Chromatin and siRNA pathways cooperate to maintain DNA methylation of small transposable elements in Arabidopsis

BACKGROUND: DNA methylation occurs at preferred sites in eukaryotes. In Arabidopsis, DNA cytosine methylation is maintained by three subfamilies of methyltransferases with distinct substrate specificities and different modes of action. Targeting of cytosine methylation at selected loci has been found to sometimes involve histone H3 methylation and small interfering (si)RNAs. However, the relationship between different cytosine methylation pathways and their preferred targets is not known. RESULTS: We used a microarray-based profiling method to explore the involvement of Arabidopsis CMT3 and DRM DNA methyltransferases, a histone H3 lysine-9 methyltransferase (KYP) and an Argonaute-related siRNA silencing component (AGO4) in methylating target loci. We found that KYP targets are also CMT3 targets, suggesting that histone methylation maintains CNG methylation genome-wide. CMT3 and KYP targets show similar proximal distributions that correspond to the overall distribution of transposable elements of all types, whereas DRM targets are distributed more distally along the chromosome. We find an inverse relationship between element size and loss of methylation in ago4 and drm mutants. CONCLUSION: We conclude that the targets of both DNA methylation and histone H3K9 methylation pathways are transposable elements genome-wide, irrespective of element type and position. Our findings also suggest that RNA-directed DNA methylation is required to silence isolated elements that may be too small to be maintained in a silent state by a chromatin-based mechanism alone. Thus, parallel pathways would be needed to maintain silencing of transposable elements

Sequences Sufficient for Programming Imprinted Germline DNA Methylation Defined

Epigenetic marks are fundamental to normal development, but little is known about signals that dictate their placement. Insights have been provided by studies of imprinted loci in mammals, where monoallelic expression is epigenetically controlled. Imprinted expression is regulated by DNA methylation programmed during gametogenesis in a sex-specific manner and maintained after fertilization. At Rasgrf1 in mouse, paternal-specific DNA methylation on a differential methylation domain (DMD) requires downstream tandem repeats. The DMD and repeats constitute a binary switch regulating paternal-specific expression. Here, we define sequences sufficient for imprinted methylation using two transgenic mouse lines: One carries the entire Rasgrf1 cluster (RC); the second carries only the DMD and repeats (DR) from Rasgrf1. The RC transgene recapitulated all aspects of imprinting seen at the endogenous locus. DR underwent proper DNA methylation establishment in sperm and erasure in oocytes, indicating the DMD and repeats are sufficient to program imprinted DNA methylation in germlines. Both transgenes produce a DMD-spanning pit-RNA, previously shown to be necessary for imprinted DNA methylation at the endogenous locus. We show that when pit-RNA expression is controlled by the repeats, it regulates DNA methylation in cis only and not in trans. Interestingly, pedigree history dictated whether established DR methylation patterns were maintained after fertilization. When DR was paternally transmitted followed by maternal transmission, the unmethylated state that was properly established in the female germlines could not be maintained. This provides a model for transgenerational epigenetic inheritance in mice

Quantifying the effect of forest age in annual net forest carbon balance

Forests dominate carbon (C) exchanges between the terrestrial biosphere and the atmosphere on land. In the long term, the net carbon flux between forests and the atmosphere has been significantly impacted by changes in forest cover area and structure due to ecological disturbances and management activities. Current empirical approaches for estimating net ecosystem productivity (NEP) rarely consider forest age as a predictor, which represents variation in physiological processes that can respond differently to environmental drivers, and regrowth following disturbance. Here, we conduct an observational synthesis to empirically determine to what extent climate, soil properties, nitrogen deposition, forest age and management influence the spatial and interannual variability of forest NEP across 126 forest eddy-covariance flux sites worldwide. The empirical models explained up to 62% and 71% of spatio-temporal and across-site variability of annual NEP, respectively. An investigation of model structures revealed that forest age was a dominant factor of NEP spatio-temporal variability in both space and time at the global scale as compared to abiotic factors, such as nutrient availability, soil characteristics and climate. These findings emphasize the importance of forest age in quantifying spatio-temporal variation in NEP using empirical approaches

ZBED6, a Novel Transcription Factor Derived from a Domesticated DNA Transposon Regulates IGF2 Expression and Muscle Growth

This study identifies a previously uncharacterized protein, encoded by a domesticated DNA transposon, called ZBED6 that regulates the expression of the insulin-like growth factor 2 (IGF2) gene, and possibly numerous others, in all placental mammals including human

Methane exchange in a boreal forest estimated by gradient method

Forests are generally considered to be net sinks of atmospheric methane (CH4) because of oxidation by methanotrophic bacteria in well-aerated forests soils. However, emissions from wet forest soils, and sometimes canopy fluxes, are often neglected when quantifying the CH4 budget of a forest. We used a modified Bowen ratio method and combined eddy covariance and gradient methods to estimate net CH4 exchange at a boreal forest site in central Sweden. Results indicate that the site is a net source of CH4. This is in contrast to soil, branch and leaf chamber measurements of uptake of CH4. Wetter soils within the footprint of the canopy are thought to be responsible for the discrepancy. We found no evidence for canopy emissions per se. However, the diel pattern of the CH4 exchange with minimum emissions at daytime correlated well with gross primary production, which supports an uptake in the canopy. More distant source areas could also contribute to the diel pattern; their contribution might be greater at night during stable boundary layer conditions

Antagonism between DNA and H3K27 Methylation at the Imprinted Rasgrf1 Locus

At the imprinted Rasgrf1 locus in mouse, a cis-acting sequence controls DNA methylation at a differentially methylated domain (DMD). While characterizing epigenetic marks over the DMD, we observed that DNA and H3K27 trimethylation are mutually exclusive, with DNA and H3K27 methylation limited to the paternal and maternal sequences, respectively. The mutual exclusion arises because one mark prevents placement of the other. We demonstrated this in five ways: using 5-azacytidine treatments and mutations at the endogenous locus that disrupt DNA methylation; using a transgenic model in which the maternal DMD inappropriately acquired DNA methylation; and by analyzing materials from cells and embryos lacking SUZ12 and YY1. SUZ12 is part of the PRC2 complex, which is needed for placing H3K27me3, and YY1 recruits PRC2 to sites of action. Results from each experimental system consistently demonstrated antagonism between H3K27me3 and DNA methylation. When DNA methylation was lost, H3K27me3 encroached into sites where it had not been before; inappropriate acquisition of DNA methylation excluded normal placement of H3K27me3, and loss of factors needed for H3K27 methylation enabled DNA methylation to appear where it had been excluded. These data reveal the previously unknown antagonism between H3K27 and DNA methylation and identify a means by which epigenetic states may change during disease and development

The biophysical climate mitigation potential of boreal peatlands during the growing season

Peatlands and forests cover large areas of the boreal biome and are critical for global climate regulation. They also regulate regional climate through heat and water vapour exchange with the atmosphere. Understanding how land-atmosphere interactions in peatlands differ from forests may therefore be crucial for modelling boreal climate system dynamics and for assessing climate benefits of peatland conservation and restoration. To assess the biophysical impacts of peatlands and forests on peak growing season air temperature and humidity, we analysed surface energy fluxes and albedo from 35 peatlands and 37 evergreen needleleaf forests-the dominant boreal forest type-and simulated air temperature and vapour pressure deficit (VPD) over hypothetical homogeneous peatland and forest landscapes. We ran an evapotranspiration model using land surface parameters derived from energy flux observations and coupled an analytical solution for the surface energy balance to an atmospheric boundary layer (ABL) model. We found that peatlands, compared to forests, are characterized by higher growing season albedo, lower aerodynamic conductance, and higher surface conductance for an equivalent VPD. This combination of peatland surface properties results in a similar to 20% decrease in afternoon ABL height, a cooling (from 1.7 to 2.5 degrees C) in afternoon air temperatures, and a decrease in afternoon VPD (from 0.4 to 0.7 kPa) for peatland landscapes compared to forest landscapes. These biophysical climate impacts of peatlands are most pronounced at lower latitudes (similar to 45 degrees N) and decrease toward the northern limit of the boreal biome (similar to 70 degrees N). Thus, boreal peatlands have the potential to mitigate the effect of regional climate warming during the growing season. The biophysical climate mitigation potential of peatlands needs to be accounted for when projecting the future climate of the boreal biome, when assessing the climate benefits of conserving pristine boreal peatlands, and when restoring peatlands that have experienced peatland drainage and mining.Peer reviewe

Quantifying the effect of forest age in annual net forest carbon balance

Forests dominate carbon (C) exchanges between the terrestrial biosphere and the atmosphere on land. In the long term, the net carbon flux between forests and the atmosphere has been significantly impacted by changes in forest cover area and structure due to ecological disturbances and management activities. Current empirical approaches for estimating net ecosystem productivity (NEP) rarely consider forest age as a predictor, which represents variation in physiological processes that can respond differently to environmental drivers, and regrowth following disturbance. Here, we conduct an observational synthesis to empirically determine to what extent climate, soil properties, nitrogen deposition, forest age and management influence the spatial and interannual variability of forest NEP across 126 forest eddy-covariance flux sites worldwide. The empirical models explained up to 62% and 71% of spatio-temporal and across-site variability of annual NEP, respectively. An investigation of model structures revealed that forest age was adominant factor of NEP spatio-temporal variability in both space and time at the global scale as compared to abiotic factors, such as nutrient availability, soil characteristics and climate. These findings emphasize the importance of forest age in quantifying spatio-temporal variation in NEP using empirical approaches

Globally altered epigenetic landscape and delayed osteogenic differentiation in H3.3-G34W-mutant giant cell tumor of bone

Abstract The neoplastic stromal cells of giant cell tumor of bone (GCTB) carry a mutation in H3F3A, leading to a mutant histone variant, H3.3-G34W, as a sole recurrent genetic alteration. We show that in patient-derived stromal cells H3.3-G34W is incorporated into the chromatin and associates with massive epigenetic alterations on the DNA methylation, chromatin accessibility and histone modification level, that can be partially recapitulated in an orthogonal cell line system by the introduction of H3.3-G34W. These epigenetic alterations affect mainly heterochromatic and bivalent regions and provide possible explanations for the genomic instability, as well as the osteolytic phenotype of GCTB. The mutation occurs in differentiating mesenchymal stem cells and associates with an impaired osteogenic differentiation. We propose that the observed epigenetic alterations reflect distinct differentiation stages of H3.3 WT and H3.3 MUT stromal cells and add to H3.3-G34W-associated changes