Holocarboxylase synthetase regulates expression of biotin transporters by chromatin remodeling events at the SMVT locus

The sodium-dependent multivitamin transporter (SMVT) is essential for mediating and regulating biotin entry into mammalian cells. In cells, biotin is covalently linked to histones in a reaction catalyzed by holocarboxylase synthetase (HCS); biotinylation of lysine 12-biotinylated histone H4 (K12Bio H4) causes gene silencing. Here, we propose a novel role for HCS in sensing and regulating levels of biotin in eukaryotic cells. We hypothesized that nuclear translocation of HCS increases in response to biotin supplementation; HCS then biotinylates histone H4 at SMVT promoters, silencing biotin transporter genes. Jurkat lymphoma cells were cultured in media containing 0.025, 0.25, or 10 nmol/l biotin. The nuclear translocation of HCS correlated with biotin concentrations in media; the relative enrichment of both HCS and K12Bio H4 at SMVT promoter 1 (but not promoter 2) increased by 91% in cells cultured in medium containing 10 nmol/l biotin compared with 0.25 nmol/l biotin. This increase of K12Bio H4 at the SMVT promoter decreased SMVT expression by up to 86%. Biotin homeostasis by HCS-dependent chromatin remodeling at the SMVT promoter 1 locus was disrupted in HCS knockdown cells, as evidenced by abnormal chromatin structure (K12Bio H4 abundance) and increased SMVT expression. The findings from this study are consistent with the theory that HCS senses biotin, and that biotin regulates its own cellular uptake by participating in HCS-dependent chromatin remodeling events at the SMVT promoter 1 locus in Jurkat cells.Fil: Gralla, Michael. Universidad de Nebraska - Lincoln; Estados UnidosFil: Camporeale, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Universidad de Nebraska - Lincoln; Estados UnidosFil: Zempleni, Janos. Universidad de Nebraska - Lincoln; Estados Unido

Identification of novel pathways of regulation of AHR and HIF-­1 function

Mammalian bHLH-Pas (basic HLH (helix-loop-helix)-PER-ARNT-SIM) proteins belong to the bHLH superfamily of transcription factors. Members of the family have a broad spectrum of functions that among others sense and regulate the cellular response to physiological signals such as low oxygen levels (hypoxia), or environmental signals such as toxins. The bHLH-Pas proteins consist of a signal-induced subunit, for example, aryl hydrocarbon receptor (AhR) or hypoxia inducible factor (HIF). To respond to environmental (AhR) or physiological (HIF) stimuli, the signal-induced subunits dimerize with their ubiquitously expressed, unregulated dimerization partner aryl hydrocarbon receptor nuclear translocator (ARNT), bind to DNA, and induce the activation of a cascade of target genes. At normal oxygen levels (normoxia), proline hydroxylases hydroxylate HIF, and the subsequent interaction of HIF with the von Hippel-Lindau tumor suppressor gene (pVHL) leads to ubiquitination and degradation of HIF. Asparagine hydroxylation by the factor inhibiting HIF (FIH) leads under normoxia to suppressed HIF transactivation. According to the model, proline and asparagine hydroxylation reactions are inhibited under hypoxia. In paper I, we established a mechanism that allows us to explain the molecular switch of the aryl hydrocarbon receptor (AhR) from its transcription factor to its E3 ubiquitin ligase function. We used the breast cancer cell line MCF7 to demonstrate that the availability of ARNT modulates the dual functions of AhR. Upon ARNT availability, the AhR functions as a ligand-induced transcription factor. If, however, other binding partners, such as the repressor of AhR (AhRR), occupy ARNT, the AhR functions as an E3 ubiquitin ligase. In paper II, we revealed significantly elevated FIH expression in skeletal muscle compared to other tissues. We also demonstrated that FIH loss leads to an induced oxidative metabolism, and an increased glycolytic capacity, resulting in elevated oxygen consumption. Loss of FIH further correlates with a decreased metabolic efficiency, an increased oxidative rate and an accelerated HIF-mediated response to hypoxia. In paper III (manuscript), we investigated the role FIH plays in epigenetic regulation, among others to explain the underlying mechanism behind the results presented in paper II. Muscle cells show the highest FIH levels on tissue level and we observed elevated histone 3 lysine 9 dimethyl (H3K9me2) levels in FIH null mice. Further work showed significantly increased methylation levels at the H3K9 dimethyl repression mark at promoter regions of various metabolic HIF target genes in FIH null mouse embryonic fibroblasts, and subsequently correlations of loss of FIH and Jmjd1a dysregulated protein levels. In the following, despite promising preliminary results to date no FIH-Jmjd1a protein-protein interaction could be demonstrated

Stellar Populations of Highly Magnified Lensed Galaxies: Young Starbursts at z~2

We present a comprehensive analysis of the rest-frame UV to near-IR spectral energy distributions and rest-frame optical spectra of four of the brightest gravitationally lensed galaxies in the literature: RCSGA 032727-132609 at z=1.70, MS1512-cB58 at z=2.73, SGAS J152745.1+065219 at z=2.76 and SGAS J122651.3+215220 at z=2.92. This includes new Spitzer imaging for RCSGA0327 as well as new spectra, near-IR imaging and Spitzer imaging for SGAS1527 and SGAS1226. Lensing magnifications of 3-4 magnitudes allow a detailed study of the stellar populations and physical conditions. We compare star formation rates as measured from the SED fit, the H-alpha and [OII] emission lines, and the UV+IR bolometric luminosity where 24 micron photometry is available. The SFR estimate from the SED fit is consistently higher than the other indicators, which suggests that the Calzetti dust extinction law used in the SED fitting is too flat for young star-forming galaxies at z~2. Our analysis finds similar stellar population parameters for all four lensed galaxies: stellar masses 3-7*10^9 M_sun, young ages ~ 100 Myr, little dust content E(B-V)=0.10-0.25, and star formation rates around 20-100 M_sun/yr. Compared to typical values for the galaxy population at z~2, this suggests we are looking at newly formed, starbursting systems that have only recently started the build-up of stellar mass. These results constitute the first detailed, uniform analysis of a sample of the growing number of strongly lensed galaxies known at z~2.Comment: 13 pages, 8 figures, Accepted to Ap

Programming gene expression with combinatorial promoters

Promoters control the expression of genes in response to one or more transcription factors (TFs). The architecture of a promoter is the arrangement and type of binding sites within it. To understand natural genetic circuits and to design promoters for synthetic biology, it is essential to understand the relationship between promoter function and architecture. We constructed a combinatorial library of random promoter architectures. We characterized 288 promoters in Escherichia coli, each containing up to three inputs from four different TFs. The library design allowed for multiple −10 and −35 boxes, and we observed varied promoter strength over five decades. To further analyze the functional repertoire, we defined a representation of promoter function in terms of regulatory range, logic type, and symmetry. Using these results, we identified heuristic rules for programming gene expression with combinatorial promoters

A measurement of the millimetre emission and the Sunyaev–Zel'dovich effect associated with low-frequency radio sources

We present a statistical analysis of the millimetre-wavelength properties of 1.4 GHz-selected sources and a detection of the Sunyaev–Zel'dovich (SZ) effect associated with the haloes that host them. We stack data at 148, 218 and 277 GHz from the Atacama Cosmology Telescope at the positions of a large sample of radio AGN selected at 1.4 GHz. The thermal SZ effect associated with the haloes that host the AGN is detected at the 5σ level through its spectral signature, representing a statistical detection of the SZ effect in some of the lowest mass haloes (average M_(200) ≈ 10^(13) M⊙ h^(−1)_(70) studied to date. The relation between the SZ effect and mass (based on weak lensing measurements of radio galaxies) is consistent with that measured by Planck for local bright galaxies. In the context of galaxy evolution models, this study confirms that galaxies with radio AGN also typically support hot gaseous haloes. Adding Herschel observations allows us to show that the SZ signal is not significantly contaminated by dust emission. Finally, we analyse the contribution of radio sources to the angular power spectrum of the cosmic microwave background

The Atacama Cosmology Telescope: Cross Correlation with Planck maps

We present the temperature power spectrum of the Cosmic Microwave Background obtained by cross-correlating maps from the Atacama Cosmology Telescope (ACT) at 148 and 218 GHz with maps from the Planck satellite at 143 and 217 GHz, in two overlapping regions covering 592 square degrees. We find excellent agreement between the two datasets at both frequencies, quantified using the variance of the residuals between the ACT power spectra and the ACTxPlanck cross-spectra. We use these cross-correlations to calibrate the ACT data at 148 and 218 GHz, to 0.7% and 2% precision respectively. We find no evidence for anisotropy in the calibration parameter. We compare the Planck 353 GHz power spectrum with the measured amplitudes of dust and cosmic infrared background (CIB) of ACT data at 148 and 218 GHz. We also compare planet and point source measurements from the two experiments.Comment: 9 pages, 8 figure

Prediction of Long-Term Restenosis After Carotid Endarterectomy Using Quantitative Magnetic Resonance Angiography.

Background To detect restenosis after carotid endarterectomy (CEA), long-term monitoring is required. However, non-selective follow-up is controversial and can be limited by costs and logistical considerations. Objective To examine the value of immediate perioperative vessel flow measurements after CEA using quantitative magnetic resonance angiography (QMRA) to detect patients at risk of long-term restenosis. Methods A prospective cohort study with long-term sonographic follow-up after CEA for symptomatic internal carotid artery stenosis (ICAs) > 50%. In all patients, vessel flow has been assessed both pre- and postoperatively using QMRA within ±3 days of surgery. Data on QMRA assessment were analyzed to identify patients at risk of restenosis for up to 10 years. Results Restenosis was recorded in 4 of 24 patients (17%) at a median follow-up of 6.8 ± 2.6 years. None of them experienced an ischemic event. Perioperative flow differences were significantly greater in patients without long-term restenosis, both for the ipsilateral ICA (p < 0.001) and MCA (p = 0.03), compared to those with restenosis (p = 0.22 and p = 0.3, respectively). The ICA mean flow ratio (p = 0.05) tended to be more effective than the MCA ratio in predicting restenosis over the long term (p = 0.35). Conclusion Our preliminary findings suggest that QMRA-based mean flow increases after CEA may be predictive of restenosis over the long term. Perioperative QMRA assessment could become an operator-independent screening tool to identify a subgroup of patients at risk for restenosis, in whom long-term monitoring is advised

The Atacama Cosmology Telescope: A Measurement of the Thermal Sunyaev-Zel'dovich Effect Using the Skewness of the CMB Temperature Distribution

We present a detection of the unnormalized skewness induced by the thermal Sunyaev-Zel'dovich (tSZ) effect in filtered Atacama Cosmology Telescope (ACT) 148 GHz cosmic microwave background temperature maps. Contamination due to infrared and radio sources is minimized by template subtraction of resolved sources and by constructing a mask using outlying values in the 218 GHz (tSZ-null) ACT maps. We measure = -31 +- 6 \mu K^3 (measurement error only) or +- 14 \mu K^3 (including cosmic variance error) in the filtered ACT data, a 5-sigma detection. We show that the skewness is a sensitive probe of sigma_8, and use analytic calculations and tSZ simulations to obtain cosmological constraints from this measurement. From this signal alone we infer a value of sigma_8= 0.79 +0.03 -0.03 (68 % C.L.) +0.06 -0.06 (95 % C.L.). Our results demonstrate that measurements of non-Gaussianity can be a useful method for characterizing the tSZ effect and extracting the underlying cosmological information.Comment: 9 pages, 5 figures. Replaced with version accepted by Phys. Rev. D, with improvements to the likelihood function and the IR source treatment; only minor changes in the result

Cosmological Parameters from Pre-Planck CMB Measurements

Recent data from the WMAP, ACT and SPT experiments provide precise measurements of the cosmic microwave background temperature power spectrum over a wide range of angular scales. The combination of these observations is well fit by the standard, spatially flat LCDM cosmological model, constraining six free parameters to within a few percent. The scalar spectral index, n_s = 0.9690 +/- 0.0089, is less than unity at the 3.6 sigma level, consistent with simple models of inflation. The damping tail of the power spectrum at high resolution, combined with the amplitude of gravitational lensing measured by ACT and SPT, constrains the effective number of relativistic species to be N_eff = 3.28 +/- 0.40, in agreement with the standard model's three species of light neutrinos.Comment: 5 pages, 4 figure