14-3-3sigma controls mitotic translation to facilitate cytokinesis

14-3-3 proteins are crucial in a wide variety of cellular responses including cell cycle progression, DNA damage checkpoints and apoptosis. One particular 14-3-3 isoform, sigma, is a p53-responsive gene, the function of which is frequently lost in human tumours, including breast and prostate cancers as a result of either hypermethylation of the 14-3-3sigma promoter or induction of an oestrogen-responsive ubiquitin ligase that specifically targets 14-3-3sigma for proteasomal degradation. Loss of 14-3-3sigma protein occurs not only within the tumours themselves but also in the surrounding pre-dysplastic tissue (so-called field cancerization), indicating that 14-3-3sigma might have an important tumour suppressor function that becomes lost early in the process of tumour evolution. The molecular basis for the tumour suppressor function of 14-3-3sigma is unknown. Here we report a previously unknown function for 14-3-3sigma as a regulator of mitotic translation through its direct mitosis-specific binding to a variety of translation/initiation factors, including eukaryotic initiation factor 4B in a stoichiometric manner. Cells lacking 14-3-3sigma, in marked contrast to normal cells, cannot suppress cap-dependent translation and do not stimulate cap-independent translation during and immediately after mitosis. This defective switch in the mechanism of translation results in reduced mitotic-specific expression of the endogenous internal ribosomal entry site (IRES)-dependent form of the cyclin-dependent kinase Cdk11 (p58 PITSLRE), leading to impaired cytokinesis, loss of Polo-like kinase-1 at the midbody, and the accumulation of binucleate cells. The aberrant mitotic phenotype of 14-3-3sigma-depleted cells can be rescued by forced expression of p58 PITSLRE or by extinguishing cap-dependent translation and increasing cap-independent translation during mitosis by using rapamycin. Our findings show how aberrant mitotic translation in the absence of 14-3-3sigma impairs mitotic exit to generate binucleate cells and provides a potential explanation of how 14-3-3sigma-deficient cells may progress on the path to aneuploidy and tumorigenesis.     

Dominant missense mutations in ABCC9 cause Cantú syndrome

Cantú syndrome is characterized by congenital hypertrichosis, distinctive facial features, osteochondrodysplasia and cardiac defects. By using family-based exome sequencing, we identified a de novo mutation in ABCC9. Subsequently, we discovered novel dominant missense mutations in ABCC9 in 14 of the 16 individuals with Cantú syndrome examined. The ABCC9 protein is part of an ATP-dependent potassium (K(ATP)) channel that couples the metabolic state of a cell with its electrical activity. All mutations altered amino acids in or close to the transmembrane domains of ABCC9. Using electrophysiological measurements, we show that mutations in ABCC9 reduce the ATP-mediated potassium channel inhibition, resulting in channel opening. Moreover, similarities between the phenotype of individuals with Cantú syndrome and side effects from the K(ATP) channel agonist minoxidil indicate that the mutations in ABCC9 result in channel opening. Given the availability of ABCC9 antagonists, our findings may have direct implications for the treatment of individuals with Cantú syndrome.     

Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain

Microtubules are cytoskeletal polymers of tubulin involved in many cellular functions. Their dynamic instability is controlled by numerous compounds and proteins, including colchicine and stathmin family proteins. The way in which microtubule instability is regulated at the molecular level has remained elusive, mainly because of the lack of appropriate structural data. Here, we present the structure, at 3.5 A resolution, of tubulin in complex with colchicine and with the stathmin-like domain (SLD) of RB3. It shows the interaction of RB3-SLD with two tubulin heterodimers in a curved complex capped by the SLD amino-terminal domain, which prevents the incorporation of the complexed tubulin into microtubules. A comparison with the structure of tubulin in protofilaments shows changes in the subunits of tubulin as it switches from its straight conformation to a curved one. These changes correlate with the loss of lateral contacts and provide a rationale for the rapid microtubule depolymerization characteristic of dynamic instability. Moreover, the tubulin-colchicine complex sheds light on the mechanism of colchicine's activity: we show that colchicine binds at a location where it prevents curved tubulin from adopting a straight structure, which inhibits assembly.     

Isolation and physical-chemical characterization of a humoral,sleep inducing substance in rabbits (factor ‘delta’)

Zusammenfassung Das Hämodialysat schlafender Kaninchen (unter Einfluss thalamischer Reizung) wurde unter Kontrolle der «Delta-Schlaf»-induzierenden Aktivität fraktioniert. Nach Entsalzung über Sephadex G-10 wurde die hypnogene Fraktion durch anschliessende präparative Dünnschichtchromatographie in 6 verschiedene ninhydrin-positive Fraktionen getrennt (MG>Cl^–). Eine davon enthielt die hypnogene Aktivität. Diese Fraktion 2 wurde über Sephadex G-15 in 3 ninhydrin-positive Gipfel aufgetrennt. Nur die Fraktion mit dem geringstenV _e enthielt die hypnogene Aktivität. Durch Vergleich mit dem Elutionsvolumen von Markersubstanzen konnte für die aktive Substanz ein Molekulargewicht zwischen 355 und 1500 geschätzt werden. Präparative Hochspannungs-Papierelektrophorese trennte das aktive Material in vier Unterfraktionen. Nur eine elektrophoretisch und chromatographisch einheitliche Fraktion (2) enthielt die hypnogene Aktivität. Aus dieser Fraktion wurden nach saurer Hydrolyse mindestens 7 verschiedene Aminosäurereste freigesetzt. Dieses Resultat wurde durch Dansylierung vor und nach saurer Hydrolyse verifiziert; gleichzeitig wurden Artefakte durch kontaminierende Aminosäuren aus dem Papier ausgeschlossen. Die Ergebnisse lassen auf die Peptidnatur einer humoralen «Delta-Schlaf»-induzierenden Substanz schliessen (sleep factor delta).     

Competence to replicate in the unfertilized egg is conferred by Cdc6 during meiotic maturation

Meiotic maturation, the final step of oogenesis, is a crucial stage of development in which an immature oocyte becomes a fertilizable egg. In Xenopus, the ability to replicate DNA is acquired during maturation at breakdown of the nuclear envelope by translation of a DNA synthesis inducer that is not present in the oocyte. Here we identify Cdc6, which is essential for recruiting the minichromosome maintenance (MCM) helicase to the pre-replication complex, as this inducer of DNA synthesis. We show that maternal cdc6 mRNA but not protein is stored in the oocyte. Cdc6 protein is synthesized during maturation, but this process can be blocked by degrading the maternal cdc6 mRNA by oligonucleotide antisense injections or by translation inhibition. Rescue experiments using recombinant Cdc6 protein show that Cdc6 is the only missing replication factor whose translation is necessary and sufficient to confer DNA replication competence to the egg before fertilization. The licence to replicate is given by Cdc6 at the end of meiosis I, but the cytostatic factor (CSF) pathway, which maintains large amounts of active Cdc2/Cyclin B2, prevents the entry into S phase until fertilization.     

Diurnal modulation of pacemaker potentials and calcium current in the mammalian circadian clock

The central biological clock of the mammalian brain is located in the suprachiasmatic nucleus. This hypothalamic region contains neurons that generate a circadian rhythm on a single-cell basis. Clock cells transmit their circadian timing signals to other brain areas by diurnal modulation of their spontaneous firing rate. The intracellular mechanism underlying rhythm generation is thought to consist of one or more self-regulating molecular loops, but it is unknown how these loops interact with the plasma membrane to modulate the ionic conductances that regulate firing behaviour. Here we demonstrate a diurnal modulation of Ca2+ current in suprachiasmatic neurons. This current strongly contributes to the generation of spontaneous oscillations in membrane potential, which occur selectively during daytime and are tightly coupled to spike generation. Thus, day-night modulation of Ca2+ current is a central step in transducing the intracellular cycling of molecular clocks to the rhythm in spontaneous firing rate.     

Utilization of singularity exponent in nearest neighbor based classifier

Classifiers serve as tools for classifying data into classes. They directly or indirectly take a distribution of data points around a given query point into account. To express the distribution of points from the viewpoint of distances from a given point, a probability distribution mapping function is introduced here. The approximation of this function in a form of a suitable power of the distance is presented. How to state this power—the distribution mapping exponent—is described. This exponent is used for probability density estimation in high-dimensional spaces and for classification. A close relation of the exponent to a singularity exponent is discussed. It is also shown that this classifier exhibits better behavior (classification accuracy) than other kinds of classifiers for some tasks.     

Thixotropic effect and electro-osmosis as possible factors of membrane electrical excitability]

Cation motion under the electrical field might break the membrane structure. In the temporary wake of each cation conductivity would be increased. Thus a zone of negative resistance would occur in the intensity-voltage relation. Membrane dehydration, due to electro-osmosis, might induce repeated flip-flops of current at the limits of the negative resistance zone.