Crystallization of SHARPIN using an automated two-dimensional grid screen for optimization

An N-terminal fragment of human SHARPIN was recombinantly expressed in Escherichia coli, purified and crystallized. Crystals suitable for X-ray diffraction were obtained by a one-step optimization of seed dilution and protein concentration using a two-dimensional grid screen. The crystals belonged to the primitive tetragonal space group P4(3)2(1)2, with unit-cell parameters a = b = 61.55, c = 222.81 Å. Complete data sets were collected from native and selenomethionine-substituted protein crystals at 100 K to 2.6 and 2.0 Å resolution, respectively

Possibilities offered by implantable miniaturized cuff-electrodes for insect neurophysiology

AbstractRecent advances in microsystems technology led to a miniaturization of cuff-electrodes, which suggests these electrodes not just for long-term neuronal recordings in mammalians, but also in medium-sized insects. In this study we investigated the possibilities offered by cuff-electrodes for neuroethology using insects as a model organism. The implantation in the neck of a tropical bushcricket resulted in high quality extracellular nerve recordings of different units responding to various acoustic, vibratory, optical and mechanical stimuli. In addition, multi-unit nerve activity related to leg movements was recorded in insects walking on a trackball. A drawback of bi-polar nerve recordings obtained during tethered flight was overlay of nerve activity with large amplitude muscle potentials. Interestingly, cuff-electrode recordings were robust to withstand walking and flight activity so that good quality nerve recordings were possible even three days after electrode implantation. Recording multi-unit nerve activity in intact insects required an elaborate spike sorting algorithm in order to discriminate neuronal units responding to external stimuli from background activity. In future, a combination of miniaturized cuff-electrodes and light-weight amplifiers equipped with a wireless transmitter will allow the investigation of neuronal processes underlying natural behavior in freely moving insects. By this means cuff-electrodes may contribute to the development of realistic neuronal models simulating neuronal processes underlying natural insect behavior, such like mate choice and predator avoidance

Gulf Stream density structure and transport during the past millennium

The Gulf Stream transports approximately 31 Sv (1 Sv = 10^6 m^3 s^(-1)) of water and 1.3 10^(15) W of heat into the North Atlantic ocean. The possibility of abrupt changes in Gulf Stream heat transport is one of the key uncertainties in predictions of climate change for the coming centuries. Given the limited length of the instrumental record, our knowledge of Gulf Stream behaviour on long timescales must rely heavily on information from geologic archives. Here we use foraminifera from a suite of high-resolution sediment cores in the Florida Straits to show that the cross-current density gradient and vertical current shear of the Gulf Stream were systematically lower during the Little Ice Age (AD ~1200 to 1850). We also estimate that Little Ice Age volume transport was ten per cent weaker than today's. The timing of reduced flow is consistent with temperature minima in several palaeoclimate records, implying that diminished oceanic heat transport may have contributed to Little Ice Age cooling in the North Atlantic. The interval of low flow also coincides with anomalously high Gulf Stream surface salinity, suggesting a tight linkage between the Atlantic Ocean circulation and hydrologic cycle during the past millennium

LUBAC synthesizes linear ubiquitin chains via a thioester intermediate

The linear ubiquitin chain assembly complex (LUBAC) is a RING E3 ligase that regulates immune and inflammatory signalling pathways. Unlike classical RING E3 ligases, LUBAC determines the type of ubiquitin chain being formed, an activity normally associated with the E2 enzyme. We show that the RING-in-between-RING (RBR)-containing region of HOIP—the catalytic subunit of LUBAC—is sufficient to generate linear ubiquitin chains. However, this activity is inhibited by the N-terminal portion of the molecule, an inhibition that is released upon complex formation with HOIL-1L or SHARPIN. Furthermore, we demonstrate that HOIP transfers ubiquitin to the substrate through a thioester intermediate formed by a conserved cysteine in the RING2 domain, supporting the notion that RBR ligases act as RING/HECT hybrids

Deglacial variability in the surface return flow of the Atlantic meridional overturning circulation

Author Posting. © American Geophysical Union, 2008. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Paleoceanography 23 (2008): PA1217, doi:10.1029/2007PA001450.Benthic foraminiferal Cd/Ca from a Florida Current sediment core documents the history of the northward penetration of southern source waters within the surface return flow of the Atlantic meridional overturning circulation (AMOC). Cd seawater estimates (CdW) indicate that intermediate-depth southern source waters crossed the equator and contributed to the Florida Current during the Bølling-Allerød warm period of the last deglaciation, consistent with evidence of only a modest AMOC reduction compared to today. The CdW estimates also provide the first paleoceanographic evidence of a reduction in the influence of intermediate-depth southern source waters within the Florida Current during the Younger Dryas, a deglacial cold event characterized by a weak North Atlantic AMOC. Our results reveal a close correspondence between the northward penetration of intermediate-depth southern source waters and the influence of North Atlantic Deep Water, suggesting a possible link between intermediate-depth southern source waters and the strength of the Atlantic AMOC.This work was funded by the NSF and the WHOI Ocean and Climate Change Institute

Evidence from the Florida Straits for Younger Dryas ocean circulation changes

Author Posting. © American Geophysical Union, 2011. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Paleoceanography 26 (2011): PA1205, doi:10.1029/2010PA002032.The waters passing through the Florida Straits today reflect both the western portion of the wind-driven subtropical gyre and the northward flow of the upper waters which cross the equator, compensating North Atlantic Deep Water export as part of the large-scale Atlantic meridional overturning circulation. It has been postulated from various lines of evidence that the overturning circulation was weaker during the Younger Dryas cold event of the last deglaciation. We show here that the contrast in the oxygen isotopic composition of benthic foraminiferal tests across the Florida Current is reduced during the Younger Dryas. This most likely reflects a decrease in the density gradient across the channel and a decrease in the vertical shear of the Florida Current. This reduced shear is consistent with the postulated reduction in the Atlantic meridional overturning circulation. We find that the onset of this change in density structure and flow at the start of the Younger Dryas is very abrupt, occurring in less than 70 years.We thank the National Science Foundation (grants OCE‐0648258 and OCE‐0096472) and the Comer Science and Education Foundation for supporting this research. MWS was supported by a NOAA Global Change Postdoctoral Fellowship