Evidence for “dark charge” from photoluminescence measurements in wide InGaN quantum wells

Wide (15-25 nm) InGaN/GaN quantum wells in LED structures were studied by time-resolved photoluminescence (PL) spectroscopy and compared with narrow (2.6 nm) wells in similar LED structures. Using below-barrier pulsed excitation in the microsecond range, we measured increase and decay of PL pulses. These pulses in wide wells at low-intensity excitation show very slow increase and fast decay. Moreover, the shape of the pulses changes when we vary the separation between them. None of these effects occurs for samples with narrow wells. The unusual properties of wide wells are attributed to the presence of “dark charge” i.e., electrons and holes in the ground states. Their wave functions are spatially separated and due to negligible overlap they do not contribute to emission. However, they screen the built-in field in the well very effectively so that excited states appear with significant overlap and give rise to PL. A simple model of recombination kinetics including “dark charge” explains the observations qualitatively

The dissociation of (a+c) misfit dislocations at the InGaN/GaN interface

(a+c) dislocations in hexagonal materials are typically observed to be dissociated into partial dislocations. Edge (a+c) dislocations are introduced into (0001) nitride semiconductor layers by the process of plastic relaxation. As there is an increasing interest in obtaining relaxed InGaN buffer layers for the deposition of high In content structures, the study of the dissociation mechanism of misfit (a+c) dislocations laying at the InGaN/GaN interface is then crucial for understanding their nucleation and glide mechanisms. In the case of the presented plastically relaxed InGaN layers deposited on GaN substrates we observe a trigonal network of (a+c) dislocations extending at the interface with a rotation of 3 degrees from directions. High resolution microscopy studies show that these dislocations are dissociated into two Frank-Shockley 1/6 partial dislocations with the I1 BSF spreading between them. Atomistic simulations of a dissociated edge (a+c) dislocation revealed a 3/5 atom ring structure for the cores of both partial dislocations. The observed separation between two partial dislocations must result from the climb of at least one of the dislocations during the dissociation process, possibly induced by the mismatch stress in the InGaN layer.Comment: This is a submitted version of the manuscript published in Journal of Microscop

From local watershed management to integrated river basin management at national and transboundary levels

Watersheds face a range of degradation challenges associated with human activities, such as pollution, deforestation and changes in sediment generation. The way they are managed has a profound cascading effect on natural resources and communities in the wider basin. Although watersheds play a critical role as the basic hydrological unit within a river basin they are often neglected in river basin management. Over the past decade, principles and practices have evolved to ensure that integrated water resources management (IWRM) approaches used at the broader basin level to address sustainable development and management of land and water resources also apply at the smaller watershed level. This technical report is a synthesis of the knowledge, lessons learned and good practices presented and discussed at the International Conference on Watershed Management held in Chiang Mai, Thailand 9-11 March 2011

CHMP2B mutants linked to frontotemporal dementia impair maturation of dendritic spines.: CHMP2B and dendritic spines

International audienceThe highly conserved ESCRT-III complex is responsible for deformation and cleavage of membranes during endosomal trafficking and other cellular activities. In humans, dominant mutations in the ESCRT-III subunit CHMP2B cause frontotemporal dementia (FTD). The decade-long process leading to this cortical degeneration is not well understood. One possibility is that, akin to other neurodegenerative diseases, the pathogenic protein affects the integrity of dendritic spines and synapses before any neuronal death. Using confocal microscopy and 3D reconstruction, we examined whether expressing the FTD-linked mutants CHMP2B(intron5) and CHMP2B(Delta10) in cultured hippocampal neurons modified the number or structure of spines. Both mutants induced a significant decrease in the proportion of large spines with mushroom morphology, without overt degeneration. Furthermore, CHMP2B(Delta10) induced a drop in frequency and amplitude of spontaneous excitatory postsynaptic currents, suggesting that the more potent synapses were lost. These effects seemed unrelated to changes in autophagy. Depletion of endogenous CHMP2B by RNAi resulted in morphological changes similar to those induced by mutant CHMP2B, consistent with dominant-negative activity of pathogenic mutants. Thus, CHMP2B is required for spine growth. Taken together, these results demonstrate that a mutant ESCRT-III subunit linked to a human neurodegenerative disease can disrupt the normal pattern of spine development

Structure of cellular ESCRT-III spirals and their relationship to HIV budding

Abstract The ESCRT machinery along with the AAA+ ATPase Vps4 drive membrane scission for trafficking into multivesicular bodies in the endocytic pathway and for the topologically related processes of viral budding and cytokinesis, but how they accomplish this remains unclear. Using deep-etch electron microscopy, we find that endogenous ESCRT-III filaments stabilized by depleting cells of Vps4 create uniform membrane-deforming conical spirals which are assemblies of specific ESCRT-III heteropolymers. To explore functional roles for ESCRT-III filaments, we examine HIV-1 Gag-mediated budding of virus-like particles and find that depleting Vps4 traps ESCRT-III filaments around nascent Gag assemblies. Interpolating between the observed structures suggests a new role for Vps4 in separating ESCRT-III from Gag or other cargo to allow centripetal growth of a neck constricting ESCRT-III spiral. DOI: 10.7554/eLife.02184.00

Rescue of HIV-1 Release by Targeting Widely Divergent NEDD4-Type Ubiquitin Ligases and Isolated Catalytic HECT Domains to Gag

Retroviruses engage the ESCRT pathway through late assembly (L) domains in Gag to promote virus release. HIV-1 uses a PTAP motif as its primary L domain, which interacts with the ESCRT-I component Tsg101. In contrast, certain other retroviruses primarily use PPxY-type L domains, which constitute ligands for NEDD4-type ubiquitin ligases. Surprisingly, although HIV-1 Gag lacks PPxY motifs, the release of HIV-1 L domain mutants is potently enhanced by ectopic NEDD4-2s, a native isoform with a naturally truncated C2 domain that appears to account for the residual titer of L domain-defective HIV-1. The reason for the unique potency of the NEDD4-2s isoform has remained unclear. We now show that the naturally truncated C2 domain of NEDD4-2s functions as an autonomous Gag-targeting module that can be functionally replaced by the unrelated Gag-binding protein cyclophilin A (CypA). The residual C2 domain of NEDD4-2s was sufficient to transfer the ability to stimulate HIV-1 budding to other NEDD4 family members, including the yeast homologue Rsp5, and even to isolated catalytic HECT domains. The isolated catalytic domain of NEDD4-2s also efficiently promoted HIV-1 budding when targeted to Gag via CypA. We conclude that the regions typically required for substrate recognition by HECT ubiquitin ligases are all dispensable to stimulate HIV-1 release, implying that the relevant target for ubiquitination is Gag itself or can be recognized by divergent isolated HECT domains. However, the mere ability to ubiquitinate Gag was not sufficient to stimulate HIV-1 budding. Rather, our results indicate that the synthesis of K63-linked ubiquitin chains is critical for ubiquitin ligase-mediated virus release

Crenarchaeal CdvA Forms Double-Helical Filaments Containing DNA and Interacts with ESCRT-III-Like CdvB

International audienceBACKGROUND: The phylum Crenarchaeota lacks the FtsZ cell division hallmark of bacteria and employs instead Cdv proteins. While CdvB and CdvC are homologues of the eukaryotic ESCRT-III and Vps4 proteins, implicated in membrane fission processes during multivesicular body biogenesis, cytokinesis and budding of some enveloped viruses, little is known about the structure and function of CdvA. Here, we report the biochemical and biophysical characterization of the three Cdv proteins from the hyperthermophilic archaeon Metallospherae sedula. METHODOLOGY/PRINCIPAL FINDINGS: Using sucrose density gradient ultracentrifugation and negative staining electron microscopy, we evidenced for the first time that CdvA forms polymers in association with DNA, similar to known bacterial DNA partitioning proteins. We also observed that, in contrast to full-lengh CdvB that was purified as a monodisperse protein, the C-terminally deleted CdvB construct forms filamentous polymers, a phenomenon previously observed with eukaryotic ESCRT-III proteins. Based on size exclusion chromatography data combined with detection by multi-angle laser light scattering analysis, we demonstrated that CdvC assembles, in a nucleotide-independent way, as homopolymers resembling dodecamers and endowed with ATPase activity in vitro. The interactions between these putative cell division partners were further explored. Thus, besides confirming the previous observations that CdvB interacts with both CdvA and CdvC, our data demonstrate that CdvA/CdvB and CdvC/CdvB interactions are not mutually exclusive. CONCLUSIONS/SIGNIFICANCE: Our data reinforce the concept that Cdv proteins are closely related to the eukaryotic ESCRT-III counterparts and suggest that the organization of the ESCRT-III machinery at the Crenarchaeal cell division septum is organized by CdvA an ancient cytoskeleton protein that might help to coordinate genome segregation