Epigenetic Regulation of a Murine Retrotransposon by a Dual Histone Modification Mark

Large fractions of eukaryotic genomes contain repetitive sequences of which the vast majority is derived from transposable elements (TEs). In order to inactivate those potentially harmful elements, host organisms silence TEs via methylation of transposon DNA and packaging into chromatin associated with repressive histone marks. The contribution of individual histone modifications in this process is not completely resolved. Therefore, we aimed to define the role of reversible histone acetylation, a modification commonly associated with transcriptional activity, in transcriptional regulation of murine TEs. We surveyed histone acetylation patterns and expression levels of ten different murine TEs in mouse fibroblasts with altered histone acetylation levels, which was achieved via chemical HDAC inhibition with trichostatin A (TSA), or genetic inactivation of the major deacetylase HDAC1. We found that one LTR retrotransposon family encompassing virus-like 30S elements (VL30) showed significant histone H3 hyperacetylation and strong transcriptional activation in response to TSA treatment. Analysis of VL30 transcripts revealed that increased VL30 transcription is due to enhanced expression of a limited number of genomic elements, with one locus being particularly responsive to HDAC inhibition. Importantly, transcriptional induction of VL30 was entirely dependent on the activation of MAP kinase pathways, resulting in serine 10 phosphorylation at histone H3. Stimulation of MAP kinase cascades together with HDAC inhibition led to simultaneous phosphorylation and acetylation (phosphoacetylation) of histone H3 at the VL30 regulatory region. The presence of the phosphoacetylation mark at VL30 LTRs was linked with full transcriptional activation of the mobile element. Our data indicate that the activity of different TEs is controlled by distinct chromatin modifications. We show that activation of a specific mobile element is linked to a dual epigenetic mark and propose a model whereby phosphoacetylation of histone H3 is crucial for full transcriptional activation of VL30 elements

Effect of solids retention time on Microthrix parvicella growth

The objective of this study was to evaluate the effect of solids retention time (SRT) on M. parvicella growth and to calculate growth kinetic parameters of this filamentous species. Bench-scale continuous-flow experiments showed that M. parvicella growth can be significantly suppressed at an SRT of lower than 5.7 d for temperatures of between 14 and 18oC. According to the continuous-flow experiments the maximum sludge age for the avoidance of filamentous foaming problems caused by M. parvicella is 6 d for temperatures lower than 18oC. At this sludge age M. parvicella loses its hydrophobicity and therefore its foaming potential. However, even lower SRTs are required in order to achieve a significant suppression of its growth. At SRT values of less than 5.7 d M. parvicella initially forms a shorter filament (< 150 μm) with clear spaces inside filaments and variable Gram stain reaction and eventually is eliminated from activated sludge biocoenosis. According to kinetic studies presented in this paper, M. parvicella is a slow growing bacterium with a low maximum specific growth rate of 0.67 1/d and 0.53 1/d under aerobic and anoxic conditions respectively. Maintenance energy requirements of M. parvicella were found to be significantly lower than the maintenance energy of floc forming micro-organisms as well as other filamentous species, thus providing the micro-organism with a significant advantage under starvation conditions prevailing at the majority of the extended aeration activated sludge systems