Specificity in Toll-like receptor signalling through distinct effector functions of TRAF3 and TRAF6

Toll-like receptors (TLRs) are activated by pathogen-associated molecular patterns to induce innate immune responses and production of pro-inflammatory cytokines, interferons and anti-inflammatory cytokines. TLRs activate downstream effectors through adaptors that contain Toll/interleukin-1 receptor (TIR) domains, but the mechanisms accounting for diversification of TLR effector functions are unclear. To dissect biochemically TLR signalling, we established a system for isolating signalling complexes assembled by dimerized adaptors. Using MyD88 as a prototypical adaptor, we identified TNF receptor-associated factor 3 (TRAF3) as a new component of TIR signalling complexes that is recruited along with TRAF6. Using myeloid cells from TRAF3- and TRAF6-deficient mice, we show that TRAF3 is essential for the induction of type I interferons (IFN) and the anti-inflammatory cytokine interleukin-10 (IL-10), but is dispensable for expression of pro-inflammatory cytokines. In fact, TRAF3-deficient cells overproduce pro-inflammatory cytokines owing to defective IL-10 production. Despite their structural similarity, the functions of TRAF3 and TRAF6 are largely distinct. TRAF3 is also recruited to the adaptor TRIF (Toll/IL-1 receptor domain-containing adaptor-inducing IFN-beta) and is required for marshalling the protein kinase TBK1 (also called NAK) into TIR signalling complexes, thereby explaining its unique role in activation of the IFN response.     

Active-site remodelling in the bifunctional fructose-1,6-bisphosphate aldolase/phosphatase

Fructose-1,6-bisphosphate (FBP) aldolase/phosphatase is a bifunctional, thermostable enzyme that catalyses two subsequent steps in gluconeogenesis in most archaea and in deeply branching bacterial lineages. It mediates the aldol condensation of heat-labile dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (GAP) to FBP, as well as the subsequent, irreversible hydrolysis of the product to yield the stable fructose-6-phosphate (F6P) and inorganic phosphate; no reaction intermediates are released. Here we present a series of structural snapshots of the reaction that reveal a substantial remodelling of the active site through the movement of loop regions that create different catalytic functionalities at the same location. We have solved the three-dimensional structures of FBP aldolase/phosphatase from thermophilic Thermoproteus neutrophilus in a ligand-free state as well as in complex with the substrates DHAP and FBP and the product F6P to resolutions up to 1.3??. In conjunction with mutagenesis data, this pinpoints the residues required for the two reaction steps and shows that the sequential binding of additional Mg(2+) cations reversibly facilitates the reaction. FBP aldolase/phosphatase is an ancestral gluconeogenic enzyme optimized for high ambient temperatures, and our work resolves how consecutive structural rearrangements reorganize the catalytic centre of the protein to carry out two canonical reactions in a very non-canonical type of bifunctionality.     

A linear forecasting model and its application to economic data

We present a forecasting model based on fuzzy pattern recognition and weighted linear regression. In this model fuzzy pattern recognition is used to find homogeneous fuzzy classes in a heterogeneous data set. It is assumed that the classes represent typical situations. For each class a weighted regression analysis is conducted. The forecasting results obtained by the class regression analysis are aggregated to obtain the ‘overall’ estimation of the regression model. We apply the model to the forecasting of economic data of the USA. Copyright © 2001 John Wiley & Sons, Ltd.     

A Cambrian micro-lobopodian and the evolution of arthropod locomotion and reproduction

The evolutionary success of arthropods, the most abundant and diverse animal group, is mainly based on their segmented body and jointed appendages, features that had evolved most likely already before the Cambrian. The first arthropod-like animals, the lobopodians from the Early Cambrian, were unsclerotized and worm-like, and they had unjointed tubular legs. Here we describe the first three-dimensionally preserved Cambrian lobopodian. The material presented of Orstenotubulus evamuellerae gen. et sp. nov. is the smallest and youngest of a lobopodian known. O. evamuellerae shows strikingly detailed similarities to Recent tardigrades and/or onychophorans in its cellular-structured cuticle and the telescopic spines. It also shows similarities to other, longer known lobopodians, but which are ten times as large as the new form. These similarities include the finely annulated body and legs, which is characteristic also for Recent onychophorans, and paired humps continuing into spines situated dorsally to the leg insertions, a feature lacking in the extant forms. The morphology of O. evamuellerae not only elucidates our knowledge about lobopodians, but also aids in a clearer picture of the early evolution of arthropods. An example is the single ventral gonopore between a limb pair of O. evamuellerae, which indicates that a single gonopore, as developed in onychophorans, tardigrades, pentastomids, myriapods and insects, might represent the plesiomorphic state for Arthropoda, while the paired state in chelicerates and crustaceans was convergently achieved. Concerning life habits, the lateral orientation of the limbs and their anchoring spines of the new lobopodian imply that early arthropods were crawlers rather than walkers.     

An oxygenase that forms and deoxygenates toxic epoxide

Catabolism may give rise to toxic intermediates that compromise cell vitality, such as epoxide formation in the recently elucidated and apparently universal bacterial coenzyme A (CoA)-dependent degradation of phenylacetic acid. This compound is central to the catabolism of a variety of aromatics, such as phenylalanine, lignin-related compounds or environmental contaminants. The key phenylacetyl-CoA monooxygenase (epoxidase) of the pathway, PaaABCE, is also connected to the production of various primary and secondary metabolites, as well as to the virulence of certain pathogens. However, the enzyme complex has so far not been investigated in detail. Here we characterize the bacterial multicomponent monooxygenase PaaABCE that, surprisingly, not only transforms phenylacetyl-CoA into its ring-1,2-epoxide, but also mediates the NADPH-dependent removal of the epoxide oxygen, regenerating phenylacetyl-CoA with formation of water. We provide evidence for a catalytic di-iron centre that is probably the key to the unprecedented deoxygenation of an organic compound by an oxygenase. Presumably, the bifunctionality is vital to avoid toxic intracellular epoxide levels if the subsequent catabolic steps are impeded. Our data suggest that detoxification is assisted by two thioesterases (PaaI and PaaY) forming non-reactive breakdown products. Hence, PaaABCE may harbour an intrinsic escape mechanism from its own toxic product and represents the archetype of a bifunctional oxygenase/deoxygenase. Analogous reactions may possibly be catalysed by other di-iron epoxidases.     

Histamine: Entering physiology

Zusammenfassung Die hier geschilderte Periode der Histaminforschung ist durch die Erkenntnis gekennzeichnet, dass grosse Veränderungen in der Geschwindigkeit der Histaminbildung unter physiologischen Verhältnissen vorkommen. In der Magenmucosa bedingen Gastrin und Nahrungszufuhr eine Mobilisierung von Histamin und eine Erhöhung der Aktivität der Histidindecarboxylase. Bei verschiedenen Formen von normalem und malignem Wachstum wird in den Geweben «Nascent-Histamin» gebildet, dessen Wirkung exogenes Histamin nicht ausüben kann und auch nicht mit Antihistaminen antagonisiert wird⁶⁵.     

Deficiency of UBR1, a ubiquitin ligase of the N-end rule pathway, causes pancreatic dysfunction, malformations and mental retardation (Johanson-Blizzard syndrome)

Johanson-Blizzard syndrome (OMIM 243800) is an autosomal recessive disorder that includes congenital exocrine pancreatic insufficiency, multiple malformations such as nasal wing aplasia, and frequent mental retardation. We mapped the disease-associated locus to chromosome 15q14-21.1 and identified mutations, mostly truncating ones, in the gene UBR1 in 12 unrelated families with Johanson-Blizzard syndrome. UBR1 encodes one of at least four functionally overlapping E3 ubiquitin ligases of the N-end rule pathway, a conserved proteolytic system whose substrates include proteins with destabilizing N-terminal residues. Pancreas of individuals with Johanson-Blizzard syndrome did not express UBR1 and had intrauterine-onset destructive pancreatitis. In addition, we found that Ubr1(-/-) mice, whose previously reported phenotypes include reduced weight and behavioral abnormalities, had an exocrine pancreatic insufficiency, with impaired stimulus-secretion coupling and increased susceptibility to pancreatic injury. Our findings indicate that deficiency of UBR1 perturbs the pancreas' acinar cells and other organs, presumably owing to metabolic stabilization of specific substrates of the N-end rule pathway.     

Gene-culture coevolution between cattle milk protein genes and human lactase genes

Milk from domestic cows has been a valuable food source for over 8,000 years, especially in lactose-tolerant human societies that exploit dairy breeds. We studied geographic patterns of variation in genes encoding the six most important milk proteins in 70 native European cattle breeds. We found substantial geographic coincidence between high diversity in cattle milk genes, locations of the European Neolithic cattle farming sites (>5,000 years ago) and present-day lactose tolerance in Europeans. This suggests a gene-culture coevolution between cattle and humans.     

Eccentricities of hydrocarbons on my way from the Diyls to the Ylids

Zusammenfassung In dem vorliegenden Artikel werden Kohlenwasserstoffe behandelt, die — im Laufe einer etwa 45 jährigen Forschung des Autors synthetisiert — entweder in ihrer Bildungsweise oder in ihrer Struktur oder schliesslich in ihren Eigenschaften ein wenig skurril sind.In Modellverbindungen wurde der Einfluss der Spannung auf die Tendenz der Ringöffnung zu Diradikalen (kurz Diyle genannt) untersucht und weiterhin am Tschitschibabinschen Kohlenwasserstoff und Analogen die Alternative studiert, ob es sich um eine Valenztautomerie oder Mesomerie zwischen chinoiden Derivaten und Diradikalen handelt.Während die anstehenden Probleme hier noch nicht restlos geklärt sind, zeigen die , -Tetraphenylpolyene auch mit längerer Polyenkette, ferner das -Carotin keinen Diylcharakter. Auch Tetraphenyl-dihydrophenanthren9 und das aromatische Propellan12 sind inert gegenüber Sauerstoff.In diesem Zusammenhang wurde über die Präparierung von Cyclopolyenen wie19 und22 berichtet. Besonderes Interesse bietet das auf der Ammoniumylid-Basis bereitete Tetraphenylcyclododecahexaen27, das beim Erhitzen über das Cyclobutanderivat28 zum Phenanthren zerfällt. Von den ebenfalls zu den Cyclopolyenen gehörenden beiden stereoisomereno-Hexaphenylenen30A und30B konnte den Erwartungen gemäss30B in optische Antipoden zerlegt werden. Tetrabenzonaphthalin31, in dessen Molekül die mittlere C=C-Bindung besonders reaktiv ist, bildete mit Phenanthrenchinon beim Bestrahlen das Propellan32, das beim Erhitzen wieder in die Ausgangskomponenten zerfiel.Abschliessend wird über die Bildungsweise von Dehydrobenzol und seine Struktur berichtet und das dienophile Verhalten des nur kurzlebien Kohlenwasserstoffs aufgezeigt. Zum Unterschied von Dehydrobenzol enthalten die niedergliedrigen Cycloalkine eine echte Acetylenbindung. Sie sind ebenfalls kurzlebig; das cyclohexin tetramerisiert sich über sein nicht-isolierbares Cyclobutadien-Derivat.