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1.
Ecol Lett ; 21(4): 506-515, 2018 04.
Article in English | MEDLINE | ID: mdl-29446220

ABSTRACT

Species coexistence in diverse communities likely results from multiple interacting factors. Mechanisms such as conspecific negative density dependence (CNDD) and varying life-history strategies related to resource partitioning are known to influence plant fitness, and thereby community composition and diversity. However, we have little understanding of how these mechanisms interact and how they vary across life stages. Here, we document the interaction between CNDD and life-history strategy, based on growth-mortality trade-offs, from seedling to adult tree for 47 species in a tropical forest. Species' life-history strategies remained consistent across stages: fast-growing species had higher mortality than slow-growing species at all stages. In contrast, mean CNDD was strongest at early life stages (i.e. seedling, sapling). Fast-growing species tended to suffer greater CNDD than slow-growing species at several, but not all life stages. Overall, our results demonstrate that coexistence mechanisms interact across multiple life stages to shape diverse tree communities.


Subject(s)
Forests , Trees , Life History Traits , Seedlings , Tropical Climate
2.
Nature ; 507(7490): 90-3, 2014 Mar 06.
Article in English | MEDLINE | ID: mdl-24429523

ABSTRACT

Forests are major components of the global carbon cycle, providing substantial feedback to atmospheric greenhouse gas concentrations. Our ability to understand and predict changes in the forest carbon cycle--particularly net primary productivity and carbon storage--increasingly relies on models that represent biological processes across several scales of biological organization, from tree leaves to forest stands. Yet, despite advances in our understanding of productivity at the scales of leaves and stands, no consensus exists about the nature of productivity at the scale of the individual tree, in part because we lack a broad empirical assessment of whether rates of absolute tree mass growth (and thus carbon accumulation) decrease, remain constant, or increase as trees increase in size and age. Here we present a global analysis of 403 tropical and temperate tree species, showing that for most species mass growth rate increases continuously with tree size. Thus, large, old trees do not act simply as senescent carbon reservoirs but actively fix large amounts of carbon compared to smaller trees; at the extreme, a single big tree can add the same amount of carbon to the forest within a year as is contained in an entire mid-sized tree. The apparent paradoxes of individual tree growth increasing with tree size despite declining leaf-level and stand-level productivity can be explained, respectively, by increases in a tree's total leaf area that outpace declines in productivity per unit of leaf area and, among other factors, age-related reductions in population density. Our results resolve conflicting assumptions about the nature of tree growth, inform efforts to undertand and model forest carbon dynamics, and have additional implications for theories of resource allocation and plant senescence.


Subject(s)
Body Size , Carbon Cycle , Carbon/metabolism , Trees/anatomy & histology , Trees/metabolism , Aging/metabolism , Biomass , Climate , Geography , Models, Biological , Plant Leaves/growth & development , Plant Leaves/metabolism , Sample Size , Species Specificity , Time Factors , Trees/classification , Trees/growth & development , Tropical Climate
3.
Ecology ; 89(7): 1908-20, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18705377

ABSTRACT

A central goal of comparative plant ecology is to understand how functional traits vary among species and to what extent this variation has adaptive value. Here we evaluate relationships between four functional traits (seed volume, specific leaf area, wood density, and adult stature) and two demographic attributes (diameter growth and tree mortality) for large trees of 240 tree species from five Neotropical forests. We evaluate how these key functional traits are related to survival and growth and whether similar relationships between traits and demography hold across different tropical forests. There was a tendency for a trade-off between growth and survival across rain forest tree species. Wood density, seed volume, and adult stature were significant predictors of growth and/or mortality. Both growth and mortality rates declined with an increase in wood density. This is consistent with greater construction costs and greater resistance to stem damage for denser wood. Growth and mortality rates also declined as seed volume increased. This is consistent with an adaptive syndrome in which species tolerant of low resource availability (in this case shade-tolerant species) have large seeds to establish successfully and low inherent growth and mortality rates. Growth increased and mortality decreased with an increase in adult stature, because taller species have a greater access to light and longer life spans. Specific leaf area was, surprisingly, only modestly informative for the performance of large trees and had ambiguous relationships with growth and survival. Single traits accounted for 9-55% of the interspecific variation in growth and mortality rates at individual sites. Significant correlations with demographic rates tended to be similar across forests and for phylogenetically independent contrasts as well as for cross-species analyses that treated each species as an independent observation. In combination, the morphological traits explained 41% of the variation in growth rate and 54% of the variation in mortality rate, with wood density being the best predictor of growth and mortality. Relationships between functional traits and demographic rates were statistically similar across a wide range of Neotropical forests. The consistency of these results strongly suggests that tropical rain forest species face similar trade-offs in different sites and converge on similar sets of solutions.


Subject(s)
Ecosystem , Plant Development , Trees/physiology , Tropical Climate , Plant Leaves/growth & development , Population Dynamics , Seedlings
4.
Science ; 293(5530): 650-5, 2001 Jul 27.
Article in English | MEDLINE | ID: mdl-11474101

ABSTRACT

By integrating a wide range of experimental, comparative, and theoretical approaches, ecologists are starting to gain a detailed understanding of the long-term dynamics of vegetation. We explore how patterns of variation in demographic traits among species have provided insight into the processes that structure plant communities. We find a common set of mechanisms, derived from ecological and evolutionary principles, that underlie the main forces shaping systems as diverse as annual plant communities and tropical forests. Trait variation between species maintains diversity and has important implications for ecosystem processes. Hence, greater understanding of how Earth's vegetation functions will likely require integration of ecosystem science with ideas from plant evolutionary, population, and community ecology.


Subject(s)
Ecosystem , Plants , Animals , Biological Evolution , Climate , Environment , Plant Development , Poaceae/growth & development , Population Density , Population Dynamics , Seeds , Time Factors , Trees , Tropical Climate
5.
Virology ; 280(1): 143-52, 2001 Feb 01.
Article in English | MEDLINE | ID: mdl-11162828

ABSTRACT

J3R, the 39-kDa subunit of vaccinia virus poly(A) polymerase, is a multifunctional protein that catalyzes (nucleoside-2'-O-)-methyltransferase activity, serves as a poly(A) polymerase stimulatory factor, and acts as a postreplicative positive transcription elongation factor. Prior results support an association between poly(A) polymerase and the virion RNA polymerase. A possible direct interaction between J3R and H4L subunit of virion RNA polymerase was evaluated. J3R was shown to specifically bind to H4L amino acids 235-256, C terminal to NPH I binding site on H4L. H4L binds to the C-terminal region of J3R between amino acids 169 and 333. The presence of a J3R binding site near to the NPH I binding region on H4L led us to evaluate a physical interaction between NPH I and J3R. The NPH I binding site was located on J3R between amino acids 169 and 249, and J3R was shown to bind to NPH I between amino acids 457 and 524. To evaluate a role for J3R in early gene mRNA synthesis, transcription termination, and/or release, a transcription-competent extract prepared from cells infected with mutant virus lacking J3R, J3-7. Analysis of transcription activity demonstrated that J3R is not required for early mRNA synthesis and is not an essential factor in early gene transcription termination or transcript release in vitro. J3R interaction with NPH I and H4L may serve as a docking site for J3R on the virion RNA polymerase, linking transcription to mRNA cap formation and poly(A) addition.


Subject(s)
DNA-Directed RNA Polymerases/metabolism , Polynucleotide Adenylyltransferase/metabolism , Transcription Factors/metabolism , Vaccinia virus/enzymology , Viral Proteins/metabolism , Animals , Cell Line , Chlorocebus aethiops
6.
Virology ; 277(2): 439-49, 2000 Nov 25.
Article in English | MEDLINE | ID: mdl-11080491

ABSTRACT

The biological properties of poxvirus isolates from skin lesions on dairy cows and milkers during recent exanthem episodes in Cantagalo County, Rio de Janeiro State, Brazil, were more like vaccinia virus (VV) than cowpox virus. PCR amplification of the hemagglutinin (HA) gene substantiated the isolate classification as an Old World orthopoxvirus, and alignment of the HA sequences with those of other orthopoxviruses indicated that all the isolates represented a single strain of VV, which we have designated Cantagalo virus (CTGV). HA sequences of the Brazilian smallpox vaccine strain (VV-IOC), used over 20 years ago, and CTGV showed 98.2% identity; phylogeny inference of CTGV, VV-IOC, and 12 VV strains placed VV-IOC and CTGV together in a distinct clade. Viral DNA restriction patterns and protein profiles showed a few differences between VV-IOC and CTGV. Together, the data suggested that CTGV may have derived from VV-IOC by persisting in an indigenous animal(s), accumulating polymorphisms, and now emerging in cattle and milkers as CTGV. CTGV may represent the first case of long-term persistence of vaccinia in the New World.


Subject(s)
Cattle Diseases/virology , Disease Outbreaks/veterinary , Poxviridae Infections/veterinary , Poxviridae/classification , Smallpox Vaccine , Amino Acid Sequence , Animals , Brazil/epidemiology , Cattle , Cattle Diseases/epidemiology , Chlorocebus aethiops , Exanthema/epidemiology , Exanthema/virology , Female , Hemagglutinins, Viral/genetics , Humans , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , Poxviridae/genetics , Poxviridae/isolation & purification , Poxviridae Infections/virology , Sequence Alignment , Vaccinia virus/genetics , Vaccinia virus/immunology , Vero Cells
7.
Proc Natl Acad Sci U S A ; 97(20): 10850-4, 2000 Sep 26.
Article in English | MEDLINE | ID: mdl-11005859

ABSTRACT

A fundamental question in ecology is how many species occur within a given area. Despite the complexity and diversity of different ecosystems, there exists a surprisingly simple, approximate answer: the number of species is proportional to the size of the area raised to some exponent. The exponent often turns out to be roughly 1/4. This power law can be derived from assumptions about the relative abundances of species or from notions of self-similarity. Here we analyze the largest existing data set of location-mapped species: over one million, individually identified trees from five tropical forests on three continents. Although the power law is a reasonable, zeroth-order approximation of our data, we find consistent deviations from it on all spatial scales. Furthermore, tropical forests are not self-similar at areas

8.
Science ; 288(5470): 1414-8, 2000 May 26.
Article in English | MEDLINE | ID: mdl-10827950

ABSTRACT

Fully mapped tree census plots of large area, 25 to 52 hectares, have now been completed at six different sites in tropical forests, including dry deciduous to wet evergreen forest on two continents. One of the main goals of these plots has been to evaluate spatial patterns in tropical tree populations. Here the degree of aggregation in the distribution of 1768 tree species is examined based on the average density of conspecific trees in circular neighborhoods around each tree. When all individuals larger than 1 centimeter in stem diameter were included, nearly every species was more aggregated than a random distribution. Considering only larger trees (>/= 10 centimeters in diameter), the pattern persisted, with most species being more aggregated than random. Rare species were more aggregated than common species. All six forests were very similar in all the particulars of these results.


Subject(s)
Ecosystem , Trees , Central America , India , Malaysia , Panama , Sri Lanka , Statistics as Topic , Thailand , Tropical Climate
9.
J Biol Chem ; 275(2): 1485-94, 2000 Jan 14.
Article in English | MEDLINE | ID: mdl-10625702

ABSTRACT

Prior phenotypic analysis of a vaccinia virus gene A18R mutant, Cts23, showed the synthesis of longer than wild type (Wt) length viral transcripts during the intermediate stage of infection, indicating that the A18R protein may act as a negative transcription elongation factor. The purpose of the work described here was to determine a biochemical activity for the A18R protein. Pulse-labeled transcription complexes established from intermediate virus promoters on bead-bound DNA templates were assayed for transcript release during an elongation step that contained nucleotides and various proteins. Pulse-labeled transcription complexes elongated in the presence of only nucleotides were unable to release nascent RNA. The addition of Wt extract during the elongation phase resulted in release of the nascent transcript, indicating that additional factors present in the Wt extract are capable of inducing transcript release. Extract from Cts23 or mock-infected cells was unable to induce release. The lack of release upon addition of Cts23 extract suggests that A18R is involved in release of nascent RNA. By itself, purified polyhistidine-tagged A18R protein (His-A18R) was unable to induce release; however, release did occur in the presence of purified His-A18R protein plus extract from either Cts23 or mock-infected cells. These data taken together indicate that A18R is necessary but not sufficient for release of nascent transcripts. We have also demonstrated that the combination of A18R protein and mock extract induces transcript release in an ATP-dependent manner, consistent with the fact that the A18R protein is an ATP-dependent helicase. Further analysis revealed that the release activity is not restricted to a vaccinia intermediate promoter but is observed using pulse-labeled transcription complexes initiated from all three viral gene class promoters. Therefore, we conclude that A18R and an as yet unidentified cellular factor(s) are required for the in vitro release of nascent RNA from a vaccinia virus transcription elongation complex.


Subject(s)
Adenosine Triphosphatases/metabolism , DNA Helicases/metabolism , Transcription, Genetic , Vaccinia virus/enzymology , Vaccinia virus/genetics , Adenosine Triphosphatases/genetics , Adenosine Triphosphatases/isolation & purification , Chromatography, Ion Exchange , Cloning, Molecular , Escherichia coli , Humans , Kinetics , Phenotype , Promoter Regions, Genetic , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Templates, Genetic , Tumor Cells, Cultured
10.
Virology ; 269(2): 345-55, 2000 Apr 10.
Article in English | MEDLINE | ID: mdl-10753713

ABSTRACT

Vaccinia virus genes A18 and G2 affect the elongation and termination of postreplicative viral gene transcription in opposite ways. Viruses with mutations in gene A18 produce abnormally long transcripts, indicating that A18 is a negative transcription elongation factor. Viruses containing mutations in gene G2 produce transcripts that are abnormally short, truncated specifically from their 3' ends, indicating that G2 is a positive transcription elongation factor. Despite the fact that both A18 and G2 are essential genes, A18-G2 double-mutant viruses are viable, presumably because the effects of the mutations are mutually compensatory. In addition, the anti-poxviral drug isatin-beta-thiosemicarbazone (IBT) seems to enhance elongation during a vaccinia infection: IBT treatment of a wildtype vaccinia infection induces a phenotype identical to an A18 mutant infection, and G2 mutant viruses are dependent on IBT for growth, presumably because IBT restores the G2 mutant truncated transcripts to a normal length. These observations inspire two independent genetic selections that have now been used to identify an additional vaccinia gene, J3, that regulates postreplicative transcription elongation. In the first selection, a single virus that contains an extragenic suppressor of the A18 temperature-sensitive mutant, Cts23, was isolated. In the second selection, several spontaneous IBT-dependent (IBT(d)) mutant viruses were isolated and characterized genetically. Marker rescue mapping and DNA sequence analysis show that the extragenic suppressor of Cts23 contains a point mutation in the J3 gene, while each of seven new IBT(d) mutants contains null mutations in the J3 gene. The J3 protein has previously been identified as a (nucleoside-2'-O-)-methyltransferase and as a processivity subunit for the heterodimeric viral poly(A) polymerase. The nature of the two independent selections used to isolate the J3 mutants strongly suggests that the J3 protein serves as a positive postreplicative transcription elongation factor during a normal virus infection.


Subject(s)
Genes, Viral , Peptide Elongation Factors/genetics , Polynucleotide Adenylyltransferase/metabolism , Suppression, Genetic , Vaccinia virus/genetics , Animals , Blotting, Western , Chlorocebus aethiops , Genetic Markers , Mutagenesis , Point Mutation , Sequence Analysis, DNA , Vero Cells
11.
Virology ; 269(2): 356-69, 2000 Apr 10.
Article in English | MEDLINE | ID: mdl-10753714

ABSTRACT

Prior genetic analysis suggests that the vaccinia virus J3 gene product, previously characterized as a bifunctional (nucleoside-2'-O-)-methyltransferase and poly(A) polymerase stimulatory factor, is a postreplicative positive transcription elongation factor. To test this hypothesis, viruses bearing mutations in the J3 gene were characterized with respect to viral protein and RNA synthesis in infected cells. The analysis reveals that compared to wt virus infections, J3 mutants synthesize reduced amounts of large late viral proteins and shorter-than-normal intermediate and late mRNAs. Structural analysis of one late mRNA shows that it is specifically truncated from the 3' end, thus accounting for its shorter than normal chain length. Thus J3 mutant viruses are defective in elongation of transcription of postreplicative viral genes, strongly suggesting that the J3 gene product normally acts as a positive transcription elongation factor. Biochemical analysis of one J3 missense mutant demonstrates that it retains poly(A) stimulatory activity but is defective in (nucleoside-2'-O-)-methyltransferase activity. Thus the elongation factor activity of the J3 gene product is independent of the poly(A) stimulatory activity. It remains to be determined whether the (nucleoside-2'-O-)-methyltransferase and elongation factor activities of the J3 protein are linked or can be uncoupled by mutation.


Subject(s)
Polynucleotide Adenylyltransferase/metabolism , Transcription Factors/physiology , Transcription, Genetic , Vaccinia virus/metabolism , Animals , Chlorocebus aethiops , Electrophoresis, Polyacrylamide Gel , Histidine/metabolism , Mutagenesis, Site-Directed , Poly A/metabolism , Transcription Factors/genetics , Vaccinia virus/genetics , Vero Cells
12.
Proc Biol Sci ; 266(1427): 1445-52, 1999 Jul 22.
Article in English | MEDLINE | ID: mdl-10457617

ABSTRACT

Quadrat-based analysis of two rainforest plots of area 50 ha, one in Panama (Barro Colorado Island, BCI) and the other in Malaysia (Pasoh), shows that in both plots recruitment is in general negatively correlated with both numbers and biomass of adult trees of the same species in the same quadrat. At BCI, this effect is not significantly influenced by treefall gaps. In both plots, recruitment of individual species is negatively correlated with the numbers of trees of all species in the quadrats, but not with overall biomass. These observations suggest, but do not prove, widespread frequency-dependent effects produced by pathogens and seed-predators that act most effectively in quadrats crowded with trees. Within-species correlations of mortality with numbers or biomass are not found in either plot, indicating that most frequency-dependent mortality takes place before the trees reach 1 cm in diameter. Stochastic effects caused by BCI's more rapid tree turnover may contribute to a larger variance in diversity from quadrat to quadrat at BCI, although they are not sufficient to explain why BCI has fewer than half as many tree species as Pasoh. Finally, in both plots quadrats with low diversity show a significant increase in diversity over time, and this increase is stronger at BCI. This process, like the frequency-dependence, will tend to maintain diversity over time. In general, these non-random forces that should lead to the maintenance of diversity are slightly stronger at BCI, even though the BCI plot is less diverse than the Pasoh plot.


Subject(s)
Ecosystem , Trees/physiology , Forestry/methods , Stochastic Processes , Tropical Climate
13.
Science ; 283(5401): 554-7, 1999 Jan 22.
Article in English | MEDLINE | ID: mdl-9915706

ABSTRACT

Light gap disturbances have been postulated to play a major role in maintaining tree diversity in species-rich tropical forests. This hypothesis was tested in more than 1200 gaps in a tropical forest in Panama over a 13-year period. Gaps increased seedling establishment and sapling densities, but this effect was nonspecific and broad-spectrum, and species richness per stem was identical in gaps and in nongap control sites. Spatial and temporal variation in the gap disturbance regime did not explain variation in species richness. The species composition of gaps was unpredictable even for pioneer tree species. Strong recruitment limitation appears to decouple the gap disturbance regime from control of tree diversity in this tropical forest.

14.
Philos Trans R Soc Lond B Biol Sci ; 354(1391): 1739-48, 1999 Nov 29.
Article in English | MEDLINE | ID: mdl-11605618

ABSTRACT

Dynamics of the Pasoh forest in Peninsular Malaysia were assessed by drawing a comparison with a forest in Panama, Central America, whose dynamics have been thoroughly described. Census plots of 50 ha were established at both sites using standard methods. Tree mortality at Pasoh over an eight-year interval was 1.46% yr(-1) for all stems > or = 10 mm diameter at breast height (dbh), and 1.48% yr(-1) for stems > or = 100 mm dbh. Comparable figures at the Barro Colorado Island site in Panama (BCI) were 2.55% and 2.03%. Growth and recruitment rates were likewise considerably higher at BCI than at Pasoh. For example, in all trees 500-700 mm in dbh, mean BCI growth over the period 1985-1995 was 6 mm yr(-1), whereas mean Pasoh growth was about 3.5 mm yr(-1). Examining growth and mortality rates for individual species showed that the difference between the forests can be attributed to a few light-demanding pioneer species at BCI, which have very high growth and mortality; Pasoh is essentially lacking this guild. The bulk of the species in the two forests are shade-tolerant and have very similar mortality, growth and recruitment. The Pasoh forest is more stable than BCI's in another way as well: few of its tree populations changed much over the eight-year census interval. In contrast, at BCI, over 10% of the species had populations increasing or decreasing at a rate of >0.05 yr(-1) compared to just 2% of the species at Pasoh). The faster species turnover at BCI can probably be attributed to severe droughts that have plagued the forest periodically over the past 30 years; Pasoh has not suffered such extreme events recently. The dearth of pioneer species at Pasoh is associated with low-nutrient soil and slow litter breakdown, but the exact mechanisms behind this association remain poorly understood.


Subject(s)
Trees , Ecosystem , Malaysia , Panama , Trees/growth & development , Tropical Climate
15.
Proc Natl Acad Sci U S A ; 95(23): 13917-22, 1998 Nov 10.
Article in English | MEDLINE | ID: mdl-9811901

ABSTRACT

Vaccinia uses actin-based motility for virion movement in host cells, but the specific protein components have yet to be defined. A cardinal feature of Listeria and Shigella actin-based motility is the involvement of vasodilator-stimulated phosphoprotein (VASP). This essential adapter recognizes and binds to actin-based motility 1 (ABM-1) consensus sequences [(D/E)FPPPPX(D/E), X = P or T] contained in Listeria ActA and in the p90 host-cell vinculin fragment generated by Shigella infection. VASP, in turn, provides the ABM-2 sequences [XPPPPP, X = G, P, L, S, A] for binding profilin, an actin-regulatory protein that stimulates actin filament assembly. Immunolocalization using rabbit anti-VASP antibody revealed that VASP concentrates behind motile virions in HeLa cells. Profilin was also present in these actin-rich rocket tails, and microinjection of 10 microM (intracellular) ABM-2 peptide (GPPPPP)3 blocked vaccinia actin-based motility. Vinculin did not colocalize with VASP on motile virions and remained in focal adhesion contacts; however, another ABM-1-containing host protein, zyxin, was concentrated at the rear of motile virions. We also examined time-dependent changes in the location of these cytoskeletal proteins during vaccinia infection. VASP and zyxin were redistributed dramatically several hours before the formation of actin rocket tails, concentrating in the viral factories of the perinuclear cytoplasm. Our findings underscore the universal involvement of ABM-1 and ABM-2 docking sites in actin-based motility of Listeria, Shigella, and now vaccinia.


Subject(s)
Actins/physiology , Vaccinia virus/physiology , Animals , Biological Transport, Active , Cytoskeleton/virology , HeLa Cells , Humans , Peptide Fragments , Rabbits , Virus Replication
16.
J Virol ; 72(9): 7012-23, 1998 Sep.
Article in English | MEDLINE | ID: mdl-9696793

ABSTRACT

Loss of vaccinia virus A18R gene function results in an aberrant transcription profile termed promiscuous transcription, defined as transcription within regions of the genome which are normally transcriptionally silent late during infection. Promiscuous transcription results in an increase in the intracellular concentration of double-stranded RNA, which in turn results in activation of the cellular 2-5A pathway and subsequent RNase L-catalyzed degradation of viral and cellular RNAs. One of three hypotheses could account for promiscuous transcription: (i) reactivation of early promoters late during infection, (ii) random transcription initiation, (iii) readthrough transcription from upstream promoters. Transcriptional analysis of several viral genes, presented here, argues strongly against the first two hypotheses. We have tested the readthrough hypothesis by conducting a detailed transcriptional analysis of a region of the vaccinia virus genome which contains three early genes (M1L, M2L, and K1L) positioned directly downstream of the intermediate gene, K2L. The results show that mutation of the A18R gene results in increased readthrough transcription of the M1L gene originating from the K2L intermediate promoter. A18R mutant infection of RNase L knockout mouse fibroblast (KO3) cells does not result in 2-5A pathway activation, yet the virus mutant is defective in late viral gene expression and remains temperature sensitive. These results demonstrate that the A18R gene product is a negative transcription elongation factor for postreplicative viral genes.


Subject(s)
Adenosine Triphosphatases/genetics , DNA Helicases/genetics , DNA Replication , Gene Expression Regulation, Viral , Genes, Viral , Peptide Elongation Factors/genetics , Vaccinia virus/genetics , Virus Replication , Animals , Cell Line , Chlorocebus aethiops , Endoribonucleases/genetics , Mice , Mice, Knockout , Mutagenesis , Phenotype , Polymerase Chain Reaction/methods , Vaccinia virus/physiology
17.
Virology ; 245(2): 313-22, 1998 Jun 05.
Article in English | MEDLINE | ID: mdl-9636370

ABSTRACT

Prior genetic analysis suggests that there may exist an interaction between the products of the vaccinia virus genes A18R, a putative negative transcription elongation factor, and G2R, a putative positive transcription elongation factor. In addition, affinity purification of polyhistidine-tagged G2R protein overexpressed in vaccinia virus-infected cells, reported here, results in copurification of the vaccinia H5R protein, previously characterized as a late viral transcription factor. We have therefore used several methods to screen further for interactions among the G2R, A18R, and H5R proteins. Methods include copurification or co-immunoprecipitation of proteins overexpressed during vaccinia virus infection, activation of the gal 4 promoter by gal 4 fusions in the yeast two-hybrid system, and co-immunoprecipitation of proteins synthesized in vitro in a rabbit reticulocyte lysate. The results reveal interactions which include all possible pairwise combinations of the three proteins G2R, A18R, and H5R; however, not all possible permutations of the interactions are observed and the interactions are not observed in all environments tested. The results suggest that the vaccinia virus proteins G2R, A18R, and H5R interact as part of a higher order transcription complex.


Subject(s)
Adenosine Triphosphatases/genetics , Adenosine Triphosphatases/metabolism , DNA Helicases , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Gene Expression Regulation, Viral , Vaccinia virus/genetics , Viral Proteins/genetics , Viral Proteins/metabolism , Animals , Protein Binding , Rabbits , Transcription Factors/genetics , Transcription Factors/metabolism , Transcription, Genetic , Vaccinia virus/metabolism
18.
Am Nat ; 152(4): 495-509, 1998 Oct.
Article in English | MEDLINE | ID: mdl-18811360

ABSTRACT

Forest ecologists often evaluate how well the species composition of saplings in the understory matches that of the canopy: absence of juveniles suggests that a tree species is suffering population decline. Here we offer a theoretical and empirical test of this assertion using data from a 50-ha census plot in Panama. Theory indicates that higher rates of population change, lambda, lead to more steeply declining size distributions (more juveniles relative to adults). But other parameters also affect the size distribution: lower growth rate of juveniles and lower survival at any size produce more steeply declining size distributions as well. Empirical evaluation of 216 tree populations showed that juvenile growth was the strongest predictor of size distribution, in the direction predicted by theory. Size distribution did correlate with population growth, but weakly and only in understory species, not canopy species. Size distribution did not correlate with the growth rate of larger individuals nor with survival. Results suggest that static in formation on the size distribution is not a good predictor of future population trends, while demographic information is. Fast-growing species will have fewer juveniles in the understory than slow growing species, even when population growth is equal.

19.
Virology ; 238(2): 391-409, 1997 Nov 24.
Article in English | MEDLINE | ID: mdl-9400612

ABSTRACT

We have previously reported the successful development of a targeted genetic method for the creation of temperature-sensitive vaccinia virus mutants [D. E. Hassett and R. C. Condit (1994) Proc. Natl. Acad. Sci. USA 91, 4554-4558]. This method has now been applied to the large subunit of the multifunctional vaccinia virus capping enzyme, encoded by gene D1R. Ten clustered charge-to-alanine mutations were created in a cloned copy of D1R. Four of these mutations were successfully transferred into the viral genome using transient dominant selection, and each of these four mutations yielded viruses with plaque phenotypes different from that of wild-type virus. Two of the mutant viruses, 516 and 793, were temperature sensitive in a plaque assay. Mutant 793 was also temperature sensitive in a one-step growth experiment. Phenotypic characterization of the 793 virus under both permissive and nonpermissive conditions revealed nearly normal patterns of viral protein and mRNA synthesis. Under nonpermissive conditions the 793 virus was defective in telomere resolution and blocked at an intermediate stage of viral morphogenesis. In vitro assays of various capping enzyme activities revealed that in permeabilized virions, enzyme guanylylate intermediate formation was reduced and methyltransferase activity was thermolabile, while in solubilized virion extracts enzyme guanylylate activity was reduced and both guanylyltransferase and methyltransferase activities were absent. Thus, the 793 mutation affects at least two separate enzymatic activities of the capping enzyme, guanylyltransferase and methyltransferase, and when incorporated into the virus genome, the mutation yields a virus that is temperature sensitive for growth, telomere resolution, and virion morphogenesis.


Subject(s)
Alanine , Methyltransferases/genetics , Multienzyme Complexes/genetics , Mutagenesis, Site-Directed , Nucleotidyltransferases/genetics , Phosphoric Monoester Hydrolases/genetics , Vaccinia virus/enzymology , Amino Acid Sequence , Cell Line , Methyltransferases/metabolism , Molecular Sequence Data , Morphogenesis , Multienzyme Complexes/metabolism , Nucleotidyltransferases/metabolism , Phenotype , Phosphoric Monoester Hydrolases/metabolism , Protein Processing, Post-Translational , RNA, Messenger/biosynthesis , RNA, Viral/biosynthesis , Telomere , Temperature , Vaccinia virus/genetics , Vaccinia virus/ultrastructure , Viral Plaque Assay , Viral Proteins/biosynthesis , Viral Proteins/metabolism , Virion
20.
Proc Natl Acad Sci U S A ; 94(4): 1252-7, 1997 Feb 18.
Article in English | MEDLINE | ID: mdl-11038601

ABSTRACT

Intraspecific density-dependent effects in the Barro Colorado Island (Panama) study area are far stronger, and involve far more species, than previously had been suspected. Significant effects on recruitment, many extremely strong, are seen for 67 out of the 84 most common species in the plot, including the 10 most common. Significant effects on the intrinsic rate of increase are seen in 54 of the 84 species. These effects are far more common than interspecific effects, and are predominantly of the type that should maintain tree diversity. As a result, the more diverse an area in the forest is, the higher is the overall rate of increase of the trees in that area, although sheer crowding has by itself a negative effect. These findings are consistent with, but do not prove, an important role for host-pathogen interactions (defined broadly) in the maintenance of diversity. Ways are suggested by which to test host-pathogen models and competing models.

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