An Unreasonable Universality of the Thermophoretic Velocity.

Thermophoresis is the migration of dispersed molecules or particles in an inhomogeneous temperature field. It has been associated with various nonequilibrium phenomena ranging from stratified oil reservoirs to prebiotic evolution and the origin of life. The thermophoretic velocity is difficult to predict and appears almost random. We show that, in the case of strongly asymmetric mixtures with high molecular mass ratios of the solute to the solvent, it unexpectedly assumes a universal value once the trivial influence of the viscosity has been factored out. This asymptotic behavior is surprisingly universal and a general property of many highly asymmetric molecular mixtures ranging from organic molecules in n-alkanes to dilute solutions of high polymers. A quantitative explanation is provided on the basis of the asymmetric limit of the pseudoisotopic Soret effect.

Thermophobicity of liquids: heats of transport in mixtures as pure component properties--the case of arbitrary concentration.

We have measured Soret coefficients of a large number of binary mixtures of 23 different organic solvents. The present analysis is based on 77 equimolar mixtures and strongly supports the thermophobicity concept previously developed for the heats of transport of originally 10 different substances [S. Hartmann, G. Wittko, W. Köhler, K. I. Morozov, K. Albers, and G. Sadowski, Phys. Rev. Lett. 109, 065901 (2012)]. Among the investigated compounds, cis-decalin is the most thermophobic, hexane the most thermophilic one. In addition to the equimolar mixtures, we have also analyzed the composition dependence of the Soret coefficients and the heats of transport for 22 selected binary mixtures. Both the interpretation of the heats of transport in equimolar mixtures as pure component thermophobicities and the composition dependence of the Soret coefficient can be understood on the basis of the thermodiffusion theory developed by Morozov [Phys. Rev. E 79, 031204 (2009)], according to which the composition dependence is determined by the excess volume of mixing.

Phenotypic suppression of empty spiracles is prevented by buttonhead.

Unlike the trunk segments, the anterior head segments of Drosophila are formed in the absence of pair-rule and HOX-cluster gene expression, by the activities of the gap-like genes orthodenticle (otd), empty spiracles (ems) and buttonhead (btd). The products of these genes are transcription factors, but only EMS has a HOX-like homeodomain. Indeed, ems can confer identity to trunk segments when other HOX-cluster gene activities are absent. In trunk segments of wild-type embryos, however, ems activity is prevented by phenotypic suppression, in which more posterior HOX-cluster genes inactivate the more anterior without affecting transcription or translation. ems is suppressed by all other Hox-cluster genes and so is placed at the bottom of their hierarchy. Here we show that misexpression of EMS in the head transforms segment identity in a btd-dependent manner, that misexpression of BTD in the trunk causes ems-dependent structures to develop, and that EMS and BTD interact in vitro. The data indicate that this interaction may allow ems to escape from the bottom of the HOX-cluster gene hierarchy and cause a dominant switch of homeotic prevalence in the anterior-posterior direction.

Common and diverged functions of the Drosophila gene pair D-Sp1 and buttonhead.

The Drosophila gene buttonhead (btd) is required for the formation of the mandibular, the intercalary and the antennal head segments of the embryo. The btd protein (BTD) is functionally and structurally related to the human C(2)H(2) zinc finger transcription factor Sp1. A second Sp1-like Drosophila gene, termed Drosophila Sp1 (D-Sp1), had been identified on the basis of a partial sequence showing that the gene encodes a characteristic zinc finger domain, composed of three finger motifs similar to both Sp1 and btd. D-Sp1 is located in the same cytological location as btd in chromosome band 9A on the X-chromosome. It had been proposed that D-Sp1 and btd are likely to act as a gene pair and function in a at least partially redundant manner. Here we report the molecular analysis of D-Sp1 and its expression pattern during embryonic and larval development. We show that D-Sp1 acts as a transcriptional regulator. Lack-of-function analysis combined with rescue and gain-of-function studies indicates that btd and D-Sp1 play essential and redundant roles for mechanosensory organ development. However, D-Sp1 lacks the specific features of BTD required for embryonic intercalary and antennal segment formation.

Drosophila head segmentation factor buttonhead interacts with the same TATA box-binding protein-associated factors and in vivo DNA targets as human Sp1 but executes a different biological program.

The Drosophila gene buttonhead (btd) is required for the establishment of three embryonic head segments. It encodes a zinc-finger-type transcription factor expressed in the corresponding head segment anlagen in the blastoderm stage embryo. The DNA-binding properties of the btd protein (BTD) are indistinguishable from the human transcription factor Sp1. Furthermore, BTD and Sp1 are capable of activating transcription in transfected cultured cells through interaction with the same DNA target sites. Herein we show that BTD and Sp1 functionally interact with the same TATA box-binding protein-associated factors and support in vitro transcription activation through these contacts. Transgene expression of BTD results in the rescue of the head segments that fail to develop in btd mutant embryos, whereas Sp1 or Sp1 containing the zinc finger region of BTD rescues mandibular segment development. The results suggest that BTD contains functional domains other than an equivalent DNA-binding region and interaction sites of the TATA box-binding protein-associated factors, which are necessary to establish head segments that fail to develop in response to Sp1.

Molecular analysis of the interaction between the Bacillus subtilis trehalose repressor TreR and the tre operator.

The trehalose operon of Bacillus subtilis is subject to regulation by induction, mediated by the repressor TreR, and by carbon catabolite repression (CCR). For in vitro investigations, TreR from B. subtilis was overproduced and purified. Its molecular mass, as estimated by SDS-PAGE, is 27 kDa. Size fractionation under native conditions yielded a size estimate of 56 kDa, indicating that TreR exists as a dimer in its native state. Analysis of its interaction with various DNA fragments shows that TreR is able to recognize two tre operators with different efficiencies, and indicates cooperative binding. Previous results have suggested that CCR of the tre operon occurs by a mechanism in which the specific regulator, TreR, may be involved independently of the central component, CcpA. The data presented here indicate that the TreR-tre operator interaction is influenced by several effectors. Thus, the presence of trehalose-6-phosphate, as well as glucose-1-phosphate and sodium chloride, inhibits tre operator binding. Glucose-6-phosphate can act as an anti-inducer, which might reflect its additional role in CCR exerted by glucose.

Analysis of DNA flanking the treA gene of Bacillus subtilis reveals genes encoding a putative specific enzyme IITre and a potential regulator of the trehalose operon.

Nucleotide sequencing revealed the genes treP encoding a putative specific enzyme IITre upstream from treA and treR encoding a potential regulator downstream from treA of Bacillus subtilis 168. The treP gene encodes a 470-amino acid (aa) protein (50 kDa) showing high similarities to several different specific enzymes II of phosphoenolpyruvate-dependent phosphotransferase systems. treR encodes a 238-aa protein (28 kDa) with high homologies in its N-terminal part to DNA-binding proteins including a helixturn-helix motif. Homologies in its C-terminal part place it in the family of FadR-GntR transcriptional regulators. The three genes, treP-treA-treR, are probably organized in one operon expressed by a sigma A-dependent promoter 53 bp upstream from treP and a rho-independent terminator 28 bp downstream from treR.

Expression of the tre operon of Bacillus subtilis 168 is regulated by the repressor TreR.

The tre locus from Bacillus subtilis containing the genes treP, treA, and treR has been analyzed for its regulation. We demonstrate that at least treP and treA form an operon whose expression is regulated at the transcriptional level. TreR activity has been investigated in in vivo and in vitro studies. An insertional inactivation of treR led to a constitutive expression of treP and treA. Upstream of treP we identified a 248-bp DNA fragment containing a potential sigmaA-dependent promoter and two palindromes reflecting potential tre operators which led to complex formation with TreR-containing protein extracts in DNA retardation experiments. This complex formation is abolished in the presence of trehalose-6-phosphate, which probably acts as an inducer. Therefore, we assume that treR encodes the specific Tre repressor involved in regulation of the expression of the tre operon.

Vectors using the phospho-alpha-(1,1)-glucosidase-encoding gene treA of Bacillus subtilis as a reporter.

The intracellular phospho-alpha-(1,1)-glucosidase, TreA, from Bacillus subtilis (Bs) hydrolyses trehalose 6-phosphate into glucose and glucose 6-phosphate. The enzyme is also able to cleave p-nitrophenyl alpha-D-glucopyranoside (PNPG). This enzymatic reaction can be easily monitored in a beta-galactosidase analogous enzyme assay. The vectors we have constructed can be used to study promoter activity in transcriptional treA fusions and may prove especially useful under high-salt conditions due to the halophilic character of TreA. The treA gene is useful as a reporter in either Bs or Escherichia coli (Ec). Such fusions can be integrated in the Bs amyE locus and selected on either kanamycin or chloramphenicol, or used as plasmids in Ec. As an example of the general utility, we demonstrate treA expression under xylA-operator-promoter control.