Peptide binding to class I MHC on living cells and quantitation of complexes required for CTL lysis

Antigenic peptides are presented to CD8+T lymphocytes by class I major histocompatibility complex (MHC) molecules. Peptides specifically bind to purified class I molecules in vitro, and to class I molecules on cells at nonphysiological temperatures. We report here the kinetic and equilibrium parameters for the binding of radiolabelled influenza nucleoprotein peptides (NP-Y365-380 and shorter homologues) to the murine H-2Db molecule on intact, viable cells at 37 degrees C. In contrast to earlier reports, we show that peptide binding is rapid and reversible, with dissociation constants ranging from nanomolar to micromolar, suggestive of typical ligand-receptor interactions. Only 10% of cell-surface Db molecules can bind these peptides. To address the relationship between peptide binding and T-cell recognition of the antigen-MHC complex, we determined the minimum number of complexes required to sensitize a target cell for lysis by class I-restricted cytotoxic T-lymphocytes. Our data indicate that EL4 thymoma cells (H-2b) can be sensitized for lysis by cytotoxic T-lymphocytes when as few as 200 class I-peptide complexes (less than 0.08% of surface Db molecules) are present per cell.     

Point mutation of an FGF receptor abolishes phosphatidylinositol turnover and Ca2+ flux but not mitogenesis.

Stimulation of certain receptor tyrosine kinases results in the tyrosine phosphorylation and activation of phospholipase C gamma (PLC gamma), an enzyme that catalyses the hydrolysis of phosphatidylinositol (PtdIns). This hydrolysis generates diacylglycerol and free inositol phosphate, which in turn activate protein kinase C and increase intracellular Ca2+, respectively. PLC gamma physically associates with activated receptor tyrosine kinases, suggesting that it is a substrate for direct phosphorylation by these kinases. Here we report that a fibroblast growth factor (FGF) receptor with a single point mutation at residue 766 replacing tyrosine with phenylalanine fails to associate with PLC gamma in response to FGF. This mutant receptor also failed to mediate PtdIns hydrolysis and Ca2+ mobilization after FGF stimulation. However, the mutant receptor phosphorylated itself and several other cellular proteins, and it mediated mitogenesis in response to FGF. These findings show that a point mutation in the FGF receptor selectively eliminates activation of PLC gamma and that neither Ca2+ mobilization nor PtdIns hydrolysis are required for FGF-induced mitogenesis.     

Nova regulates brain-specific splicing to shape the synapse

Alternative RNA splicing greatly increases proteome diversity and may thereby contribute to tissue-specific functions. We carried out genome-wide quantitative analysis of alternative splicing using a custom Affymetrix microarray to assess the role of the neuronal splicing factor Nova in the brain. We used a stringent algorithm to identify 591 exons that were differentially spliced in the brain relative to immune tissues, and 6.6% of these showed major splicing defects in the neocortex of Nova2-/- mice. We tested 49 exons with the largest predicted Nova-dependent splicing changes and validated all 49 by RT-PCR. We analyzed the encoded proteins and found that all those with defined brain functions acted in the synapse (34 of 40, including neurotransmitter receptors, cation channels, adhesion and scaffold proteins) or in axon guidance (8 of 40). Moreover, of the 35 proteins with known interaction partners, 74% (26) interact with each other. Validating a large set of Nova RNA targets has led us to identify a multi-tiered network in which Nova regulates the exon content of RNAs encoding proteins that interact in the synapse.     

Muted climate variations in continental Siberia during the mid-Pleistocene epoch

The large difference in carbon and oxygen isotope data from the marine record between marine oxygen isotope stage 12 (MIS 12) and MIS 11, spanning the interval between about 480 and 380 kyr ago, has been interpreted as a transition between an extremely cold glacial period and an unusually warm interglacial period, with consequences for global ice volume, sea level and the global carbon cycle. The extent of the change is intriguing, because orbital forcing is predicted to have been relatively weak at that time. Here we analyse a continuous sediment record from Lake Baikal, Siberia, which reveals a virtually continuous interglacial diatom assemblage, a stable littoral benthic diatom assemblage and lithogenic sediments with 'interglacial' characteristics for the period from MIS 15a to MIS 11 (from about 580 to 380 kyr ago). From these data, we infer significantly weaker climate contrasts between MIS 12 and 11 than during more recent glacial-interglacial transitions in the late Pleistocene epoch (about 130 to 10 kyr ago). For the period from MIS 15a to MIS 11, we also infer an apparent lack of extensive mountain glaciation.     

Ecology: Darwin's naturalization hypothesis challenged

Naturalized plants can have a significant ecological and economic impact, yet they comprise only a fraction ot the plant species introduced by humans. Darwin proposed that introduced plant species will be less likely to establish a self-sustaining wild population in places with congeneric native species because the introduced plants have to compete with their close relatives, or are more likely to be attacked by native herbivores or pathogens, a theory known as Darwin's naturalization hypothesis. Here we analyze a complete list of seed-plant species that have been introduced to New Zealand and find that those with congeneric relatives are significantly more, not less, likely to naturalize--perhaps because they share with their native relatives traits that pre-adapt them to their new environment.     

Why stainless steel corrodes

Stainless steels are used in countless diverse applications for their corrosion resistance. Although they have extremely good general resistance, they are nevertheless susceptible to pitting corrosion. This localized dissolution of an oxide-covered metal in specific aggressive environments is one of the most common and catastrophic causes of failure of metallic structures. The pitting process has been described as random, sporadic and stochastic and the prediction of the time and location of events remains extremely difficult. Many contested models of pitting corrosion exist, but one undisputed aspect is that manganese sulphide inclusions play a critical role. Indeed, the vast majority of pitting events are found to occur at, or adjacent to, such second-phase particles. Chemical changes in and around sulphide inclusions have been postulated as a mechanism for pit initiation but such variations have never been measured. Here we use nanometre-scale secondary ion mass spectroscopy to demonstrate a significant reduction in the Cr:Fe ratio of the steel matrix around MnS particles. These chromium-depleted zones are susceptible to high-rate dissolution that 'triggers' pitting. The implications of these results are that materials processing conditions control the likelihood of corrosion failures, and these data provide a basis for optimizing such conditions.     

Quantum encounters of the cold kind

Since the introduction of laser-cooling techniques for neutral atoms in the early 1980s, the study of collisional interactions between atoms and molecules has been extended to the regime of ultracold temperatures. With nanokelvin temperatures now attainable, our ability to probe the interactions, both experimentally and theoretically, has also progressed. Understanding of the subtle and often highly quantum-mechanical effects that are manifest at such low energies has advanced to the point where new precision measurements are matched by highly accurate theoretical calculations. Low-energy phenomena such as Bose-Einstein condensation and the photoassociation of atoms into bound molecules are now accurately described with no free parameters.     

A nebula of gases from Io surrounding Jupiter

Several planetary missions have reported the presence of substantial numbers of energetic ions and electrons surrounding Jupiter; relativistic electrons are observable up to several astronomical units (au) from the planet. A population of energetic (>30[?]keV) neutral particles also has been reported, but the instrumentation was not able to determine the mass or charge state of the particles, which were subsequently labelled energetic neutral atoms. Although images showing the presence of the trace element sodium were obtained, the source and identity of the neutral atoms---and their overall significance relative to the loss of charged particles from Jupiter's magnetosphere---were unknown. Here we report the discovery by the Cassini spacecraft of a fast (>103[?]km[?]s-1) and hot magnetospheric neutral wind extending more than 0.5[?]au from Jupiter, and the presence of energetic neutral atoms (both hot and cold) that have been accelerated by the electric field in the solar wind. We suggest that these atoms originate in volcanic gases from Io, undergo significant evolution through various electromagnetic interactions, escape Jupiter's magnetosphere and then populate the environment around the planet. Thus a 'nebula' is created that extends outwards over hundreds of jovian radii.