THM1 negatively modulates mouse sonic hedgehog signal transduction and affects retrograde intraflagellar transport in cilia

Characterization of previously described intraflagellar transport (IFT) mouse mutants has led to the proposition that normal primary cilia are required for mammalian cells to respond to the sonic hedgehog (SHH) signal. Here we describe an N-ethyl-N-nitrosourea-induced mutant mouse, alien (aln), which has abnormal primary cilia and shows overactivation of the SHH pathway. The aln locus encodes a novel protein, THM1 (tetratricopeptide repeat-containing hedgehog modulator-1), which localizes to cilia. aln-mutant cilia have bulb-like structures at their tips in which IFT proteins (such as IFT88) are sequestered, characteristic of Chlamydomonas reinhardtii and Caenorhabditis elegans retrograde IFT mutants. RNA-interference knockdown of Ttc21b (which we call Thm1 and which encodes THM1) in mouse inner medullary collecting duct cells expressing an IFT88-enhanced yellow fluorescent protein fusion recapitulated the aln-mutant cilial phenotype, and live imaging of these cells revealed impaired retrograde IFT. In contrast to previously described IFT mutants, Smoothened and full-length glioblastoma (GLI) proteins localize to aln-mutant cilia. We hypothesize that the aln retrograde IFT defect causes sequestration of IFT proteins in aln-mutant cilia and leads to the overactivated SHH signaling phenotype. Specifically, the aln mutation uncouples the roles of anterograde and retrograde transport in SHH signaling, suggesting that anterograde IFT is required for GLI activation and that retrograde IFT modulates this event.     

Stratigraphy and paleontology of the marine Jurrassic strata in Vietnam

Marine Jurassic strata in Vietnam have yielded characteristic ammonites and occur in: (1) west Quang Binh Province located in the eastern margin of the Nam Theun Basin (the main part of which is located in Middle Laos); (2) the Nong Son Basin situated in the north of the Kon Tum Geoblock; (3) the Da Lat Basin, the largest Jurassic basin in Vietnam, extending from the southern margin of the Kon Tum Geoblock to southeast of Ho Chi Minh City. In the first two, Lower Jurassic marine beds occupy the lower part of the section and grade up into Middle Jurassic red beds. In central Da Lat Basin the section consists completely of marine strata containing many ammonite levels dated from Early Sinemurian to Bajocian, whereas the marginal basin section is like the first two areas. Based particularly on biostratigraphical evidence, a marine transgression is considered to have entered the Indochina Block as early as the Late Hettangian. During the early transgression the fauna was distinctly endemic, showing the separate character of the basins, but since the Late Sinemurian, communication between the Da Lat and other basins was established; the marine regime in Da Lat Basin ended around the Late Bathonian.     

FKF1 is essential for photoperiodic-specific light signalling in Arabidopsis

Adaptation to seasonal change is a crucial component of an organism's survival strategy. To monitor seasonal variation, organisms have developed the capacity to measure day length (photoperiodism). Day-length assessment involves the photoperiodic control of flowering in Arabidopsis thaliana, whereby the coincidence of light and high expression of CONSTANS (CO) induces the expression of FLOWERING LOCUS T (FT), leading to flowering in long-day conditions. Although controlling CO expression is clearly a key step in day-length discrimination, the mechanism that generates day-length-dependent CO expression remains unknown. Here we show that the clock-controlled FLAVIN-BINDING, KELCH REPEAT, F-BOX (FKF1) protein has an essential role in generating the diurnal CO peak and that this function is dependent on light. We show that a recombinant FKF1 LIGHT, OXYGEN OR VOLTAGE (LOV) domain binds the chromophore flavin mononucleotide and undergoes light-induced photochemistry, indicating that FKF1 may function as a photoperiodic blue-light receptor. It is likely that the circadian control of FKF1 expression and the light regulation of FKF1 function coincide to control the daytime CO waveform precisely, which in turn is crucial for day-length discrimination by Arabidopsis.     

Bone recognition mechanism of porcine osteocalcin from crystal structure

Osteocalcin is the most abundant noncollagenous protein in bone, and its concentration in serum is closely linked to bone metabolism and serves as a biological marker for the clinical assessment of bone disease. Although its precise mechanism of action is unclear, osteocalcin influences bone mineralization, in part through its ability to bind with high affinity to the mineral component of bone, hydroxyapatite. In addition to binding to hydroxyapatite, osteocalcin functions in cell signalling and the recruitment of osteoclasts and osteoblasts, which have active roles in bone resorption and deposition, respectively. Here we present the X-ray crystal structure of porcine osteocalcin at 2.0 A resolution, which reveals a negatively charged protein surface that coordinates five calcium ions in a spatial orientation that is complementary to calcium ions in a hydroxyapatite crystal lattice. On the basis of our findings, we propose a model of osteocalcin binding to hydroxyapatite and draw parallels with other proteins that engage crystal lattices.     

Hydrodynamics and heat transfer of gas-solid two-phase mixtures flowing through packed beds–a review

Flow of a gas-solid two-phase mixture through a packed bed is relevant to a number of industrial processes such as heat recovery and filtration of dusty flue gases, iron making in shaft reactors, gas purification, and sorption enhanced reaction processes. In spite of the industrial relevance, little work has been reported in the literature. The limited amount of research work has mainly addressed the mac- roscopic hydrodynamics in terms of pressure drop and solids hold-ups at the ambient temperature. Very little is done, until fairly recent, on solids motion at the single particle level, hydrodynamics at elevated temperatures and heat transfer. This paper reviews the recent development in the field including both the hydrodynamics and heat transfer of gas-solid two-phase mixtures flowing through packed beds, which is believed to represent the state-of-the-art in the field. The review is not aimed to be exhaustive but rather focused on our own work carried out over the past few years in the Institute of Particle Science ＆ Engineering at the University of Leeds. And some of our results are compared with that of other groups.     

X-ray structures of general anaesthetics bound to a pentameric ligand-gated ion channel

General anaesthetics have enjoyed long and widespread use but their molecular mechanism of action remains poorly understood. There is good evidence that their principal targets are pentameric ligand-gated ion channels (pLGICs) such as inhibitory GABA(A) (γ-aminobutyric acid) receptors and excitatory nicotinic acetylcholine receptors, which are respectively potentiated and inhibited by general anaesthetics. The bacterial homologue from Gloeobacter violaceus (GLIC), whose X-ray structure was recently solved, is also sensitive to clinical concentrations of general anaesthetics. Here we describe the crystal structures of the complexes propofol/GLIC and desflurane/GLIC. These reveal a common general-anaesthetic binding site, which pre-exists in the apo-structure in the upper part of the transmembrane domain of each protomer. Both molecules establish van der Waals interactions with the protein; propofol binds at the entrance of the cavity whereas the smaller, more flexible, desflurane binds deeper inside. Mutations of some amino acids lining the binding site profoundly alter the ionic response of GLIC to protons, and affect its general-anaesthetic pharmacology. Molecular dynamics simulations, performed on the wild type (WT) and two GLIC mutants, highlight differences in mobility of propofol in its binding site and help to explain these effects. These data provide a novel structural framework for the design of general anaesthetics and of allosteric modulators of brain pLGICs.