Plant nuclear RNA polymerase IV mediates siRNA and DNA methylation-dependent heterochromatin formation

All eukaryotes have three nuclear DNA-dependent RNA polymerases, namely, Pol I, II, and III. Interestingly, plants have catalytic subunits for a fourth nuclear polymerase, Pol IV. Genetic and biochemical evidence indicates that Pol IV does not functionally overlap with Pol I, II, or III and is nonessential for viability. However, disruption of the Pol IV catalytic subunit genes NRPD1 or NRPD2 inhibits heterochromatin association into chromocenters, coincident with losses in cytosine methylation at pericentromeric 5S gene clusters and AtSN1 retroelements. Loss of CG, CNG, and CNN methylation in Pol IV mutants implicates a partnership between Pol IV and the methyltransferase responsible for RNA-directed de novo methylation. Consistent with this hypothesis, 5S gene and AtSN1 siRNAs are essentially eliminated in Pol IV mutants. The data suggest that Pol IV helps produce siRNAs that target de novo cytosine methylation events required for facultative heterochromatin formation and higher-order heterochromatin associations.     

Compensatory evolution of interacting gene products through multifunctional intermediates

When two mutations are singly deleterious but neutral or beneficial together, compensatory evolution can occur. The accumulation of derived, compensated genotypes contributes to the evolution of genetic incompatibilities between diverging populations or species. Previous two locus/two allele models have shown that compensatory evolution is appreciable only with tight linkage, the possibility of nearly simultaneous mutations, and/or a way to overcome negative selection against the singly mutated genotype. These conditions are often not met. Even when they are met, compensatory evolution is still predicted to be extremely slow, and in many scenarios selective advantage of the compensated genotype does little to accelerate it. Despite these obstacles, empirical studies suggest that it occurs readily. We describe here a set of related two locus/three allele models that invoke plausible neutral intermediates capable of productive interaction with both ancestral and compensated products of the interacting locus. These models are explored with analytical and computer simulation methods. The effect of these stepping-stone alleles on the evolution of ancestor-descendant incompatibilities is often profound, making the difference between evolution and stasis in several situations, including in small populations, when codominance or haploidy prevents shielding of mismatched genotypes, and in the absence of positive selection on the derived genotype. However, in large populations these intermediates can either speed or slow the evolution of incompatible genotypes relative to the two-allele case, depending on the specific fitness model. These results suggest that population size, the source of adaptive benefit, and the structural details of heteromeric gene product complexes interact to influence the path by which intergenic incompatibility evolves.     

Spatio-temporal patterns of the decline of freshwater mussels in the Little South Fork Cumberland River,USA

The Little South Fork Cumberland River, Kentucky and Tennessee, USA, was a globally important conservation refugium for freshwater mussels (Mollusca: Unionidae) because it supported an intact example (26 species) of the unique Cumberland River mussel fauna including imperiled species. We used previous surveys and our 1997–1998 survey to reconstruct the historical fauna, to describe spatio-temporal patterns of density and number of species, and to evaluate the probable sequence and cause of observed mussel declines. We were specifically interested in better understanding how mussel assemblages respond to chronic disturbances, and how these changes manifest in persistence patterns. Density and numbers of species declined steadily from 1981 to 1998, but declines occurred first in the lower river (early 1980s), followed by declines in the upper river (late 1980s to early 1990s). Of the total species recorded from the Little South Fork, 17 (65%) are seemingly extirpated and five others appear near extirpation. Declines are associated with at least two, temporally distinct major insults. Lower river declines are associated with surface mining, whereas, oil extraction activities are implicated in upper river declines. Regardless of causal factors, species persistence was primarily a function of predecline population size with only the most numerous and widespread species surviving. At this time, the river appears lost as a conservation refugium for mussels despite its remoteness, predominantly forested watershed, and several layers of existing statutory and regulatory environmental safeguards. We suggest that the river could be restored and mussels reintroduced if an interagency task force is formed to identify and mitigate specific stressors now affecting most mussel species in the river.Nomenclature: Turgeon et al. (1998).     

A candidate locus for variation in dispersal rate in a butterfly metapopulation

Frequent extinctions of local populations in metapopulations create opportunities for migrant females to establish new populations. In a metapopulation of the Glanville fritillary butterfly (Melitaea cinxia), more mobile individuals are more likely to establish new populations, especially in habitat patches that are poorly connected to existing populations. Here we show that flight metabolic rate and the frequency of a specific allele of the metabolic enzyme phosphoglucose isomerase (pgi) were both highest in newly established, isolated populations. Furthermore, genotypes with this pgi allele had elevated flight metabolic rates. These results suggest that genetic variation in pgi or a closely linked locus has a direct effect on flight metabolism, dispersal rate, and thereby on metapopulation dynamics in this species. These results also contribute to an emerging understanding of the mechanisms by which population turnover in heterogeneous landscapes may maintain genetic and phenotypic variation across populations.     

CD38 controls ADP-ribosyltransferase-2-catalyzed ADP-ribosylation of T cell surface proteins

ADP-ribosyltransferase-2 (ART2), a GPI-anchored, toxin-related ADP-ribosylating ectoenzyme, is prominently expressed by murine T cells but not by B cells. Upon exposure of T cells to NAD, the substrate for ADP-ribosylation, ART2 catalyzes ADP-ribosylation of the P2X7 purinoceptor and other functionally important cell surface proteins. This in turn activates P2X7 and induces exposure of phosphatidylserine and shedding of CD62L. CD38, a potent ecto-NAD-glycohydrolase, is strongly expressed by most B cells but only weakly by T cells. Following incubation with NAD, CD38-deficient splenocytes exhibited lower NAD-glycohydrolase activity and stronger ADP-ribosylation of cell surface proteins than their wild-type counterparts. Depletion of CD38(high) cells from wild-type splenocytes resulted in stronger ADP-ribosylation on the remaining cells. Similarly, treatment of total splenocytes with the CD38 inhibitor nicotinamide 2'-deoxy-2'-fluoroarabinoside adenine dinucleotide increased the level of cell surface ADP-ribosylation. Furthermore, the majority of T cells isolated from CD38-deficient mice "spontaneously" exposed phosphatidylserine and lacked CD62L, most likely reflecting previous encounter with ecto-NAD. Our findings support the notion that ecto-NAD functions as a signaling molecule following its release from cells by lytic or nonlytic mechanisms. ART2 can sense and translate the local concentration of ecto-NAD into corresponding levels of ADP-ribosylated cell surface proteins, whereas CD38 controls the level of cell surface protein ADP-ribosylation by limiting the substrate availability for ART2.     

Simultaneous differential staining of nucleic acids and proteins in histological tissues by means of J-Band effect

Summary A cationic carbocyanine dye, originally used in gel-electrophoresis, stains simultaneously DNA, RNA and proteins also in histological slides or smears. The resulting different colors are demonstrated by various spectral absorbance curves in cell nuclei and cytoplasm. The simple technique is suited also for quantitative cytophotometric determinations.The work was supported by the Deutsche Forschungsgemeinschaft.     

Single‐nucleotide polymorphisms of two closely related microsporidian parasites suggest a clonal population expansion after the last glaciation

The mode of reproduction of microsporidian parasites has remained puzzling since many decades. It is generally accepted that microsporidia are capable of sexual reproduction, and that some species have switched to obligate asexuality, but such process had never been supported with population genetic evidence. We examine the mode of reproduction of Hamiltosporidium tvaerminnensis and Hamiltosporidium magnivora, two closely related microsporidian parasites of the widespread freshwater crustacean Daphnia magna, based on a set of 129 single‐nucleotide polymorphisms distributed across 16 genes. We analyse 20 H. tvaerminnensis isolates from localities representative of the entire species' geographic distribution along the Skerry Island belt of the Baltic Sea. Five isolates of the sister species H. magnivora were used for comparison. We estimate the recombination rates in H. tvaerminnensis to be at least eight orders of magnitude lower than in H. magnivora and not significantly different from zero. This is corroborated by the higher divergence between H. tvaerminnensis alleles (including fixed heterozygosity), as compared to H. magnivora. Our study confirms that sexual recombination is present in microsporidia, that it can be lost, and that asexuals may become epidemic.     

Mating Damages the Cuticle of C. elegans Hermaphrodites

Lifespan costs to reproduction are common across multiple species, and such costs could potentially arise through a number of mechanisms. In the nematode Caenorhabditis elegans, it has been suggested that part of the lifespan cost to hermaphrodites from mating results from physical damage owing to the act of copulation itself. Here, we examine whether mating damages the surface of the hermaphrodite cuticle via scanning electron microscopy. It is found that mated hermaphrodites suffered delamination of cuticle layers surrounding the vulva, and that the incidence of such damage depends on genetic background. Unmated hermaphrodites demonstrated almost no such damage, even when cultured in soil with potentially abrasive particles. Thus, a consequence of mating for C. elegans hermaphrodites is physical cuticle damage. These experiments did not assess the consequences of cuticle damage for lifespan, and the biological significance of this damage remains unclear. We further discuss our results within the context of recent studies linking the lifespan cost to mating in C. elegans hermaphrodites to male secretions.