. Eucalyptus plantations can be highly productive (>35 m(3) ha(-1) year but they may use more water than intensively managed pine (primarily Pinus taeda L) plantations. This has raised concern about how expansion of Eucalyptus plantations will affect water resources. We compared tree water use, stem growth, and WUE (kg wood per m(3) water transpired) in adjacent nine-year-old Eucalyptus benthamii and P. taeda plantations with similar stand density and leaf area. Sap flux (F-d, g cm(-2) s(-1)) was measured continuously over one year using thermal dissipation probes. Stem biomass, stem growth, tree water use (E-t, L day(-1)), canopy transpiration per unit leaf area (E-1, mmol m(-2) s(-1)), and canopy stomatal conductance (G(s), mmol m(2) s(-1)) were quantified. Eucalyptus had higher daily Fd (196.6 g cm(-2) day(-1)) and mean daily E-t (24.6 L day(-1)) than pine (105.8 g cm(-2) day(-1), 15.2 L day(-1)). Eucalyptus exhibited a seasonally bimodal pattern in daily E-t that did not occur in pine. Monthly E-t was23-51% higher in Eucalyptus and differences between species were greatest in the spring and fall. Annual E-t was 32% higher in Eucalyptus (9.13 m(3) H2O year(-1)) than pine (5.79 m(3) H2O year(-1)). Annual stem biomass increment was greater in Eucalyptus (Eucalyptus: 22.9; pine: 11.8 kg tree(-1) year(-1)), and Eucalyptus had greater WUE (Eucalyptus: 2.86; pine 1.72 kg biomass m(-3) H2O year(-1)). Pine exhibited a lower seasonal minimum and higher seasonal maximum leaf area index (LAI). At low LAI, there was no significant difference between species in E-l or G(s); however, at maximum LAI, pine E-l and G(s) were 46 and 43%, respectively of rates observed in Eucalyptus. The species differed in G(5) response to vapor pressure deficit (D). At a similar reference G(s) (G(s),(ref) at D =1 kPa), pine exhibited greater stomatal sensitivity to D. These results suggest that (1) Eucalyptus trees had higher sap flux and total water use than pine, (2) Eucalyptus had greater stem growth and WUE, and (3) species differences in water use were driven primarily by differences in E-l and G(s). Published by Elsevier B.V.

The invasion of the western Atlantic Ocean by the Indo-Pacific red lionfish (Pterois volitans) has had devastating consequences for marine ecosystems. Estimating the number of colonizing lionfish can be useful in identifying the introduction pathway and can inform policy decisions aimed at preventing similar invasions. It is well-established that at least ten lionfish were initially introduced. However, that estimate has not faced statistical scrutiny and is based solely on the number of haplotypes in the maternally-inherited mitochondrial control region observed in samples from the western Atlantic Ocean. To rigorously estimate the number of lionfish introduced, we used a forward-time, Wright-Fisher, population genetic model of the mitochondrial control region in concert with a demographic life-history model to simulate the invasion. The colonization of the western North Atlantic and subsequent spread to the Caribbean and Gulf of Mexico were modeled as single introductions of 1-800 females, which should conservatively estimate the most likely number of colonizing lionfish relative to a model with multiple introductions. Assuming a balanced sex ratio and no Allee effects, the simulations indicate that the Atlantic population was most likely founded by 118 (54-514, 95% HPD) lionfish from the Indo-Pacific, the Caribbean by 84 (22-328, 95% HPD) lionfish from the Atlantic, and the Gulf of Mexico by at least 114 (no upper bound on 95% HPD) lionfish from the Caribbean. Considering the additional possibilities that (1) multiple Indo-Pacific populations colonized the Atlantic and (2) Allee effects (e.g. 50% reduction in fecundity) plagued the colonists, the most likely number of lionfish increased to 180 (88-416, 95% HPD). These results, in combination with all other published data, support the hypothesis that lionfish were introduced to the Atlantic via the aquarium trade, rather than shipping. The most parsimonious introduction pathway is a single release of lionfish in one location, but it seems likely that a combination of smaller and larger releases from a variety of aquarium trade stakeholders occurred near Miami, Florida. Efforts to prevent future invasions via the aquarium trade should focus on the education of stakeholders and the prohibition of release, with adequate rewards for compliance and penalties for violations.

Indo-Pacific lionfish (Pterois volitans and P. miles) recently invaded Western Atlantic waters, rapidly spreading through the Caribbean and Gulf of Mexico (GoM). Previous genetic analyses using the mitochondrial d-loop determined that populations in the Western North Atlantic (NA) region have up to nine haplotypes, whereas Caribbean populations contain four of the North Atlantic haplotypes. The genetic composition of GoM populations, reported here for the first time, could lend insight into the pathway of dispersal into the GoM and better understanding of the biogeography of this recent invader. Here, we determined the genetic composition of lionfish throughout the GoM and compared haplotype composition to Caribbean and North Atlantic regions. We found that GoM samples contained only three d-loop haplotypes that are common in the Caribbean and North Atlantic. The genetic structure differed significantly among the three regions (AMOVA:Φ CT = 0.062; p = 0.001), but we found no differences between locations within regions (AMOVA:Φ SC = 0.005; p = 0.092). The composition of GoM samples most closely matches the composition of Caribbean samples indicating that Caribbean populations are the likely source of the GoM populations. As each region was successively invaded, a drop in haplotype diversity and changes in haplotype frequencies occurred indicating dispersal limitation across basin boundaries and founder effects within each basin. The lack of differentiation within regions indicates rapid population growth and unfettered dispersal within basins after initial colonization. We find no evidence of secondary invasions within samples. With well-established populations, the probability of detecting a secondary invasion is minuscule.

Cystoviruses are dsRNA viruses that infect bacteria and include bacteriophages Φ8 and Φ6. In this issue of Structure, El Omari and colleagues and Nemecek and colleagues report crystal structures of capsid protein P1 pentamers found in procapsid cores of Φ8 and Φ6. The two structures show a striking resemblance in the absence of sequence similarity and offer new mechanistic and evolutionary insights.

Setting records: Native mass spectometry was used to study the assembly of the 18 MDa capsid of bacteriophage HK97. The previous record for the analysis of capsids by this method was around 10 MDa. The results indicate that the efficiency of desolvation is the main priority in improving native MS instrumentation. As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

The capsids of tailed-DNA bacteriophages first assemble as procapsids, which mature by converting into a new form that is strong enough to contain a densely packed viral chromosome. We demonstrate that the intersubunit crosslinking that occurs during maturation of HK97 capsids actually promotes the structural transformation. Small-angle X-ray scattering and crosslinking assays reveal that a shift in the crosslink pattern accompanies conversion of a semimature particle, Expansion Intermediate-I/II, to a more mature state, Balloon. This transition occurs in a switch-like fashion. We find that crosslink formation shifts the global conformational balance to favor the balloon state. A pseudoatomic model of EI-I/II derived from cryo-EM provides insight into the relationship between crosslink formation and conformational switching.

Protein subunits of several RNA viruses are known to undergo post-assembly, autocatalytic cleavage that is required for infectivity. Nudaurelia capensis omega virus (Nomega V) is one of the simplest viruses to undergo an autocatalytic cleavage, making it an excellent model to understand both assembly and the mechanism of autoproteolysis. Heterologous expression of the coat protein gene of Nomega V in a baculovirus system results in the spontaneous assembly of virus-like particles (VLPs) that remain uncleaved when purified at neutral pH. After acidification to pH 5.0, the VLPs autocatalytically cleave at residue 570, providing an in vitro control of the cleavage. The crystal structure of Nomega V displays three residues near the scissile bond that were candidates for participation in the reaction. These were changed by site-directed mutagenesis to conservative and nonconservative residues and the products analyzed. Even conservative changes at the three residues dramatically reduced cleavage when the subunits assembled properly. Unexpectedly, we discovered that these residues are not only critical to the kinetics of Nomega V autoproteolysis, but are also necessary for proper folding of subunits and, ultimately, assembly of Nomega V VLPs.

Large-scale reorganization of protein interactions characterizes many biological processes, yet few systems are accessible to biophysical studies that display this property. The capsid protein of Nudaurelia capensis omega Virus (NomegaV) has previously been characterized in two dramatically different T = 4 quasi-equivalent assembly states when expressed as virus-like particles (VLPs) in a baculovirus system. The procapsid (pH 7), is round, porous, and approximately 450 A in diameter. It converts, in vitro, to the capsid form at pH 5 and the capsid is sealed shut, shaped like an icosahedron, has a maximum diameter of 410 A and undergoes an autocatalytic cleavage at residue 570. Residues 571-644, the gamma peptide, remain associated with the particle and are partially ordered. The interconversion of these states has been previously studied by solution X-ray scattering, electron cryo microscopy (CryoEM), and site-directed mutagenesis. The particle structures appear equivalent in authentic virions and the low pH form of the expressed and assembled protein. Previously, and before the discovery of the multiple morphological forms of the VLPs, we reported the X-ray structure of authentic NomegaV at 2.8 A resolution. These coordinates defined the fold of the protein but were not refined at the time because of technical issues associated with the approximately 2.5 million reflection data set. We now report the refined, authentic virus structure that has added 29 residues to the original model and allows the description of the chemistry of molecular switching for T = 4 capsid formation and the multiple morphological forms. The amino and carboxy termini are internal, predominantly helical, and disordered to different degrees in the four structurally independent subunits; however, the refined structure shows significantly more ordered residues in this region, particularly at the amino end of the B subunit that is now seen to invade space occupied by the A subunits. These additional residues revealed a previously unnoticed strong interaction between the pentameric, gamma peptide helices of the A and B subunits that are largely proximal to the quasi-6-fold axes. One C-terminal helix is ordered in the C and D subunits and stabilizes a flat interaction in two interfaces between the protein monomers while the other, quasi-equivalent, interactions are bent. As this helix is arginine rich, the comparable, disordered region in the A and B subunits probably interacts with RNA. One of the subunit-subunit interfaces has an unusual arrangement of carboxylate side chains. Based on this observation, we propose a mechanism for the control of the pH-dependent transitions of the virus particle.

Picorna-like plant viruses that include Como and Nepo genera of the Comoviridae, share similarities in structure, genome organization, and replication strategy with mammalian picornaviruses. Structural studies of comoviruses and a nepovirus contribute to the understanding of icosahedral virus structures, the development of novel methods for investigating macromolecular assemblies, and the infrastructure for virus-based bio- and nanotechnology. It is found that bean pod mottle virus (BPMV) shares similar overall genome organization, replication strategy, and capsid structure with the type member cowpea mosaic virus (CPMV). Structural studies of BPMV provided the first observation of a portion of a viral genome in association with its capsid, and the structural comparison of a nucleoprotein particle with an empty capsid that demonstrated ordering of elements of the capsid. Red clover mottle virus (RCMV) is another member of the comovirus family that was investigated biochemically and genetically and its structure was determined for comparative studies. There are significant structural differences between comoviruses and some nepoviruses in that the capsids of comoviruses are composed of two polypeptides whereas those of nepoviruses are composed of a single polypeptide. The structure of a nepovirus, tobacco ringspot virus (TRSV), was determined. Structural comparison showed that TRSV was likely to be an evolutionary intermediate of development of comoviruses.

ABSTRACT The assembly and maturation of the coat protein of a T=4, nonenveloped, single-stranded RNA virus, Nudaurelia capensis ω virus (NωV), was examined by using a recombinant baculovirus expression system. At pH 7.6, the coat protein assembles into a stable particle called the procapsid, which is 450 Å in diameter and porous. Lowering the pH to 5.0 leads to a concerted reorganization of the subunits into a 410-Å-diameter particle called the capsid, which has no obvious pores. This conformational change is rapid but reversible until slow, autoproteolytic cleavage occurs in at least 15% of the subunits at the lower pH. In this report, we show that expression of subunits with replacement of Asn-570, which is at the cleavage site, with Thr results in assembly of particles with expected morphology but that are cleavage defective. The conformational change from procapsid to capsid is reversible in N570T mutant virus-like particles, in contrast to wild-type particles, which are locked into the capsid conformation after cleavage of the coat protein. The reexpanded procapsids display slightly different properties than the original procapsid, suggesting hysteretic effects. Because of the stability of the procapsid under near-neutral conditions and the reversible properties of the cleavage-defective mutant, NωV provides an excellent model for the study of pH-induced conformational changes in macromolecular assemblies. Here, we identify the relationship between cleavage and the conformational change and propose a pH-dependent helix-coil transition that may be responsible for the structural rearrangement in NωV.

Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Twitter Facebook Reddit LinkedIn Tools Icon Tools Reprints and Permissions Cite Icon Cite Search Site Citation John C. Johnson; Snowdon, John C. ⋅ 1932–1980. J. Acoust. Soc. Am. 1 September 1980; 68 (3): 993. https://doi.org/10.1121/1.384791 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAcoustical Society of AmericaThe Journal of the Acoustical Society of America Search Advanced Search |Citation Search

The anticoccidial agent salinomycin (AHR-3096) was compared at various treatment levels (60 to 100 p.p.m.) in two experiments against unmedicated and either 100 or 121 p.p.m. monensin-medicated groups to evaluate its performance under floor-pen conditions. Various methods for the initial rearing of birds and coccidial exposure were employed to test the efficacy of salinomycin against severe, moderate, and mild coccidiosis. Salinomycin at all treatment levels showed definite anticoccidial activity resulting in improved weights, feed conversion ratios, lesion scores, and mortality compared with unmedicated controls. In pens with severe coccidiosis, this compound demonstrated a significant improvement in all parameters when compared with the corresponding unmedicated controls. Salinomycin-treated pens exposed to light coccidiosis usually had a numerical increase in the parameters over the unmedicated control groups. Statistical analysis of overall main effects on mortality, weight gain, and feed conversion ratio showed no significant differences between the activity of any level of salinomycin and monensin.