RAF inhibitor resistance is mediated by dimerization of aberrantly spliced BRAF(V600E)

Activated RAS promotes dimerization of members of the RAF kinase family. ATP-competitive RAF inhibitors activate ERK signalling by transactivating RAF dimers. In melanomas with mutant BRAF(V600E), levels of RAS activation are low and these drugs bind to BRAF(V600E) monomers and inhibit their activity. This tumour-specific inhibition of ERK signalling results in a broad therapeutic index and RAF inhibitors have remarkable clinical activity in patients with melanomas that harbour mutant BRAF(V600E). However, resistance invariably develops. Here, we identify a new resistance mechanism. We find that a subset of cells resistant to vemurafenib (PLX4032, RG7204) express a 61-kDa variant form of BRAF(V600E), p61BRAF(V600E), which lacks exons 4-8, a region that encompasses the RAS-binding domain. p61BRAF(V600E) shows enhanced dimerization in cells with low levels of RAS activation, as compared to full-length BRAF(V600E). In cells in which p61BRAF(V600E) is expressed endogenously or ectopically, ERK signalling is resistant to the RAF inhibitor. Moreover, a mutation that abolishes the dimerization of p61BRAF(V600E) restores its sensitivity to vemurafenib. Finally, we identified BRAF(V600E) splicing variants lacking the RAS-binding domain in the tumours of six of nineteen patients with acquired resistance to vemurafenib. These data support the model that inhibition of ERK signalling by RAF inhibitors is dependent on levels of RAS-GTP too low to support RAF dimerization and identify a novel mechanism of acquired resistance in patients: expression of splicing isoforms of BRAF(V600E) that dimerize in a RAS-independent manner.     

Making learning critical: Identity, emotion, and power in processes of management development

In recent years, a substantial volume of broadly critical knowledge of management and organization has been established, but comparatively little attention has been given to the question of how the insights of such knowledge might be communicated or applied pedagogically. The temptation or danger, arguably, is for critical knowledge to be substituted for the content of “traditional management education,” with minimal regard for its contribution to processes of personal, social, and organizational development. The chief purpose of this paper is to put some flesh on the claim that critical theory can make an important contribution to the principles and practice of management education. After summarizing some key differences between “traditional management education” and action learning (concerning learning and personal experience), these differences are illustrated by reference to a case study, which in turn, provides a basis for highlighting the relevance of critical thinking for addressing problems and issues thrown up through action learning for managers.     

Allozyme variability, isolation, and dispersal of Eusattus muricatus (Coleoptera: Tenebrionidae) within Silver State Dune Complex, Great Basin, western North America

Genetics of Eusattus muricatus (Coleoptera: Tenebrionidae) populations within Silver State Dune Complex in the northern Great Basin of North America were assayed using allozyme electrophoresis. Hierarchical F -statistics showed no significant population subdivision among 3 valleys comprising Silver State Dune Complex. A regression of estimated pairwise gene flow ( Nm ) against pairwise geographic distance was not significant for populations within these valleys, but it was significant over larger distances. A neighbor-joining phenogram did not represent geographic relationships among populations in the 3 valleys but reflected the past geologic history of these dunes. Pairwise divergence times were low (>7000 yr) for populations within Silver State Dune Complex but were high (>18,000 yr) in outgroups.     

Genetically effective and adult population sizes in the Apache silverspot butterfly, Speyeria nokomis apacheana (Lepidoptera: Mynphalidae)

Genetically effective and adult population sizes of the Apache silverspot butterfly ( Speyeria nokomiis apacheana ) from the Great Basin of North America were tracked from 1991 to 1997. Speyeria nokomis inhabits mesic habitats in the Great Basin, and its recent declines in abundance have raised concerns about its future persistence. Adult population size estimates determined from mark-recapture methods ranged from 100 to 1900 individuals from 1993 through 1997. Genetically effective population sizes based on temporal changes in allozyme frequencies ranged from less tha 1 individual to 20 individuals between 1991 and 1995. Despite these small annual estimates, it is likely that gene flow resulted in overestimation of genetically effective population sizes. These reusuts, combined with observations in the field, suggest that suitable but vacan habitat patches be maintained for potential recolonization by S. n. apacheana.     

Synaptic function modulated by changes in the ratio of synaptotagmin I and IV.

Communication within the nervous system is mediated by Ca2+-triggered fusion of synaptic vesicles with the presynaptic plasma membrane. Genetic and biochemical evidence indicates that synaptotagmin I may function as a Ca2+ sensor in neuronal exocytosis because it can bind Ca2+ and penetrate into lipid bilayers. Chronic depolarization or seizure activity results in the upregulation of a distinct and unusual isoform of the synaptotagmin family, synaptotagmin IV. We have identified a Drosophila homologue of synaptotagmin IV that is enriched on synaptic vesicles and contains an evolutionarily conserved substitution of aspartate to serine that abolishes its ability to bind membranes in response to Ca2+ influx. Synaptotagmin IV forms hetero-oligomers with synaptotagmin I, resulting in synaptotagmin clusters that cannot effectively penetrate lipid bilayers and are less efficient at coupling Ca2+ to secretion in vivo: upregulation of synaptotagmin IV, but not synaptotagmin I, decreases evoked neurotransmission. These findings indicate that modulating the expression of synaptotagmins with different Ca2+-binding affinities can lead to heteromultimers that can regulate the efficiency of excitation-secretion coupling in vivo and represent a new molecular mechanism for synaptic plasticity.     

The DNA sequence and biological annotation of human chromosome 1

The reference sequence for each human chromosome provides the framework for understanding genome function, variation and evolution. Here we report the finished sequence and biological annotation of human chromosome 1. Chromosome 1 is gene-dense, with 3,141 genes and 991 pseudogenes, and many coding sequences overlap. Rearrangements and mutations of chromosome 1 are prevalent in cancer and many other diseases. Patterns of sequence variation reveal signals of recent selection in specific genes that may contribute to human fitness, and also in regions where no function is evident. Fine-scale recombination occurs in hotspots of varying intensity along the sequence, and is enriched near genes. These and other studies of human biology and disease encoded within chromosome 1 are made possible with the highly accurate annotated sequence, as part of the completed set of chromosome sequences that comprise the reference human genome.     

CO2 jets formed by sublimation beneath translucent slab ice in Mars' seasonal south polar ice cap

The martian polar caps are among the most dynamic regions on Mars, growing substantially in winter as a significant fraction of the atmosphere freezes out in the form of CO2 ice. Unusual dark spots, fans and blotches form as the south-polar seasonal CO2 ice cap retreats during spring and summer. Small radial channel networks are often associated with the location of spots once the ice disappears. The spots have been proposed to be simply bare, defrosted ground; the formation of the channels has remained uncertain. Here we report infrared and visible observations that show that the spots and fans remain at CO2 ice temperatures well into summer, and must be granular materials that have been brought up to the surface of the ice, requiring a complex suite of processes to get them there. We propose that the seasonal ice cap forms an impermeable, translucent slab of CO2 ice that sublimates from the base, building up high-pressure gas beneath the slab. This gas levitates the ice, which eventually ruptures, producing high-velocity CO2 vents that erupt sand-sized grains in jets to form the spots and erode the channels. These processes are unlike any observed on Earth.