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The organization of the evolutionarily conserved GATA/GACA repeats in the mouse genome.

Simple repeated GATA and GACA sequences which were originally isolated from sex-specific snake satellite DNA have been found subsequently in all eukaryotes studied. The organization of these sequences within the mouse genome was investigated here by using synthetic oligonucleotide probes as a novel tool in comparison with conventional hybridization probes. Southern blot hybridization showed sex-specific patterns with both the (GATA)4 and (GACA)4 oligonucleotide probes, as previously described with conventional probes. The quantitative analysis of two mouse DNA phage libraries and of 25 isolated GATA-positive phage clones revealed intensive interspersion of GATA sequences with GACA, and with other repetitive and single-copy sequences. Ubiquitous interspersion and homogeneous genomic distribution of GATA and GACA sequences were confirmed by hybridization in situ of the oligonucleotide probes to metaphase chromosomes. The lengths of the GATA and GACA stretches were found to vary considerably in the individual phage clones. DNA inserts from 20 phages were assigned to autosomes and sex chromosomes and three genomic fragments were found to be confined to the Y chromosome. The organization of GATA and GACA sequences is discussed in the context of their evolutionary potential and possible conservation mechanisms.

Animals↗

Quantitative-genetic analysis of wing form and bilateral asymmetry in isochromosomal lines of Drosophila subobscura using Procrustes methods.

Fluctuating asymmetry (FA) is often used as a measure of underlying developmental instability (DI), motivated by the idea that morphological variance is maladaptive. Whether or not DI has evolutionary potential is a highly disputed topic, marred by methodological problems and fuzzy prejudices. We report here some results from an ongoing study of the effects of karyotype, homozygosity and temperature on wing form and bilateral asymmetry using isochromosomal lines of Drosophila subobscura. Our approach uses the recently developed methodologies in geometric morphometrics to analyse shape configurations of landmarks within the standard statistical framework employed in studies of bilateral asymmetries, and we have extended these methods to partition the individual variation and the variation in asymmetries into genetic and environmental causal components. The analyses revealed temperature-dependent expression of genetic variation for wing size and wing shape, directional asymmetry (DA) of wing size, increased asymmetries at suboptimal temperature, and a transition from FA to DA in males as a result of increase in the rearing temperature. No genetic variation was generally detected for FA in our samples, but these are preliminary results because no crosses between lines were carried out and, therefore, the contribution of dominance was not taken into account. In addition, only a subset of the standing genetic variation was represented in the experiments.

Animals↗

The behavior of a Daphnia pulex transposable element in cyclically and obligately parthenogenetic populations.

Using Southern blot analysis, we have characterized restriction fragment patterns of a transposable element. Pokey, in obligately and cyclically parthenogenetic populations of the cladoceran crustacean Daphnia pulex. We show that the element is most likely active in cyclically parthenogenetic populations but is, for the most part, inactive in obligate parthenogens. This result is consistent with theory suggesting that transposable element dynamics are likely to change with a change in reproductive mode. Such changes could have important consequences for the long-term evolutionary potential of obligate parthenogens and may also be informative with regard to the underlying mechanisms that regulate transposable element frequencies in sexual organisms.

Animals↗

Polymorphism of major ribosomal gene chromosomal sites (NOR-phenotypes) in the hybridogenetic fish Squalius alburnoides complex (Cyprinidae) assessed through crossing experiments.

Chromosomal locations of major ribosomal sites, i.e. NOR-phenotypes, were assigned in Squalius alburnoides complex using sequential chromomycin A3 (CMA3)- and silver (Ag)-staining. This hybridogenetic Iberian minnow comprises diploid, triploid and tetraploid forms that arose by interspecific hybridisation between S. pyrenaicus and an unknown species. Inheritance of NOR patterns was studied by means of crossing experiments involving most diploid-polyploid forms of the S. alburnoides complex with identified specific genotype constitution. In all the specimens studied, the NORs were localised in the short arms of submetacentric chromosomes. Although S. pyrenaicus presented only one pair of NOR-bearing chromosomes, the data from experimental crosses evidenced that S. alburnoides complex was characterised by a multiple NOR phenotype composed of one chromosome pair with stable NORs and two chromosome pairs with NOR site polymorphism of presence/absence type. These data suggest that the karyotype of the unknown parental species of the S. alburnoides complex should have a multiple NOR pattern and emphasised the role of the all-male diploid linage in the dynamics and evolutionary potential of the S. alburnoides complex allowing the preservation of the missing ancestor genome. Cross-analyses evidenced that in spite of the high polymorphic nature of NORs in this fish complex, we have no reason to reject the hypothesis that their inheritance patterns were in accordance with Mendelian segregation.

Animals↗

Brave New World: the epistatic foundations of natives adapting to invaders.

Classical examples indicated rapid evolution to be both rare and largely anthropogenic. As the pace and scale of human disturbance increase, such evolution is becoming more the norm. Genetically based adaptation may underlie successful biological invasions, and may likewise characterize responses in natives to invasives. Recent published studies confirm that natives are adapting morphologically, behaviorally, physiologically and life historically to selection from invasive species. Some of the processes involved are evident in our studies of recent host shifts to invasive plants by native soapberry bugs in North America and Australia. On both continents populations have differentiated extensively in fitness traits. Genetic architecture of these adaptations involves a surprising degree of non-additive variation (epistasis, dominance), a result that in theory may reflect a history of colonization by a small number of individuals followed by population growth. Such "founder-flush" events may unleash extraordinary evolutionary potential, and their importance will be clarified as more studies take advantage of the accidental perturbation experiments that biotic invasions represent. From a conservation standpoint, rapid evolution in natives will present challenges for ecologically appropriate and sustainable management, but at the same time may enhance the capacity of the native community to act in the biological control of invasive species.

Adaptation, Physiological↗

Identification of previously unrecognized sequence motifs at the extreme carboxyterminus of the neurofilament subunit NF-M.

Two previously unrecognized features of neurofilament architecture are revealed by careful analysis of published neurofilament sequences. 1. The extreme C-terminus of the NF-M tail contains two highly conserved homologous sequences each of 15 amino acids, with the consensus EEK-V-TKKVEK-TS, plus another very closely related 7 amino acid sequence. 2. The C-terminus of NF-M contains sequences of consensus K-SP or K--SP which in some species are multiply repeated, are probably phosphorylated, but are distinct from the more obvious KSP repeated sequences. Sequences related to both the K-SP and K--SP sequences are found in NF-H, microtubule associated proteins tau and MAP2, suggesting a further level of immunological and potential evolutionary relationship between neurofilaments and these microtubule associated proteins. The possible significance of these findings is discussed.

Amino Acid Sequence↗

Structure of cryptic lambda prophages.

When Escherichia coli cells lysogenic for bacteriophage lambda are induced with ultraviolet light, cells carrying cryptic lambda prophages are occasionally found among the apparently cured survivors. The lambda variant crypticogen (lambda crg) carries an insertion of the transposable element IS2, which increases the frequency of cryptic lysogens to about 50% of cured cells: 43 of these cryptic prophages have been characterized. They all contain substitutions that replace the early segment of the prophage genome (from the IS2 to near the cos site) with a duplicate copy of a large segment of the host chromosome. The right end of the substitution always results from recombination between the nin-QSR-cos region of the prophage and the homologous incomplete lambdoid prophage Qsr' at 12.5 minutes in the E. coli chromosome. The left end of the substitution is usually a crossover that recombines the IS2 element in the prophage with an E. coli IS2 at 8.5 minutes, near the lac gene, or with a second IS2 located counterclockwise from leu at 2 minutes, generating duplications of at least 200,000 bases. Five cryptic lysogens derived from cells lysogenic for a reference strain of lambda (which lacks the IS2 present in lambda crg) have been characterized. They contain substitutions whose right termini are generated by a crossover with the Qsr' prophage. The left termini of these substitutions are formed either by a crossover between the lambda exo gene and a short exo-homologous segment of Qsr' (2/5), or by a crossover between sequences to the left of attL and an unmapped distant region of the host chromosome (3/5). The large duplications carried by these cryptic lysogens are stable, unlike tandem duplications, and so may significantly influence the cell's evolutionary potential.

Bacteriophage lambda↗

The catadromous European eel Anguilla anguilla (L.) as a model for freshwater evolutionary ecotoxicology: relationship between heavy metal bioaccumulation, condition and genetic variability.

Understanding the effects of pollutants on the genome is of crucial importance to preserve the evolutionary potential of endangered natural populations. The highly vagile European eel (Anguilla anguilla L.) has suffered a dramatic decline in recruitment since two decades, urging for a better understanding of the genetic impact of pollution. Its catadromous life history constitutes a model to assess local selection of pollutants on condition and genetic variability, as juveniles recruit in European rivers without appreciable pollution load or interfering genetic background. Because of its high fat content and local benthic feeding behaviour, the feeding stage is considered extremely prone to the bioaccumulation of pollutants. We studied the relationship between heavy metal bioaccumulation, fitness (condition) and genetic variability in the European eel. The muscle tissues of 78 sub-adult eels, originating from three Belgian river basins (Scheldt, Meuse and Yser), were examined for nine heavy metal pollutants (Hg, Cd, Pb, Cu, Zn, Ni, Cr, As and Se), while in total 123 individuals were genotyped at 12 allozyme and 8 microsatellite loci. A significant negative correlation between heavy metal pollution load and condition was observed, suggesting an impact of pollution on the health of sub-adult eels. In general, we observed a reduced genetic variability in strongly polluted eels, as well as a negative correlation between level of bioaccumulation and allozymatic multi-locus heterozygosity (MLH). Microsatellite genetic variability did not show any pollution related differences, suggesting a differential response at metabolic enzymes and possibly direct overdominance of heterozygous individuals.

Anguilla↗

Specific dynamic action: a century of investigation.

Specific dynamic action (SDA) is the term used to refer to the increased metabolic expenditure that occurs in postprandial animals. Postprandial increases in metabolism were first documented in animals over two hundred years ago, and have since been observed in every species thus far examined. Ironically, the ubiquity of this physiological response to feeding understates its complex nature. This review is designed to summarize both classical and modern hypotheses regarding the causality of SDA as well as to review important findings from the past century of scientific research into SDA. A secondary aim of this work is to emphasize the importance of carefully designed experiments and systematic hypothesis testing to make more rapid progress in understanding the physiological processes that contribute to SDA. I also identify three areas in SDA research that deserve more detailed investigation. The first area is identification of the causality of SDA in 'model' organisms. The second area is characterization of SDA responses in novel species. The third area is exploration of the ecological and potential evolutionary significance of SDA in energy budgets of animals.

Animals↗

Contrasting views over a hybrid complex: Between speciation and evolutionary "dead-end".

The Squalius alburnoides complex, of hybrid origin, comprises diploid, triploid and tetraploid forms and has a widespread distribution in the Iberian waters. The southern populations of this complex, sympatric with S. pyrenaicus, show high genetic variability, diversity of forms and reproductive modes which create pathways that may allow for the establishment of a new species in the future. Here we report a contrasting view over the S. alburnoides complex: in the Mondego River basin (northern Portugal), nuclear "non-hybrid" and tetraploid forms are absent and a clearly impoverished genetic diversity is observed, contributing to a general scarcity of possibilities of generating novel genetic material. Moreover, the bisexual species involved in the maintenance of the complex in this basin (S. carolitertii) exhibits a considerably lower genetic variability, when compared with S. pyrenaicus. The observed differences suggest that, despite being originated by similar hybridization events and maintained by analogous reproductive mechanisms, different populations of the complex were exposed to distinct evolutionary constrains, which in some cases resulted in diversification and speciation while in others led to a compromising situation in terms of evolutionary potential. Additionally, and for the first time all forms were used in the calculation of genetic distances and diversity indices, widening the possibilities of analysis of the complex by allowing the inclusion of a large part of the available data, irrespective of ploidy level.

Animals↗

Analysis of Sarcoptes scabiei finds no evidence of infection with Wolbachia.

The endosymbiont Wolbachia has been detected in a range of filarial nematodes and parasitic mites and is known to affect host reproductive compatibility and potentially evolutionary processes. PCR of Wolbachia surface protein (wsp), ftsZ and 16SrRNA genes from individual Sarcoptes scabiei mites obtained from a series of individual hosts, and database searches of an S. scabiei var. hominis EST library failed to detect Wolbachia genes. Therefore, Wolbachia appears not to be involved in the genetic subdivision observed between varieties of host-associated S. scabiei or, involved in the inflammatory disease pathogenesis of scabies unlike its activity in filarial infection.

Animals↗

Tracking the evolution of porphobilinogen synthase metal dependence in vitro.

Metal ions are indispensable cofactors for chemical catalysis by a plethora of enzymes. Porphobilinogen synthases (PBGSs), which catalyse the second step of tetrapyrrole biosynthesis, are grouped according to their dependence on Zn(2+). Using site-directed mutagenesis, we embarked on transforming Zn(2+)-independent Pseudomonas aeruginosa PBGS into a Zn(2+)-dependent enzyme. Nine PBGS variants were generated by permutationally introducing three cysteine residues and a further two residues into the active site of the enzyme to match the homologous Zn(2+)-containing PBGS from Escherichia coli. Crystal structures of seven enzyme variants were solved to elucidate the nature of Zn(2+) coordination at high resolution. The three single-cysteine variants were invariably found to be enzymatically inactive and only one (D139C) was found to bind detectable amounts of Zn(2+). The double mutant A129C/D139C is enzymatically active and binds Zn(2+) in a tetrahedral coordination. Structurally and functionally it mimics mycobacterial PBGS, which bears an equivalent Zn(2+)-coordination site. The remaining two double mutants, without known natural equivalents, reveal strongly distorted tetrahedral Zn(2+)-binding sites. Variant A129C/D131C possesses weak PBGS activity while D131C/D139C is inactive. The triple mutant A129C/D131C/D139C, finally, displays an almost ideal tetrahedral Zn(2+)-binding geometry and a significant Zn(2+)-dependent enzymatic activity. Two additional amino acid exchanges further optimize the active site architecture towards the E.coli enzyme with an additional increase in activity. Our study delineates the potential evolutionary path between Zn(2+)-free and Zn(2+)-dependent PBGS enyzmes showing that the rigid backbone of PBGS enzymes is an ideal framework to create or eliminate metal dependence through a limited number of amino acid exchanges.

Amino Acid Sequence↗

[Neuroendocrine tumours treated with mIBG and radiolabeled peptides].

Neuroendocrine tumours are a heterogeneous group, characterized by good prognosis, but important disparities of the evolutionary potential. In the aggressive forms, the therapeutic strategies are limited. The metabolic or systemic radiotherapy, using radiolabelled peptides, which can act at the same time on the primary tumour and its metastases, constitutes a tempting therapeutic alternative, currently in evolution. The preliminary results are encouraging; the prospects are related to the development of new radiopharmaceuticals, with the use of other peptide analogues whose applications will overflow the framework of the neuroendocrine tumours.

3-Iodobenzylguanidine↗

Waste not, want not--transcript excess in multicellular eukaryotes.

There is growing evidence that mammalian genomes produce thousands of transcripts that do not encode proteins, and this RNA class might even rival the complexity of mRNAs. There is no doubt that a number of these non-protein-coding RNAs have important regulatory functions in the cell. However, do all transcripts have a function or are many of them products of fortuitous transcription with no function? The second scenario is mirrored by numerous alternative-splicing events that lead to truncated proteins. Nevertheless, analogous to 'superfluous' genomic DNA, aberrant transcripts or processing products embody evolutionary potential and provide novel RNAs that natural selection can act on.

Alternative Splicing↗

Analysis of the functional relationship between V3 loop and gp120 context with regard to human immunodeficiency virus coreceptor usage using naturally selected sequences and different viral backbones.

The human immunodeficiency virus type 1 (HIV-1) gp120 V3 loop plays a predominant role in chemokine receptor usage; however, other linear and nonlinear gp120 domains are involved in this step of the HIV-1 replication cycle. At present, the functional relationship between V3 and these domains with regard to coreceptor usage is unclear. To gain insights into the nature of this relationship in naturally selected viral variants, we developed a recombinant strategy based on two different gp120 backbones derived from CXCR4 (X4)- and CCR5 (R5)-tropic viral strains, respectively. Using this recombinant model system, we evaluated the phenotype patterns conferred to chimeric viruses by exogenous V3 loops from reference molecular clones and samples from infected subjects. In 13 of 17 recombinants (76%), a comparable phenotype was observed independently of the gp120 backbone, whereas in a minority of the recombinant viruses (4/17, 24%) viral infectivity depended on the gp120 context. No case of differential tropism using identical V3 sequence in the two gp120 contexts was observed. Site-directed mutagenesis experiments were performed to evaluate the phenotypic impact of specific V3 motifs. The data indicate that while the interaction of HIV-1 with chemokine receptors is driven by V3 loop and influenced by its evolutionary potential, the gp120 context plays a role in influencing the replication competence of the variants, suggesting that compensatory mutations occurring at sites other than V3 are necessary in some cases.

Amino Acid Sequence↗

Fitness landscapes and the myeloid leukemias.

Fitness landscapes, which provide a unique perspective for viewing co-evolving cell populations, were used to study the evolution of CML and MDS. This led to several conclusions: (1) accelerated phase CML and RAEB/RAEBt are not specific disease entities. They represent the time when AML cells are replacing preleukemia cells; (2) monoclonal hemopoiesis and RA/RARS represent a variety of clinical syndromes with a common appearance but with different evolutionary potential; (3) malignant cells alter the fitness landscape enhancing their proliferative advantage. These studies provide the basis for new approaches to treatment.

Anemia, Refractory, with Excess of Blasts↗

Using HIV-1 sequence variability to explore virus biology.

Human immunodeficiency virus type 1 (HIV-1) only recently established an epidemic world-wide infection in the human population. The virus persists in the human host through active replication and is able to avoid clearance by the immune system. Active replication is an important component of the rapid evolutionary potential of HIV-1, a potential which manifests itself in the evolution of immune escape variants, drug resistant variants, and variants with the ability to use different cell surface coreceptors in conjunction with CD4. Multiple zoonotic introductions, compartmentalization of virus replication in the body, and genetic bottlenecks associated with sampling during transmission, antiretroviral therapy, and geographic and/or host population isolation further contribute to the range of sequences present in extant viruses. The sum of the history of all of these phenomena is reflected in HIV-1 sequence variability, and most of these phenomena are ongoing today. Here we review the use of HIV-1 sequence variability to explore its underlying biology.

Animals↗

What good is it to get old?

The physiologic basis for aging is still incompletely understood, and most investigators have looked at aging as a process of decay without any clear adaptive advantage to the species. It is proposed that aging has potential evolutionary advantages, and that the aging process may have been the subject of positive selection.

Aged↗