Validation tools: can they indicate the information content of macromolecular crystal structures?

AbstractThe explosive increase in the number of published three-dimensionsal structures of macromolecules determined by X-ray analysis places a responsibility on experimentalists, referees and curators of databases to ensure correspondence between the structure parameters and data. Validation tools will evolve as more appropriate statistical techniques and new information, such as that from proteins analysed at atomic resolution, becomes available

Ethnobotany of the Samburu of Mt. Nyiru, South Turkana, Kenya

Traditional plant use is of extremely high importance in many societies, and prevalent in African communities. This knowledge is however dwindling rapidly due to changes towards a more Western lifestyle. The influence of modern tourism cannot be neglected in this context. This paper examines the plant use of the Samburu of the Mt. Nyiru area in Northern Kenya. The Samburu pastoralists of Kenya are still amongst the most traditional communities of the country and have retained most of their knowledge about the use of a large part of the plants in their environment for a wide variety of purposes. The results indicate that the local population has a very high knowledge of the plants in their surroundings, and attributes a purpose to a large percentage of the plants found. 448 plant species were collected, identified and their Samburu names and traditional uses recorded. 199 species were reported as of "no use". The high proportion of 249 plant species however had some traditional use: The highest number (180 species) was used as fodder, followed by 80 species that had medicinal use. Firewood (59 species), construction (42 species), tools (31 species), food (29 species) and ceremonial use (19 species) ranked far behind. Traditionally the Samburu attribute most illnesses to the effect of pollutants that block or inhibit digestion. This can include "polluted" food, contagion through sick people as well as witchcraft. In most cases the treatment of illness involves herbal purgatives to cleanse the patient. There are however frequent indications of plant use for common problems like wounds, parasites, body aches and burns. The change from a nomadic to a more sedentary lifestyle, often observed in other areas of the country, has affected the Samburu of remote Mt. Nyiru to a much lesser extent and did so far not lead to a major loss of traditional plant knowledge. However, overgrazing and over-exploitation of plant resources have already led to a decline of the plant material available

The structural basis for selective binding of non-methylated CpG islands by the CFP1 CXXC domain

CFP1 is a CXXC domain-containing protein and an essential component of the SETD1 histone H3K4 methyltransferase complex. CXXC domain proteins direct different chromatin-modifying activities to various chromatin regions. Here, we report crystal structures of the CFP1 CXXC domain in complex with six different CpG DNA sequences. The crescent-shaped CFP1 CXXC domain is wedged into the major groove of the CpG DNA, distorting the B-form DNA, and interacts extensively with the major groove of the DNA. The structures elucidate the molecular mechanism of the non-methylated CpG-binding specificity of the CFP1 CXXC domain. The CpG motif is confined by a tripeptide located in a rigid loop, which only allows the accommodation of the non-methylated CpG dinucleotide. Furthermore, we demonstrate that CFP1 has a preference for a guanosine nucleotide following the CpG motif

A CI-Independent Form of Replicative Inhibition: Turn Off of Early Replication of Bacteriophage Lambda

Several earlier studies have described an unusual exclusion phenotype exhibited by cells with plasmids carrying a portion of the replication region of phage lambda. Cells exhibiting this inhibition phenotype (IP) prevent the plating of homo-immune and hybrid hetero-immune lambdoid phages. We have attempted to define aspects of IP, and show that it is directed to repλ phages. IP was observed in cells with plasmids containing a λ DNA fragment including oop, encoding a short OOP micro RNA, and part of the lambda origin of replication, oriλ, defined by iteron sequences ITN1-4 and an adjacent high AT-rich sequence. Transcription of the intact oop sequence from its promoter, pO is required for IP, as are iterons ITN3–4, but not the high AT-rich portion of oriλ. The results suggest that IP silencing is directed to theta mode replication initiation from an infecting repλ genome, or an induced repλ prophage. Phage mutations suppressing IP, i.e., Sip, map within, or adjacent to cro or in O, or both. Our results for plasmid based IP suggest the hypothesis that there is a natural mechanism for silencing early theta-mode replication initiation, i.e. the buildup of λ genomes with oop+ oriλ+ sequence

Self Assessment in Insects: Honeybee Queens Know Their Own Strength

Contests mediate access to reproductive opportunities in almost all species of animals. An important aspect of the evolution of contests is the reduction of the costs incurred during intra-specific encounters to a minimum. However, escalated fights are commonly lethal in some species like the honeybee, Apis mellifera. By experimentally reducing honeybee queens' fighting abilities, we demonstrate that they refrain from engaging in lethal contests that typically characterize their reproductive dominance behavior and coexist peacefully within a colony. This suggests that weak queens exploit an alternative reproductive strategy and provides an explanation for rare occurrences of queen cohabitation in nature. Our results further indicate that self-assessment, but not mutual assessment of fighting ability occurs prior to and during the agonistic encounters

Unexpected large evasion fluxes of carbon dioxide from turbulent streams draining the world’s mountains

Inland waters, including streams and rivers, are active components of the global carbon cycle. Despite the large areal extent of the world’s mountains, the role of mountain streams for global carbon fluxes remains elusive. Using recent insights from gas exchange in turbulent streams, we found that areal CO2 evasion fluxes from mountain streams equal or exceed those reported from tropical and boreal streams, typically regarded as hotspots of aquatic carbon fluxes. At the regional scale of the Swiss Alps, we present evidence that emitted CO2 derives from lithogenic and biogenic sources within the catchment and delivered by the groundwater to the streams. At a global scale, we estimate the CO2 evasion from mountain streams to 167 ± 1.5 Tg C yr−1, which is high given their relatively low areal contribution to the global stream and river networks. Our findings shed new light on mountain streams for global carbon fluxes

Intervening with Urinary Tract Infections Using Anti-Adhesives Based on the Crystal Structure of the FimH–Oligomannose-3 Complex

Escherichia coli strains adhere to the normally sterile human uroepithelium using type 1 pili, that are long, hairy surface organelles exposing a mannose-binding FimH adhesin at the tip. A small percentage of adhered bacteria can successfully invade bladder cells, presumably via pathways mediated by the high-mannosylated uroplakin-Ia and alpha3beta1 integrins found throughout the uroepithelium. Invaded bacteria replicate and mature into dense, biofilm-like inclusions in preparation of fluxing and of infection of neighbouring cells, being the major cause of the troublesome recurrent urinary tract infections.We demonstrate that alpha-D-mannose based inhibitors of FimH not only block bacterial adhesion on uroepithelial cells but also antagonize invasion and biofilm formation. Heptyl alpha-D-mannose prevents binding of type 1-piliated E. coli to the human bladder cell line 5637 and reduces both adhesion and invasion of the UTI89 cystitis isolate instilled in mouse bladder via catheterization. Heptyl alpha-D-mannose also specifically inhibited biofilm formation at micromolar concentrations. The structural basis of the great inhibitory potential of alkyl and aryl alpha-D-mannosides was elucidated in the crystal structure of the FimH receptor-binding domain in complex with oligomannose-3. FimH interacts with Man alpha1,3Man beta1,4GlcNAc beta1,4GlcNAc in an extended binding site. The interactions along the alpha1,3 glycosidic bond and the first beta1,4 linkage to the chitobiose unit are conserved with those of FimH with butyl alpha-D-mannose. The strong stacking of the central mannose with the aromatic ring of Tyr48 is congruent with the high affinity found for synthetic inhibitors in which this mannose is substituted for by an aromatic group.The potential of ligand-based design of antagonists of urinary tract infections is ruled by the structural mimicry of natural epitopes and extends into blocking of bacterial invasion, intracellular growth and capacity to fluxing and of recurrence of the infection

The CCP4 suite: integrative software for macromolecular crystallography

The Collaborative Computational Project No. 4 (CCP4) is a UK-led international collective with a mission to develop, test, distribute and promote software for macromolecular crystallography. The CCP4 suite is a multiplatform collection of programs brought together by familiar execution routines, a set of common libraries and graphical interfaces. The CCP4 suite has experienced several considerable changes since its last reference article, involving new infrastructure, original programs and graphical interfaces. This article, which is intended as a general literature citation for the use of the CCP4 software suite in structure determination, will guide the reader through such transformations, offering a general overview of the new features and outlining future developments. As such, it aims to highlight the individual programs that comprise the suite and to provide the latest references to them for perusal by crystallographers around the world

Bundles of acceleration on Banach manifolds

We consider an infinite dimensional manifold M modelled on a Banach space E and we construct smooth fiber bundle structures on the tangent bundle of order two T^2M, which consists of all smooth curves of M that agree up to their acceleration, as well as on the corresponding second order frame bundle L^2M. These bundles prove to be associated with respect to the identity representation of the general linear group GL(E}) that serves as the structure group of both of them. Moreover, a bijective correspondence between linear connections on T^2M and connection forms of L^2M is revealed

A generalized second order frame bundle for Fréchet manifolds

Working within the framework of Fréchet modelled infinite dimensional manifolds, we propose a generalized notion of second order frame bundle. We revise in this way the classical notion of bundles of linear frames of order two, the direct definition and study of which is problematic due to intrinsic difficulties of the space models. However, this new structure keeps all the fundamental characteristics of a frame bundle: It is a principal Frechet bundle associated (differentially and geometrically) with the corresponding second order tangent bundle