Xuecong Wang

Linc-YY1 promotes myogenic differentiation and muscle regeneration through an interaction with the transcription factor YY1

Little is known how lincRNAs are involved in skeletal myogenesis. Here we describe the discovery of Linc-YY1 from the promoter of the transcription factor (TF) Yin Yang 1 (YY1) gene. We demonstrate that Linc-YY1 is dynamically regulated during myogenesis in vitro and in vivo. Gain or loss of function of Linc-YY1 in C2C12 myoblasts or muscle satellite cells alters myogenic differentiation and in injured muscles has an impact on the course of regeneration. Linc-YY1 interacts with YY1 through its middle domain, to evict YY1/Polycomb repressive complex (PRC2) from target promoters, thus activating the gene expression in trans. In addition, Linc-YY1 also regulates PRC2-independent function of YY1. Finally, we identify a human Linc-YY1 orthologue with conserved function and show that many human and mouse TF genes are associated with lincRNAs that may modulate their activity. Altogether, we show that Linc-YY1 regulates skeletal myogenesis and uncover a previously unappreciated mechanism of gene regulation by lincRNA.

Structure and decoy-mediated inhibition of the SOX18/Prox1-DNA interaction

The transcription factor (TF) SOX18 drives lymphatic vessel development in both embryogenesis and tumour-induced neo-lymphangiogenesis. Genetic disruption of Sox18 in a mouse model protects from tumour metastasis and established the SOX18 protein as a molecular target. Here, we report the crystal structure of the SOX18 DNA binding high-mobility group (HMG) box bound to a DNA element regulating Prox1 transcription. The crystals diffracted to 1.75Å presenting the highest resolution structure of a SOX/DNA complex presently available revealing water structure, structural adjustments at the DNA contact interface and non-canonical conformations of the DNA backbone. To explore alternatives to challenging small molecule approaches for targeting the DNA-binding activity of SOX18, we designed a set of five decoys based on modified Prox1-DNA. Four decoys potently inhibited DNA binding of SOX18 in vitro and did not interact with non-SOX TFs. Serum stability, nuclease resistance and thermal denaturation assays demonstrated that a decoy circularized with a hexaethylene glycol linker and terminal phosphorothioate modifications is most stable. This SOX decoy also interfered with the expression of a luciferase reporter under control of a SOX18-dependent VCAM1 promoter in COS7 cells. Collectively, we propose SOX decoys as potential strategy for inhibiting SOX18 activity to disrupt tumour-induced neo-lymphangiogenesis.

OCT4: A penetrant pluripotency inducer

Native OCT4 protein has the intrinsic ability of crossing cellular membranes to enter cells. This finding could revive efforts to induce pluripotency with proteins replacing nucleic acid-based approaches, and raises the intriguing question as to whether OCT4 can act non-cell-autonomously.

A HIGH FREQUENCY THERMOACOUSTICALLY‐DRIVEN PULSE TUBE CRYOCOOLER WITH COAXIAL RESONATOR

High frequency thermoacoustically‐driven pulse tube cryocoolers are quite promising due to their compact size and high reliability, which can find applications in space use. With continuous effort, a lowest cold head temperature of 68.3 K has been obtained on a 300 Hz pulse tube cryocooler driven by a standing‐wave thermoacoustic heat engine with 4.0 MPa helium gas and 750 W heat input. To further reduce the size of the system, a coaxial resonator was designed and the two sub‐systems, i.e., the pulse tube cryocooler and the standing‐wave thermoacoustic heat engine were properly coupled through an acoustic amplifier tube, which leads to a system axial length of only about 0.7 m. The performance of the system with the coaxial resonator was tested, and shows moderate degradation compared to that with the in‐line resonator, which might be attributed to the large flow loss of the 180 degree corner.

DNA-mediated dimerization on a compact sequence signature controls enhancer engagement and regulation by FOXA1

Abstract FOXA1 is a transcription factor capable to bind silenced chromatin to direct context-dependent cell fate conversion. Here, we demonstrate that a compact palindromic DNA element (termed ‘DIV’ for its diverging half-sites) induces the homodimerization of FOXA1 with strongly positive cooperativity. Alternative structural models are consistent with either an indirect DNA-mediated cooperativity or a direct protein-protein interaction. The cooperative homodimer formation is strictly constrained by precise half-site spacing. Re-analysis of chromatin immunoprecipitation sequencing data indicates that the DIV is effectively targeted by FOXA1 in the context of chromatin. Reporter assays show that FOXA1-dependent transcriptional activity declines when homodimeric binding is disrupted. In response to phosphatidylinositol-3 kinase inhibition DIV sites pre-bound by FOXA1 such as at the PVT1/MYC locus exhibit a strong increase in accessibility suggesting a role of the DIV configuration in the chromatin closed-open dynamics. Moreover, several disease-associated single nucleotide polymorphisms map to DIV elements and show allelic differences in FOXA1 homodimerization, reporter gene expression and are annotated as quantitative trait loci. This includes the rs541455835 variant at the MAPT locus encoding the Tau protein associated with Parkinsons disease. Collectively, the DIV guides chromatin engagement and regulation by FOXA1 and its perturbation could be linked to disease etiologies.

Identification of a Wells–Dawson polyoxometalate-based AP-2γ inhibitor with pro-apoptotic activity

AP-2 gamma (AP-2γ) is a transcription factor that plays pivotal roles in breast cancer biology. To search for small molecule inhibitors of AP-2γ, we performed a high-throughput fluorescence anisotropy screen and identified a polyoxometalate compound with Wells-Dawson structure K6[P2Mo18O62] (Dawson-POM) that blocks the DNA-binding activity of AP-2γ. We showed that this blocking activity is due to the direct binding of Dawson-POM to AP-2γ. We also provided evidence to show that Dawson-POM decreases AP-2γ-dependent transcription similar to silencing the gene. Finally, we demonstrated that Dawson-POM contains anti-proliferative and pro-apoptotic effects in breast cancer cells. In summary, we identified the first small molecule inhibitor of AP-2γ and showed Dawson-POM-mediated inhibition of AP-2γ as a potential avenue for cancer therapy.

Numerical study of a cryogen-free vuilleumier type pulse tube cryocooler operating below 10 K

This paper presents a numerical investigation on a Vuilleumier (VM) type pulse tube cooler. Different from previous systems that use liquid nitrogen, Stirling type pre-coolers are used to provide the cooling power for the thermal compressor, which leads to a convenient cryogen-free system and offers the flexibility of changing working temperature range of the thermal compressor to obtain an optimum efficiency. Firstly, main component dimensions were optimized with lowest no-load temperature as the target. Then the dependence of system performance on average pressure, frequency, displacer displacement amplitude and thermal compressor pre-cooling temperature were studied. Finally, the effect of pre-cooling temperature on overall cooling efficiency at 5 K was studied. A highest relative Carnot efficiency of 0.82 % was predicted with an average pressure of 2.5 MPa, a frequency of 3 Hz, a displacer displacement amplitude of 6.5 mm, ambient end temperature 300 K and pre-cooling temperature 65 K, respectively.