TALE Factors Poise Promoters for Activation by Hox Proteins

SummaryHox proteins form complexes with TALE cofactors from the Pbx and Prep/Meis families to control transcription, but it remains unclear how Hox:TALE complexes function. Examining a Hoxb1b:TALE complex that regulates zebrafish hoxb1a transcription, we find maternally deposited TALE proteins at the hoxb1a promoter already during blastula stages. These TALE factors recruit histone-modifying enzymes to promote an active chromatin profile at the hoxb1a promoter and also recruit RNA polymerase II (RNAPII) and P-TEFb. However, in the presence of TALE factors, RNAPII remains phosphorylated on serine 5 and hoxb1a transcription is inefficient. By gastrula stages, Hoxb1b binds together with TALE factors to the hoxb1a promoter. This triggers P-TEFb-mediated transitioning of RNAPII to the serine 2-phosphorylated form and efficient hoxb1a transcription. We conclude that TALE factors access promoters during early embryogenesis to poise them for activation but that Hox proteins are required to trigger efficient transcription

Determination of structural parameters characterizing thin films by optical methods: A comparison between scanning angle reflectometry and optical waveguide lightmode spectroscopy

International audienceWe present a comparative study of the structural parameters characterizing thin macromolecular adsorbed films that are obtained from two optical techniques: optical waveguide lightmode spectroscopy ͑OWLS͒ and scanning angle reflectometry ͑SAR͒. We use polyelectrolyte multilayers and polyelectrolyte multilayers/protein films to perform this study. The comparison between the information obtained with the two methods is possible because the buildup of the polyelectrolyte multilayers is known to become substrate independent after the deposition of the first few polyelectrolyte layers. The analysis of the optical data requires usually to postulate a refractive index profile for the interface. Two profiles have been used: the homogeneous and isotropic monolayer and the bilayer profiles. When the refractive index profile of an adsorbed film is well approximated by a homogeneous and isotropic monolayer, as shown by using an analysis of the deposited films in terms of optical invariants, the two optical techniques lead to similar values for the film thickness and the optical mass. The situation is more complex in the case of the multilayers/protein films for which the calculated parameters can strongly depend upon the refractive index profile that is postulated to analyze the optical data. Whereas the optical mass and, to a lesser extent, the thickness seem fairly model independent for OWLS, they appear to be extremely sensitive to the model for SAR. For proteins deposited on top of the polyelectrolyte film, optical mass and protein thickness were found to be comparable when determined by OWLS and by SAR using the bilayer model. The data analysis of the SAR curves with the monolayer model leads to much larger and even physically unreasonable film thicknesses and optical masses. This was particularly noticeable for proteins having a large size ͑human serum albumin and fibrinogen͒, whereas both models lead to similar results for small sized proteins. By means of the different refractive index profiles, we show that great care must be taken in the physicochemical interpretation of the structural parameters determined by these optical techniques

TALE and NF-Y co-occupancy marks enhancers of developmental control genes during zygotic genome activation in zebrafish [preprint]

Animal embryogenesis is initiated by maternal factors, but zygotic genome activation (ZGA) shifts control to the embryo at early blastula stages. ZGA is thought to be mediated by specialized maternally deposited transcription factors (TFs), but here we demonstrate that NF-Y and TALE – TFs with known later roles in embryogenesis – co-occupy unique genomic elements at zebrafish ZGA. We show that these elements are selectively associated with early-expressed genes involved in transcriptional regulation and possess enhancer activity in vivo. In contrast, we find that elements individually occupied by either NF-Y or TALE are associated with genes acting later in development – such that NF-Y controls a cilia gene expression program while TALE TFs control expression of hox genes. We conclude that NF-Y and TALE have a shared role at ZGA, but separate roles later during development, demonstrating that combinations of known TFs can regulate subsets of key developmental genes at vertebrate ZGA

Polyelectrolyte Multilayering on a Charged Planar Surface

The adsorption of highly \textit{oppositely} charged flexible polyelectrolytes (PEs) on a charged planar substrate is investigated by means of Monte Carlo (MC) simulations. We study in detail the equilibrium structure of the first few PE layers. The influence of the chain length and of a (extra) non-electrostatic short range attraction between the polycations and the negatively charged substrate is considered. We show that the stability as well as the microstructure of the PE layers are especially sensitive to the strength of this latter interaction. Qualitative agreement is reached with some recent experiments.Comment: 28 pages; 11 (main) Figs - Revtex4 - Higher resolution Figs can be obtained upon request. To appear in Macromolecule

A Cryogenic High-Reynolds Turbulence Experiment at CERN

The potential of cryogenic helium flows for studying high-Reynolds number turbulence in the laboratory has been recognised for a long time and implemented in several small-scale hydrodynamic experiments. With its large superconducting particle accelerators and detector magnets, CERN, the European Laboratory for Particle Physics, has become a major world center in helium cryogenics, with several large helium refrigerators having capacities up to 18 kW @ 4.5 K. Combining a small fraction of these resources with the expertise of three laboratories at the forefront of turbulence research, has led to the design, swift implementation, and successful operation of GReC (Grands Reynolds Cryogéniques) a large axisymmetric turbulent-jet experiment. With flow-rates up to 260 g/s of gaseous helium at ~ 5 K and atmospheric pressure, Reynolds numbers up to 107 have been achieved in a 4.6 m high, 1.4 m diameter cryostat. This paper presents the results of the first runs and describes the experimental set-up comprehensively equipped with "hot" wire micro-anemometers, acoustic scattering vorticity measurements and a large-bandwidth data acquisition system

Probing helium interfaces with light scattering : from fluid mechanics to statistical physics

We have investigated the formation of helium droplets in two physical situations. In the first one, droplets are atomised from superfluid or normal liquid by a fast helium vapour flow. In the second, droplets of normal liquid are formed inside porous glasses during the process of helium condensation. The context, aims, and results of these experiments are reviewed, with focus on the specificity of light scattering by helium. In particular, we discuss how, for different reasons, the closeness to unity of the index of refraction of helium allows in both cases to minimise the problem of multiple scattering and obtain results which it would not be possible to get using other fluids.Comment: 21 page

« Half dicht, half prose gheordineert » : vers et prose de moyen français en moyen néerlandais

In both French-speaking and Dutch-speaking literary cultures of the late Middle Ages, competition between poets produced a collective poetic expertise. To what extent, then, can such competition be identified across the two cultures, in translations of verse or prosimetrum compositions from Middle French into Middle Dutch? An examination of the Dutch translations reveals that verse is both a means to knowledge and an object of knowledge, in the target culture as well as the source culture. The diversity of translations shows that verse is not only a system that translators attempt to master, but also a formal supplement in ways that are unavailable to prose

Grafted ionomer complexes and their effect on protein adsorption on silica and polysulfone surfaces

We have studied the formation and the stability of ionomer complexes from grafted copolymers (GICs) in solution and the influence of GIC coatings on the adsorption of the proteins β-lactoglobulin (β-lac), bovine serum albumin (BSA), and lysozyme (Lsz) on silica and polysulfone. The GICs consist of the grafted copolymer PAA28-co-PAPEO22 {poly(acrylic acid)-co-poly[acrylate methoxy poly(ethylene oxide)]} with negatively charged AA and neutral APEO groups, and the positively charged homopolymers: P2MVPI43 [poly(N-methyl 2-vinyl pyridinium iodide)] and PAH∙HCl160 [poly(allylamine hydrochloride)]. In solution, these aggregates are characterized by means of dynamic and static light scattering. They appear to be assemblies with hydrodynamic radii of 8 nm (GIC-PAPEO22/P2MVPI43) and 22 nm (GIC-PAPEO22/PAH∙HCl160), respectively. The GICs partly disintegrate in solution at salt concentrations above 10 mM NaCl. Adsorption of GICs and proteins has been studied with fixed angle optical reflectometry at salt concentrations ranging from 1 to 50 mM NaCl. Adsorption of GICs results in high density PEO side chains on the surface. Higher densities were obtained for GICs consisting of PAH∙HCl160 (1.6 ÷ 1.9 chains/nm2) than of P2MVPI43 (0.6 ÷ 1.5 chains/nm2). Both GIC coatings strongly suppress adsorption of all proteins on silica (>90%); however, reduction of protein adsorption on polysulfone depends on the composition of the coating and the type of protein. We observed a moderate reduction of β-lac and Lsz adsorption (>60%). Adsorption of BSA on the GIC-PAPEO22/P2MVPI43 coating is moderately reduced, but on the GIC-PAPEO22/PAH∙HCl160 coating it is enhanced

Hox regulation of transcription: more complex(es)

Hox genes encode transcription factors with important roles during embryogenesis and tissue differentiation. Genetic analyses initially demonstrated that interfering with Hox genes has profound effects on the specification of cell identity, suggesting that Hox proteins regulate very specific sets of target genes. However, subsequent biochemical analyses revealed that Hox proteins bind DNA with relatively low affinity and specificity. Furthermore, it became clear that a given Hox protein could activate or repress transcription, depending on the context. A resolution to these paradoxes presented itself with the discovery that Hox proteins do not function in isolation, but interact with other factors in complexes. The first such cofactors were members of the Extradenticle/Pbx and Homothorax/Meis/Prep families. However, the list of Hox-interacting proteins has continued to grow, suggesting that Hox complexes contain many more components than initially thought. Additionally, the activities of the various components and the exact mechanisms whereby they modulate the activity of the complex remain puzzling. Here, we review the various proteins known to participate in Hox complexes and discuss their likely functions. We also consider that Hox complexes of different compositions may have different activities and discuss mechanisms whereby Hox complexes may be switched between active and inactive states