Cdx4 and Menin Co-Regulate Hoxa9 Expression in Hematopoietic Cells

BACKGROUND: Transcription factor Cdx4 and transcriptional coregulator menin are essential for Hoxa9 expression and normal hematopoiesis. However, the precise mechanism underlying Hoxa9 regulation is not clear. METHODS AND FINDINGS: Here, we show that the expression level of Hoxa9 is correlated with the location of increased trimethylated histone 3 lysine 4 (H3K4M3). The active and repressive histone modifications co-exist along the Hoxa9 regulatory region. We further demonstrate that both Cdx4 and menin bind to the same regulatory region at the Hoxa9 locus in vivo, and co-activate the reporter gene driven by the Hoxa9 cis-elements that contain Cdx4 binding sites. Ablation of menin abrogates Cdx4 access to the chromatin target and significantly reduces both active and repressive histone H3 modifications in the Hoxa9 locus. CONCLUSION: These results suggest a functional link among Cdx4, menin and histone modifications in Hoxa9 regulation in hematopoietic cells

NF-Y Dependent Epigenetic Modifications Discriminate between Proliferating and Postmitotic Tissue

The regulation of gene transcription requires posttranslational modifications of histones that, in concert with chromatin remodeling factors, shape the structure of chromatin. It is currently under intense investigation how this structure is modulated, in particular in the context of proliferation and differentiation. Compelling evidence suggests that the transcription factor NF-Y acts as a master regulator of cell cycle progression, activating the transcription of many cell cycle regulatory genes. However, the underlying molecular mechanisms are not yet completely understood. Here we show that NF-Y exerts its effect on transcription through the modulation of the histone “code”. NF-Y colocalizes with nascent RNA, while RNA polymerase II is I phosphorylated on serine 2 of the YSPTSPS repeats within its carboxyterminal domain and histones are carrying modifications that represent activation signals of gene expression (H3K9ac and PAN-H4ac). Comparing postmitotic muscle tissue from normal mice and proliferating muscles from mdx mice, we demonstrate by chromatin immunoprecipitation (ChIP) that NF-Y DNA binding activity correlates with the accumulation of acetylated histones H3 and H4 on promoters of key cell cycle regulatory genes, and with their active transcription. Accordingly, p300 is recruited onto the chromatin of NF-Y target genes in a NF-Y-dependent manner, as demonstrated by Re-ChIP. Conversely, the loss of NF-Y binding correlates with a decrease of acetylated histones, the recruitment of HDAC1, and a repressed heterochromatic state with enrichment of histones carrying modifications known to mediate silencing of gene expression (H3K9me3, H3K27me2 and H4K20me3). As a consequence, NF-Y target genes are downregulated in this context. In conclusion, our data indicate a role of NF-Y in modulating the structure and transcriptional competence of chromatin in vivo and support a model in which NF-Y-dependent histone “code” changes contribute to the proper discrimination between proliferating and postmitotic cells in vivo and in vitro

Skill or Will? Effects of Teacher Knowledge and Motivation on the Quality of Technology-Enhanced Lesson Plans

In an expertise study with 94 mathematics teachers varying in their teaching experience (i.e., pre-service, induction, in-service teachers), we examined effects of teachers’ professional knowledge and motivational beliefs on their ability to effectively integrate educational technology into their lesson plans. We assessed teachers’ professional knowledge (i.e., content knowledge, pedagogical content knowledge, technological knowledge), and their motivational beliefs (i.e., self-efficacy, utility-value). Additionally, teachers developed a worked-out lesson plan for introducing the Pythagorean theorem to secondary students. They were explicitly instructed to refer to the integration of technology in their lesson plans. Experienced teachers considered more cognitively activating tasks in their lesson plans and exploited the potential of technology more than inexperienced teachers. Mediation analyses revealed that this effect was explained by teachers’ perceived utility-value of educational technology but not by their professional knowledge. These findings suggest that teachers’ motivational beliefs play a decisive role for effectively integrating technology in mathematics instruction

Carrier free 10Be/9Be measurements with low-energy AMS: Determination of sedimentation rates in the Arctic Ocean

Using the TANDY AMS facility (600 kV) at ETH Zurich the seawater-derived (authigenic) Be-10/Be-9 ratio of marine sediment samples is measured without the addition of Be-9 carrier. This novel method reduces systematic uncertainties because the Be-10/Be-9 ratio of a sample is determined in only one (AMS) measurement. A challenge of carrier-free AMS is to avoid any contamination of the sample with Be-9 during the chemical preparation. Further, the leaching procedure has to be reproducible and ideally should attack the authigenic Be of the sediments only, leaving the detrital Be untouched. The low amount of stable Be-9 in the unspiked samples causes low currents during the AMS measurement. This requires a good stability and sensitivity of the AMS setup. Our first results show that the new preparation method is reliable and that background from stable Be-9 is avoided. For a comparison study, sediment samples from two cores located in the Arctic Ocean (HLY0503-09JPC, HLY0503-14JPC) were used. The authigenic Be-10/Be-9 ratio of these samples had been determined previously applying the conventional method where Be-10 and Be-9 concentrations are measured separately by AMS and ICP-MS, respectively. The resulting sedimentation rates are in discrepancy with values derived from biomarkers. To cross check the Be-10/Be-9 based age model two samples from each core were measured again with the new carrier-free method. The carrier-free results show systematically higher authigenic Be-10/Be-9 ratios. The calculated sedimentation rates of about 0.2 cm/kyr, however, are consistent for the carrier free and the conventional method

Selective mineral transport barriers at Cuscuta-host infection sites

The uptake of inorganic nutrients by rootless parasitic plants, which depend on host connections for all nutrient supplies, is largely uncharted. Using X‐ray fluorescence spectroscopy (XRF), we analyzed the element composition of macro‐ and micronutrients at infection sites of the parasitic angiosperm Cuscuta reflexa growing on hosts of the genus Pelargonium. Imaging methods combining XRF with 2‐D or 3‐D (confocal) microscopy show that most of the measured elements are present at similar concentrations in the parasite compared to the host. However, calcium and strontium levels drop pronouncedly at the host/parasite interface, and manganese appears to accumulate in the host tissue surrounding the interface. Chlorine is present in the haustorium at similar levels as in the host tissue but is decreased in the stem of the parasite. Thus, our observations indicate a restricted uptake of calcium, strontium, manganese and chlorine by the parasite. Xylem‐mobile dyes, which can probe for xylem connectivity between host and parasite, provided evidence for an interspecies xylem flow, which in theory would be expected to carry all of the elements indiscriminately. We thus conclude that inorganic nutrient uptake by the parasite Cuscuta is regulated by specific selective barriers whose existence has evaded detection until now