Differences in the Regulation of K-Ras and H-Ras Isoforms by Monoubiquitination

Ras GTPases are signaling switches that control critical cellular processes including gene expression, differentiation, and apoptosis. The major Ras isoforms (K, H, and N) contain a conserved core GTPase domain, but have distinct biological functions. Among the three Ras isoforms there are clear differences in post-translational regulation, which contribute to differences in localization and signaling output. Modification by ubiquitination was recently reported to activate Ras signaling in cells, but the mechanisms of activation are not well understood. Here, we show that H-Ras is activated by monoubiquitination and that ubiquitination at Lys-117 accelerates intrinsic nucleotide exchange, thereby promoting GTP loading. This mechanism of Ras activation is distinct from K-Ras monoubiquitination at Lys-147, which leads to impaired regulator-mediated GTP hydrolysis. These findings reveal that different Ras isoforms are monoubiquitinated at distinct sites, with distinct mechanisms of action, but with a common ability to chronically activate the protein in the absence of a receptor signal or oncogenic mutation.     

M2-like macrophages are responsible for collagen degradation through a mannose receptor–mediated pathway

Tissue remodeling processes critically depend on the timely removal and remodeling of preexisting collagen scaffolds. Nevertheless, many aspects related to the turnover of this abundant extracellular matrix component in vivo are still incompletely understood. We therefore took advantage of recent advances in optical imaging to develop an assay to visualize collagen turnover in situ and identify cell types and molecules involved in this process. Collagen introduced into the dermis of mice underwent cellular endocytosis in a partially matrix metalloproteinase–dependent manner and was subsequently routed to lysosomes for complete degradation. Collagen uptake was predominantly executed by a quantitatively minor population of M2-like macrophages, whereas more abundant Col1a1-expressing fibroblasts and Cx3cr1-expressing macrophages internalized collagen at lower levels. Genetic ablation of the collagen receptors mannose receptor (Mrc1) and urokinase plasminogen activator receptor–associated protein (Endo180 and Mrc2) impaired this intracellular collagen degradation pathway. This study demonstrates the importance of receptor-mediated cellular uptake to collagen turnover in vivo and identifies a key role of M2-like macrophages in this process.     

The Urokinase Receptor Homolog Haldisin Is a Novel Differentiation Marker of Stratum Granulosum in Squamous Epithelia

Several members of the Ly-6/uPAR (LU)-protein domain family are differentially expressed in human squamous epithelia. In some cases, they even play important roles in maintaining skin homeostasis, as exemplified by the secreted single domain member, SLURP-1, the deficiency of which is associated with the development of palmoplantar hyperkeratosis in the congenital skin disorder Mal de Meleda. In the present study, we have characterized a new member of the LU-protein domain family, which we find to be predominantly expressed in the stratum granulosum of human skin, thus resembling the expression of SLURP-1. In accordance with its expression pattern, we denote this protein product, which is encoded by the LYPD5 gene, as Haldisin (human antigen with LU-domains expressed in skin). Two of the five human glycolipid-anchored membrane proteins with multiple LU-domains characterized so far are predominantly confined to squamous epithelia (i.e., C4.4A), to stratum spinosum, and Haldisin to stratum granulosum under normal homeostatic conditions. Whether Haldisin is a prognostic biomarker for certain epithelial malignancies, like C4.4A and SLURP-1, remains to be explored.     

Inhibition of the HIV-1 Spike by Single-PG9/16-Antibody Binding Suggests a Coordinated-Activation Model for Its Three Protomeric Units

The HIV-1 spike is composed of three protomeric units, each containing a peripheral gp120 and a transmembrane gp41 subunit. Binding to the CD4 and the chemokine receptors triggers them to mediate virus entry into cells by membrane fusion. The spikes also represent the major target for neutralizing antibodies (Abs) against the virus. We have studied how two related broadly neutralizing Abs, PG9 and PG16, react with the spike. Unexpectedly, this also suggested how the functions of the individual protomers in the spike depend on each other. The Abs have been shown to bind the V1/V2 loops of gp120, located at the top of the spike. Using blue native-polyacrylamide gel electrophoresis (BN-PAGE), we show that only single Abs or antigen-binding fragments could bind to the spikes of HIV-1 virus-like particles. Apparently, binding to one gp120 sterically interferes with binding to the other two subunits in the spike top. Despite this constraint, all of the protomers of the spike became resistant to CD4 binding and subsequent formation of the coreceptor binding site. These activities were measured by monitoring the sequential complex formation of the spike first with Abs and then with soluble 2d- or 4d-CD4 or with soluble CD4 and the CD4 inducible coreceptor binding site Ab 17b in BN-PAGE. The inhibition of the spike by single-Ab binding suggested that the activation reactions of the individual protomeric units are linked to each other in a coordinated activation process.     

Quotas on Narwhal (Monodon monoceros) Hunting in East Greenland: Trends in Narwhal Killed per Hunter and Potential Impacts of Regulations on Inuit Communities

This study evaluates the introduction of quotas on narwhal hunting in East Greenland with respect to effects on Inuit culture and based on trends in narwhal killed per hunter and assessment of migration patterns. Cultural aspects were assessed through group discussions and comparison between East and Northwest Greenland. Trends in narwhal killed/hunter were modeled from catch statistics using information on number of hunters and climate and ice cover data for the period 1993–2004. Results indicate negative impacts of quotas on Inuit culture; did not detect negative trends in narwhal killed/hunter; and suggest south-west-bound migration, implying potential immigration from non-hunted populations that was not considered in quota setting. The implementation of quotas without local consultations and legal basis in the relevant executive order is therefore in our opinion inappropriate. Conservation and sustainable use of narwhal stocks may be more likely to succeed if local communities are involved through co-management agreements.     

Denning behaviour of ship rats (Rattus rattus) on Taukihepa,a seabird breeding island

Abstract

Den sites of 14 ship rats (Rattus rattus) were located daily during the rat breeding season on Taukihepa (Big South Cape), a seabird island southwest of Rakiura (Stewart Island). In contrast to other New Zealand studies, no arboreal dens were found. Den sites on Taukihepa were in ferns, under logs, in woodpiles, or underground in sooty shearwater (Puffinusgriseus) breeding burrows. The number of times known den sites used was positively related to the amount of leaf litter and woodpiles near the den sites. Overall, 24% of radio‐tagged rats were sharing den sites on any given day. While there was considerable individual variation in the number of times den sites were used, female rats tended to reuse den sites more than males. Many rats were found in dens alone, but frequently males and females shared. Occasionally two females and one male denned together, as did two females, whereas males never denned with another male.     

Disturbance and Resilience in Tropical American Palm Populations and Communities

We review resilience to natural and anthropogenic disturbance of palm populations and communities in tropical America. Response of palms to disturbance depends on their morphological traits, their reproductive strategies and the impacts of these traits and strategies on phenology and gene flow. Human impact induces changes in genetic structure, increasing endogamy and genetic drift in fragmented populations. Forest fragmentation and harvest of palm organs are well documented whereas effects of intermediate disturbance like selective logging, hunting or fire remain poorly known. We recommend emphasis on long-term experiments and on the use of mechanistic approaches in future research to facilitate integration of available data into a theoretical ecological framework.     

Palm Management in South America

We reviewed information on management of useful palms in South America. We documented management for 96 species, from incidental activities intended to increase populations of wild palms to the inclusion of palms in complex agroforestry systems. Two species, Bactris gasipaes and Parajubaea cocoides, are domesticated. Managed species are remarkably fewer than species used in the region, which suggests that harvesters often disregard the fate of the species they use. The best way of managing palms is to employ harvest methods that do not decimate the populations. Although a variety of harvesting techniques have been documented, overharvest is common, and mismanagement prevails – unnecessary felling of palms in order to harvest leaves or fruits is a widespread practice. Research should focus on assessing production in response to management practices, but eradicating the habit of destructive harvest is an obvious priority. Research on palm management must be combined with actions addressed to all stakeholders of the palm/humans system.