Mutations of the BRAF gene in human cancer

Cancers arise owing to the accumulation of mutations in critical genes that alter normal programmes of cell proliferation, differentiation and death. As the first stage of a systematic genome-wide screen for these genes, we have prioritized for analysis signalling pathways in which at least one gene is mutated in human cancer. The RAS RAF MEK ERK MAP kinase pathway mediates cellular responses to growth signals. RAS is mutated to an oncogenic form in about 15% of human cancer. The three RAF genes code for cytoplasmic serine/threonine kinases that are regulated by binding RAS. Here we report BRAF somatic missense mutations in 66% of malignant melanomas and at lower frequency in a wide range of human cancers. All mutations are within the kinase domain, with a single substitution (V599E) accounting for 80%. Mutated BRAF proteins have elevated kinase activity and are transforming in NIH3T3 cells. Furthermore, RAS function is not required for the growth of cancer cell lines with the V599E mutation. As BRAF is a serine/threonine kinase that is commonly activated by somatic point mutation in human cancer, it may provide new therapeutic opportunities in malignant melanoma.     

A cryo-electron microscopic study of ribosome-bound termination factor RF2

Protein synthesis takes place on the ribosome, where genetic information carried by messenger RNA is translated into a sequence of amino acids. This process is terminated when a stop codon moves into the ribosomal decoding centre (DC) and is recognized by a class-1 release factor (RF). RFs have a conserved GGQ amino-acid motif, which is crucial for peptide release and is believed to interact directly with the peptidyl-transferase centre (PTC) of the 50S ribosomal subunit. Another conserved motif of RFs (SPF in RF2) has been proposed to interact directly with stop codons in the DC of the 30S subunit. The distance between the DC and PTC is approximately 73 A. However, in the X-ray structure of RF2, SPF and GGQ are only 23 A apart, indicating that they cannot be at DC and PTC simultaneously. Here we show that RF2 is in an open conformation when bound to the ribosome, allowing GGQ to reach the PTC while still allowing SPF-stop-codon interaction. The results indicate new interpretations of accuracy in termination, and have implications for how the presence of a stop codon in the DC is signalled to PTC.     

On the presence of periodic acid schiff positive substances in the paraphysis cerebri,the choroid plexuses and the neuroglia of ambystoma mexicanum

Zusammenfassung Mittels Färbung mit der histochemischen Methode nachMcManus lässt sich eine grosse Menge Glykogen in derParaphysis cerebri vonAmbystoma nachweisen. Glykogentropfen finden sich weiter im Epithel desPlexus chorioideus und vor allem auch in den ependymalen Gliozyten, deren Ausläufer richtige Strassen zum Glykogentransport zu bilden scheinen.Es wird eine Arbeitshypothese aufgestellt, nach der das Glykogen, eine wichtige Energiequelle wie ein Baustein mehr komplexer chemischer Verbindungen, gebildet in der bei Amphibien mächtig entwickelten Paraphyse, in die ventrikuläre Cerebrospinalflüssigkeit gelangt, um von dort aus von den ependymalen Gliozyten aufgenommen und in das nervöse Parenchym hinein transportiert zu werden, wo es eine Rolle bei der Metabolie und dem strukturellen Aufbau spielen kann.

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This investigation was aided by a grant from the Health Organization of the Society for Applied Scientific Research, The Netherlands.     

Ventastega curonica and the origin of tetrapod morphology

The gap in our understanding of the evolutionary transition from fish to tetrapod is beginning to close thanks to the discovery of new intermediate forms such as Tiktaalik roseae. Here we narrow it further by presenting the skull, exceptionally preserved braincase, shoulder girdle and partial pelvis of Ventastega curonica from the Late Devonian of Latvia, a transitional intermediate form between the 'elpistostegids' Panderichthys and Tiktaalik and the Devonian tetrapods (limbed vertebrates) Acanthostega and Ichthyostega. Ventastega is the most primitive Devonian tetrapod represented by extensive remains, and casts light on a part of the phylogeny otherwise only represented by fragmentary taxa: it illuminates the origin of principal tetrapod structures and the extent of morphological diversity among the transitional forms.     

Structure of the Escherichia coli ribosomal termination complex with release factor 2

Termination of protein synthesis occurs when the messenger RNA presents a stop codon in the ribosomal aminoacyl (A) site. Class I release factor proteins (RF1 or RF2) are believed to recognize stop codons via tripeptide motifs, leading to release of the completed polypeptide chain from its covalent attachment to transfer RNA in the ribosomal peptidyl (P) site. Class I RFs possess a conserved GGQ amino-acid motif that is thought to be involved directly in protein-transfer-RNA bond hydrolysis. Crystal structures of bacterial and eukaryotic class I RFs have been determined, but the mechanism of stop codon recognition and peptidyl-tRNA hydrolysis remains unclear. Here we present the structure of the Escherichia coli ribosome in a post-termination complex with RF2, obtained by single-particle cryo-electron microscopy (cryo-EM). Fitting the known 70S and RF2 structures into the electron density map reveals that RF2 adopts a different conformation on the ribosome when compared with the crystal structure of the isolated protein. The amino-terminal helical domain of RF2 contacts the factor-binding site of the ribosome, the 'SPF' loop of the protein is situated close to the mRNA, and the GGQ-containing domain of RF2 interacts with the peptidyl-transferase centre (PTC). By connecting the ribosomal decoding centre with the PTC, RF2 functionally mimics a tRNA molecule in the A site. Translational termination in eukaryotes is likely to be based on a similar mechanism.     

Reversible necrosis at the end-plate region in striated muscles of the rat poisoned with cholinesterase inhibitors

Résumé Après l'injection d'un toxique anticholinesté-rasique à dose presque léthale, une nécrose focale des fibres musculaires striées du rat se développe en 24 h. 10 jours après l'injection, la régénération est presque totale. Les parties nécrotiques se trouvent dans les régions des plaques motrices. Les résultats obtenus suggèrent que l'action dépolarisante de l'acétylcholine au niveau des plaques motrices des muscles actifs, augmentée et prolongée par l'intoxication, est responsable de la nécrose.     

Synapses of the ribbon type in the pineal organ ofLacerta vivipara (reptiles,lacertilians)

Résumé Un appareil photorécepteur et neuroconducteur est hautement différencié mais quantitativement peu développé dans l'organe pinéal deLacerta vivipara adulte. L'existence de rubans synaptiques typiques, à la jonction des photorécepteurs et des cellules ganglionnaires est démontrée.

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Acknowledgment: our thanks are due to MrsS. Vernerey for her excellent technical assistance.     

Mutations in SLC20A2 link familial idiopathic basal ganglia calcification with phosphate homeostasis

Familial idiopathic basal ganglia calcification (IBGC) is a genetic condition with a wide spectrum of neuropsychiatric symptoms, including parkinsonism and dementia. Here, we identified mutations in SLC20A2, encoding the type III sodium-dependent phosphate transporter 2 (PiT2), in IBGC-affected families of varied ancestry, and we observed significantly impaired phosphate transport activity for all assayed PiT2 mutants in Xenopus laevis oocytes. Our results implicate altered phosphate homeostasis in the etiology of IBGC.     

From parasympathomimetics to parasympatholytics by homologous substitution

Zusammenfassung Auf Grund der theoretischen Befunde war zu erwarten, dass Änderungen an der negativen und positiven Seite der Dioxolan-Verbindung 2249 F zu graduellen Übergängen von Parasympathikomimetika zu Parasympathikolytika führen würden. Die Experimente bestätigten die Erwartungen vollauf.Untersuchung homologer Reihen von Alkyltrialkylammonium-Verbindungen zeigte die negative Gruppe als nicht wesentlich für die parasympathische Wirkung und ergab ebenso einen graduellen Übergang von Mimetika zu Lytika.