Suppression of Gluconeogenic Gene Expression by LSD1-Mediated Histone Demethylation

Aberrant gluconeogenic gene expression is associated with diabetes, glycogen storage disease, and liver cancer. However, little is known how these genes are regulated at the chromatin level. In this study, we investigated in HepG2 cells whether histone demethylation is a potential mechanism. We found that knockdown or pharmacological inhibition of histone demethylase LSD1 causes remarkable transcription activation of two gluconeogenic genes, FBP1 and G6Pase, and consequently leads to increased de novo glucose synthesis and decreased intracellular glycogen content. Mechanistically, LSD1 occupies the promoters of FBP1 and G6Pase, and modulates their H3K4 dimethylation levels. Thus, our work identifies an epigenetic pathway directly governing gluconeogenic gene expression, which might have important implications in metabolic physiology and diseases

The Histone Demethylase Jhdm1a Regulates Hepatic Gluconeogenesis

Hepatic gluconeogenesis is required for maintaining blood glucose homeostasis; yet, in diabetes mellitus, this process is unrestrained and is a major contributor to fasting hyperglycemia. To date, the impacts of chromatin modifying enzymes and chromatin landscape on gluconeogenesis are poorly understood. Through catalyzing the removal of methyl groups from specific lysine residues in the histone tail, histone demethylases modulate chromatin structure and, hence, gene expression. Here we perform an RNA interference screen against the known histone demethylases and identify a histone H3 lysine 36 (H3K36) demethylase, Jhdm1a, as a key negative regulator of gluconeogenic gene expression. In vivo, silencing of Jhdm1a promotes liver glucose synthesis, while its exogenous expression reduces blood glucose level. Importantly, the regulation of gluconeogenesis by Jhdm1a requires its demethylation activity. Mechanistically, we find that Jhdm1a regulates the expression of a major gluconeogenic regulator, C/EBPα. This is achieved, at least in part, by its USF1-dependent association with the C/EBPα promoter and its subsequent demethylation of dimethylated H3K36 on the C/EBPα locus. Our work provides compelling evidence that links histone demethylation to transcriptional regulation of gluconeogenesis and has important implications for the treatment of diabetes

New Superhard Carbon Phases Between Graphite and Diamond

Two new carbon allotropes (H-carbon and S-carbon) are proposed, as possible candidates for the intermediate superhard phases between graphite and diamond obtained in the process of cold compressing graphite, based on the results of first-principles calculations. Both H-carbon and S-carbon are more stable than previously proposed M-carbon and W-carbon and their bulk modulus are comparable to that of diamond. H-carbon is an indirect-band-gap semiconductor with a gap of 4.459 eV and S-carbon is a direct-band-gap semiconductor with a gap of 4.343 eV. The transition pressure from cold compressing graphite is 10.08 GPa and 5.93 Gpa for H-carbon and S-carbon, respectively, which is in consistent with the recent experimental report.Comment: 5pages,4figures,submitted to Phys.Rev.Lett on 18Jan12, transfer to Phys.Rev.B on 25Mar12; Solid State Communications(2012), http://dx.doi.org/10.1016/j.ssc.2012.05.02

Gate-Level Information Flow Tracking for Security Lattices

High-assurance systems found in safety-critical infrastructures are facing steadily increasing cyber threats. These critical systems require rigorous guarantees in information flow security to prevent confidential information from leaking to an unclassified domain and the root of trust from being violated by an untrusted party. To enforce bit-tight information flow control, gate-level information flow tracking (GLIFT) has recently been proposed to precisely measure and manage all digital information flows in the underlying hardware, including implicit flows through hardware-specific timing channels. However, existing work in this realm either restricts to two-level security labels or essentially targets two-input primitive gates and several simple multilevel security lattices. This article provides a general way to expand the GLIFT method for multilevel security. Specifically, it formalizes tracking logic for an arbitrary Boolean gate under finite security lattices, presents a precise tracking logic generation method for eliminating false positives in GLIFT logic created in a constructive manner, and illustrates application scenarios of GLIFT for enforcing multilevel information flow security. Experimental results show various trade-offs in precision and performance of GLIFT logic created using different methods. It also reveals the area and performance overheads that should be expected when expanding GLIFT for multilevel security

Cloning, Expression and Characterization of a Novel Fructosyltransferase from Aspergillus oryzae ZZ-01 for the Synthesis of Sucrose 6-Acetate

A 1521 bp gene encoding for a novel fructosyltransferase from Aspergillus oryzae ZZ-01 (AoFT) has been amplified by RACE and TAIL PCR, and functionally overexpressed in Escherichia coli BL 21-CodonPlus (DE3)-RIL. The recombinant A. oryzae ZZ-01 fructosyltransferases (r-AoFT) was purified to homogeneity after Ni-NTA affinity and Superdex-200 gel filtration chromatography. SDS-PAGE analysis of the purified r-AoFT revealed a single protein band with an apparent molecular mass of 60.0 kDa. The r-AoFT enzyme exhibited its optimal activity at 55 °C and pH 5.5, and maintained about 63% of its activity even after 60 min of treatment at 60 °C. The addition of Mg2+ led to an increase in the activity of r-AoFT, whereas Zn2+, Cu2+ and Ni2+ led to a reduction in its activity. Six site-directed mutants of r-AoFT (D39A, D164A, E216A, N38L, S99A and Y282A) were constructed and characterized biochemically. The N38L, S99A and Y282A mutants had lower Km and higher Vmax values than the wild-type enzyme, highlighting their higher binding affinity for the substrates. These results therefore suggest that r-AoFT could be used for the enzymatic synthesis of Suc6A from sucrose and glucose 6-acetate

Ni-Catalyzed Deamination Cross-Electrophile Coupling of Katritzky Salts with Halides via C–N Bond Activation

This work describes the first Ni-catalyzed cross-electrophile coupling of alkylpyridinium salts, derived from the corresponding amines, with aryl iodide, bromoalkyne or bromoalkyl coupling partners. C(sp)-C(sp3), C(sp2)-C(sp3) and C(sp3)-C(sp3) bond formation was achieved to afford a variety of synthetically useful arenes, alkynes and alkanes in good yields from2-33. The advantages of the methodology are showcased in the two-step synthesis of the key lactonic moiety of (+)-Compactin and (+)-Mevinolin from commercially available starting materials. A one-pot procedure without isolation of alkylpyridinium tetrafluoroborate salt was also demonstrated to be successful. This work represents a new strategy for the cross-coupling reaction of two electrophiles, and also provides a complementary and highly valuable vista for the current methodologies of alkyl arene/alkyne/alkane synthesis

Regulation of H3K4me2 at FBP1 and G6Pase promoters by LSD1.

<p>HepG2 cells were infected with LSD1 knockdown lentiviruses (<b>A</b>) or treated with TCP (1 mM) (<b>B</b>), and H3K4me2 levels were examined at FBP1, G6Pase and GAPDH promoters. All data were shown from one representative of two independent experiments with similar results.</p

Gene full names and primersequences used in this study.

<p>Gene full names and primersequences used in this study.</p

LSD1 regulates glucose production in HepG2 cells.

<p>Assays were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066294#s4" target="_blank">Materials and Methods</a>. (<b>A, B</b>) gluconeogenesis in LSD1 knockdown cells (<b>A</b>) or in cells pre-treated with TCP for 16 hr (<b>B</b>), n = 4. (<b>C</b>) glycogen content in LSD1 knockdown cells, n = 3. *, P<0.05; **, P<0.01.</p