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1.
Mol Phylogenet Evol ; 55(2): 611-20, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20064622

RESUMO

The recognition of cryptic parasite species has implications for evolutionary and population-based studies of wildlife and human disease. Echinostome trematodes are a widely distributed, species-rich group of internal parasites that infect a wide array of hosts and are agents of disease in amphibians, mammals, and birds. We utilize genetic markers to understand patterns of morphology, host use, and geographic distribution among several species groups. Parasites from >150 infected host snails (Lymnaea elodes, Helisoma trivolvis and Biomphalaria glabrata) were sequenced at two mitochondrial genes (ND1 and CO1) and one nuclear gene (ITS) to determine whether cryptic species were present at five sites in North and South America. Phylogenetic and network analysis demonstrated the presence of five cryptic Echinostoma lineages, one Hypoderaeum lineage, and three Echinoparyphium lineages. Cryptic life history patterns were observed in two species groups, Echinostoma revolutum and Echinostoma robustum, which utilized both lymnaied and planorbid snail species as first intermediate hosts. Molecular evidence confirms that two species, E. revolutum and E. robustum, have cosmopolitan distributions while other species, E. trivolvis and Echinoparyphium spp., may be more geographically limited. The intra and interspecific variation detected in our study provides a genetic basis for seven species groups of echinostomes which will help accurately identify agents of disease as well as reveal cryptic aspects of trematode biology.


Assuntos
Echinostoma/genética , Filogenia , Caramujos/parasitologia , Animais , Núcleo Celular/genética , DNA de Helmintos/genética , DNA Mitocondrial/genética , Echinostoma/anatomia & histologia , Echinostoma/classificação , Geografia , América do Norte , Análise de Sequência de DNA , América do Sul
2.
BMC Genomics ; 10: 488, 2009 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-19845954

RESUMO

BACKGROUND: New chemotherapeutic agents against Schistosoma mansoni, an etiological agent of human schistosomiasis, are a priority due to the emerging drug resistance and the inability of current drug treatments to prevent reinfection. Proteases have been under scrutiny as targets of immunological or chemotherapeutic anti-Schistosoma agents because of their vital role in many stages of the parasitic life cycle. Function has been established for only a handful of identified S. mansoni proteases, and the vast majority of these are the digestive proteases; very few of the conserved classes of regulatory proteases have been identified from Schistosoma species, despite their vital role in numerous cellular processes. To that end, we identified protease protein coding genes from the S. mansoni genome project and EST library. RESULTS: We identified 255 protease sequences from five catalytic classes using predicted proteins of the S. mansoni genome. The vast majority of these show significant similarity to proteins in KEGG and the Conserved Domain Database. Proteases include calpains, caspases, cytosolic and mitochondrial signal peptidases, proteases that interact with ubiquitin and ubiquitin-like molecules, and proteases that perform regulated intramembrane proteolysis. Comparative analysis of classes of important regulatory proteases find conserved active site domains, and where appropriate, signal peptides and transmembrane helices. Phylogenetic analysis provides support for inferring functional divergence among regulatory aspartic, cysteine, and serine proteases. CONCLUSION: Numerous proteases are identified for the first time in S. mansoni. We characterized important regulatory proteases and focus analysis on these proteases to complement the growing knowledge base of digestive proteases. This work provides a foundation for expanding knowledge of proteases in Schistosoma species and examining their diverse function and potential as targets for new chemotherapies.


Assuntos
Biologia Computacional/instrumentação , Mineração de Dados , Genoma Helmíntico , Peptídeo Hidrolases/genética , Esquistossomose mansoni/genética , Animais , Etiquetas de Sequências Expressas , Genes de Helmintos , Filogenia , Esquistossomose mansoni/enzimologia
3.
Mol Ecol ; 18(15): 3307-15, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19508451

RESUMO

Major histocompatibility complex (MHC) alleles likely have adaptive value because of overdominance, in which case MHC heterozygous individuals have increased fitness relative to homozygotes. Because of this potential benefit, the evolution of sexual reproduction between MHC-divergent individuals (i.e. negative assortative mating, NAM) may be favoured. However, the strongest evidence for MHC-based NAM comes from inbred animals, and context-dependent mating preferences have rarely been evaluated although they often occur in nature. We assessed the extent MHC-based mating preferences among wild tiger salamanders (Ambystoma tigrinum) using multiple molecular approaches. We genotyped 102 adults and 864 larvae from 36 breeding trials at both microsatellite and MHC loci. Parentage analysis revealed that reproductive success among males was positively associated with increased tail length and that with respect to the focal female, MHC-similar males sired a significantly higher number of offspring than more dissimilar males. This trend was consistent, even under context-dependent scenarios that favour traditional MHC-based NAM. These results suggest that the most MHC-divergent males may be at a reproductive disadvantage in pairwise breeding trials. Our data add to a growing body of evidence that suggests where it exists, MHC-based choice is probably dynamic and mediated by many factors that vary in the wild, notably signals from other indicator traits and by the quality and quantity of potential mates.


Assuntos
Ambystoma/genética , Complexo Principal de Histocompatibilidade/genética , Preferência de Acasalamento Animal , Reprodução/genética , Ambystoma/anatomia & histologia , Animais , Feminino , Variação Genética , Genótipo , Masculino , Repetições de Microssatélites , Análise de Sequência de DNA
4.
Genetica ; 136(3): 501-4, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19107324

RESUMO

Microsatellites are commonly used for mapping and population genetics because of their high heterozygosities and allelic variability (i.e., polymorphism). Microsatellite markers are generally more polymorphic than other types of molecular markers such as allozymes or SNPs because the insertions/deletions that give rise to microsatellite variability are relatively common compared to nucleotide substitutions. Nevertheless, direct evidence of microsatellite mutation rates (MMRs) is lacking in most vertebrate groups despite the importance of such estimates to key population parameters (e.g., genetic differentiation or theta = 4N (e)micro). Herein, we present empirical data on MMRs in eastern tiger salamanders (Ambystoma tigrinum tigrinum). We conducted captive breeding trials and genotyped over 1,000 offspring at a suite of microsatellite loci. These data on 7,906 allele transfers provide the first direct estimates of MMRs in amphibians, and they illustrate that MMRs can vary by more than an order of magnitude across loci within a given species (one locus had ten mutations whereas the others had none).


Assuntos
Ambystoma/genética , Repetições de Microssatélites/genética , Mutagênese , Mutação/genética , Animais , Cruzamento , Genótipo , Modelos Genéticos , Polimorfismo Genético
5.
Biol Lett ; 4(5): 549-52, 2008 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-18593670

RESUMO

Inbreeding may lead to morphological malformations in a wide variety of taxa. We used genetic markers to evaluate whether malformed urodeles were more inbred and/or had less genetic diversity than normal salamanders. We captured 687 adult and 1,259 larval tiger salamanders (Ambystoma tigrinum tigrinum), assessed each individual for gross malformations, and surveyed genetic variation among malformed and normal individuals using both cytoplasmic and nuclear markers. The most common malformations in both adults and larvae were brachydactyly, ectrodactyly and polyphalangy. The overall frequency of adults with malformations was 0.078 compared to 0.081 in larval samples. Genetic diversity was high in both normal and malformed salamanders, and there were no significant difference in measures of inbreeding (f and F), allele frequencies, mean individual heterozygosity or mean internal relatedness. Environmental contaminants or other extrinsic factors may lead to genome alternations that ultimately cause malformations, but our data indicate that inbreeding is not a causal mechanism.


Assuntos
Ambystoma/anormalidades , Variação Genética , Endogamia , Deformidades Congênitas dos Membros/etiologia , Ambystoma/genética , Animais
6.
J Mol Evol ; 67(1): 68-75, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18543016

RESUMO

Alternative splicing (AS) of mRNA transcripts is increasingly recognized as a source of transcriptome diversity. To date, most AS studies have focused either on comparisons across taxa or on intragenomic comparisons across gene families. We generated a novel data set that represents one of the first population genetic comparisons of AS across individuals. In ambystomatid salamanders, AS of the major histocompatibility complex (MHC) class IIbeta gene (Amti-DAB) produces two transcripts, one full-length and one truncated. The full-length transcript is functional, but the truncated transcript is missing the critical beta1 domain that forms half of the peptide binding region in the intact MHC class II molecule. We captured wild salamander larvae (Ambystoma tigrinum tigrinum) and genotyped them at Amti-DAB via DNA sequencing. From these same larvae, we extracted RNA from gill and spleen and evaluated the relative expression level of Amti-DAB in each tissue. Across individuals, 21% of the transcripts were truncated (alternatively spliced), and the absolute level of alternative transcript expression was higher in gill. The high level of nucleotide variation among seven Amti-DAB alleles provides the ability to detect substitutions (or linked DNA polymorphisms) that might have influenced AS. The data reveal no correlation between AS and haplotype, allele, or zygosity. However, indirect evidence (comparative expression patterns across 3 million years of evolution) suggests that the truncated Amti-DAB transcript may be functional and maintained by natural selection.


Assuntos
Processamento Alternativo , Ambystoma/genética , Genes MHC da Classe II , Polimorfismo Genético , Alelos , Ambystoma/imunologia , Animais , Evolução Molecular , Genótipo , Antígenos de Histocompatibilidade Classe II/biossíntese , Antígenos de Histocompatibilidade Classe II/química , Modelos Moleculares , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , RNA Mensageiro/análise
7.
Evolution ; 62(6): 1458-68, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18331461

RESUMO

Microsatellites and mitochondrial DNA (mtDNA) have traditionally been used in population genetics because of their variability and presumed neutrality, whereas genes of the major histocompatibility complex (MHC) are increasingly of interest because strong selective pressures shape their standing variation. Despite the potential for MHC genes, microsatellites, and mtDNA sequences to complement one another in deciphering population history and demography, the three are rarely used in tandem. Here we report on MHC, microsatellite, and mtDNA variability in a single large population of the eastern tiger salamander (Ambystoma tigrinum tigrinum). We use the mtDNA mismatch distribution and, on microsatellite data, the imbalance index and bottleneck tests to infer aspects of population history and demography. Haplotype and allelic variation was high at all loci surveyed, and heterozygosity was high at the nuclear loci. We find concordance among neutral molecular markers that suggests our study population originated from post-Pleistocene expansions of multiple, fragmented sources that shared few migrants. Differences in N(e) estimates derived from haploid and diploid genetic markers are potentially attributable to secondary contact among source populations that experienced rapid mtDNA divergence and comparatively low levels of nuclear DNA divergence. We find strong evidence of natural selection acting on MHC genes and estimate long-term effective population sizes (N(e)) that are very large, making small selection intensities significant evolutionary forces in this population.


Assuntos
Ambystoma/genética , Variação Genética , Genética Populacional , Seleção Genética , Animais , Sequência de Bases , Primers do DNA/genética , DNA Mitocondrial/genética , Efeito Fundador , Haplótipos/genética , Complexo Principal de Histocompatibilidade/genética , Repetições de Microssatélites/genética , Dados de Sequência Molecular , Análise de Sequência de DNA
8.
Hereditas ; 144(6): 228-34, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18215245

RESUMO

The direct sequencing of PCR products from diploid organisms is problematic because of ambiguities associated with phase inference in multi-site heterozygotes. Several molecular methods such as cloning, SSCP, and DGGE have been developed to empirically reduce diploid sequences to their constitutive haploid components, but in theory these empirical approaches can be supplanted by analytical treatment of diploid sequences. Analytical approaches are more desirable than molecular methods because of the added time and expense required to generate molecular data. A variety of analytical methods have been developed to address this issue, but few have been rigorously evaluated with empirical data. Furthermore, they all assume that the sequences under consideration are evolving in a neutral fashion and assume a moderate number of heterozygous sites. Here, we use non-neutral major histocompatibility complex (MHC) sequences comprised of large numbers of heterozygous sites that are under strong balancing selection to evaluate the performance of the popular Bayesian algorithm implemented by the program PHASE. Our results suggest that PHASE performs admirably with non-neutral sequences of moderate length with numerous heterozygous sites typical of MHC class II sequences. We conclude that analytical approaches to haplotype inference have great potential in large-scale population genetic assays, but recommend groundtruthing analytical results using empirical (molecular) approaches at the outset of population-level analyses.


Assuntos
Genes MHC da Classe II , Genes MHC Classe I , Haplótipos , Complexo Principal de Histocompatibilidade/genética , Modelos Genéticos , Algoritmos , Animais , Teorema de Bayes , Mapeamento Cromossômico , Biologia Computacional/métodos , Método de Monte Carlo , Ploidias , Reprodutibilidade dos Testes , Salmão/genética , Urodelos/genética
9.
Mol Biol Evol ; 23(1): 137-43, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16162865

RESUMO

The patterns of major histocompatibility complex (MHC) evolution involve duplications, deletions, and independent divergence of loci during episodes punctuated by natural selection. Major differences in MHC evolution among taxa have previously been attributed to variation in linkage patterns of class I and class II MHC genes. Here we characterize patterns of evolution in the MHC class Ia gene of Xenopus laevis in terms of polymorphism, recombination, and extent of transspecies polymorphism. We also compare these patterns to see if a correlation exists with linkage or separation of the MHC class I and class II regions as seen in amphibians and teleost fishes. In X. laevis, we find high levels of polymorphism. Also, genetic exchange is relatively frequent and occurs in intron II, reshuffling allelic forms of exons 2 and 3. Evolutionary relationships among class I alleles show an intermingling of alleles from divergent Xenopus species rather than a species-specific clustering. Results indicate that the patterns of evolution are similar to those found in salmonid fishes and are different from the mode of evolution seen in primates. Similar patterns of class Ia evolution in salmonid fishes and X. laevis suggest that nonlinkage of class I and class II regions alone is insufficient to explain some patterns of MHC evolution in salmonids.


Assuntos
Evolução Molecular , Genes MHC Classe I/genética , Filogenia , Polimorfismo Genético , Recombinação Genética , Xenopus laevis/genética , Animais , Sequência de Bases , Primers do DNA , Funções Verossimilhança , Modelos Genéticos , Dados de Sequência Molecular , Análise de Sequência de DNA
10.
Immunogenetics ; 57(10): 775-81, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16195876

RESUMO

Major histocompatibility complex (MHC) class II genes are usually among the most polymorphic in vertebrate genomes because of their critical role (antigen presentation) in immune response. Prior to this study, the MHC was poorly characterized in tiger salamanders (Ambystoma tigrinum), but the congeneric axolotl (Ambystoma mexicanum) is thought to have an unusual MHC. Most notably, axolotl class II genes lack allelic variation and possess a splice variant without a full peptide binding region (PBR). The axolotl is considered immunodeficient, but it is unclear how or to what extent MHC genetics and immunodeficiency are interrelated. To study the evolution of MHC genes in urodele amphibians, we describe for the first time an expressed polymorphic class II gene in wild tiger salamanders. We sequenced the PBR of a class II gene from wild A. tigrinum (n=33) and identified nine distinct alleles. Observed heterozygosity was 73%, and there were a total of 46 polymorphic sites, most of which correspond to amino acid positions that bind peptides. Patterns of nucleotide substitutions exhibit the signature of diversifying selection, but no recombination was detected. Not surprisingly, trans-species evolution of tiger salamander and axolotl class II alleles was apparent. We have no direct data on the immunodeficiency of tiger salamanders, but the levels of polymorphism in our study population should suffice to bind a variety of foreign peptides (unlike axolotls). Our tiger salamander data suggest that the monomorphism and immunodeficiencies associated with axolotl class II genes is a relic of their unique historical demography, not their phylogenetic legacy.


Assuntos
Ambystoma/imunologia , Antígenos de Histocompatibilidade Classe II/genética , Polimorfismo Genético , Alelos , Ambystoma/genética , Sequência de Aminoácidos , Animais , Dados de Sequência Molecular , Filogenia , Homologia de Sequência de Aminoácidos
11.
J Mol Evol ; 60(2): 221-8, 2005 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15785850

RESUMO

The LMP7 and PSMB5 genes were created through an ancient gene duplication event of their ancestral locus. These proteins contain an active site of proteolysis, and LMP7 replaces PSMB5 as a component of the 20S proteasome after stimulation of cells by interferon-gamma. Replacement of PSMB5 by LMP7 changes the profile of the products of 20S proteasome processing, predisposing digested peptides for transport to and display by the immune system. The purpose of this study is to investigate evolutionary forces influencing functional divergence between LMP7 and PSMB5 following duplication. Levels of synonymous and nonsynonymous substitution rates are estimated to infer differences in levels of natural selection. Estimates of substitution rates indicate that natural selection elevated rates of nonsynonymous substitution in LMP7 following gene duplication, whereas PSMB5 experienced an increase in substitution rate that was not likely due to diversifying natural selection following duplication. Following initial divergence, nearly neutral mutations have dominated gene evolution in both lineages. The LMP7 gene locus provides a rare example of a protein with specialized function arising from duplication and divergence of a housekeeping protein by way of natural selection.


Assuntos
Duplicação Gênica , Complexos Multienzimáticos/genética , Complexo de Endopeptidases do Proteassoma/genética , Seleção Genética , Animais , DNA/genética , Evolução Molecular , Humanos , Modelos Genéticos , Complexos Multienzimáticos/química , Filogenia , Complexo de Endopeptidases do Proteassoma/química , Subunidades Proteicas
12.
Dev Comp Immunol ; 29(3): 211-27, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15572070

RESUMO

Molecular phylogenetics and its applications are popular and useful tools for making comparative investigations in genetics; however, estimating phylogenetic trees is not always straightforward. Some phylogenetic estimators use an explicit model of nucleotide evolution to estimate evolutionary parameters such as branch lengths and tree topology. There are many models to choose from, and use of the optimal model for a particular data set is important to avoid a loss of power and accuracy in phylogenetic estimations. Here, we review some molecular evolutionary forces and the parameters included in some common models of evolution used to interpret resulting patterns of molecular variation. We present some statistical methods of selecting a particular model of nucleotide evolution, and provide an empirical example of model selection. Statistical model selection strikes a balance between the bias introduced by some models and the increased variance of parameter estimates that results from using other models.


Assuntos
Evolução Molecular , Modelos Genéticos , Filogenia , Animais , Teorema de Bayes , Humanos , Funções Verossimilhança
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