Can Individual and Social Patterns of Resource Use Buffer Animal Populations against Resource Decline?

Species in many ecosystems are facing declines of key resources. If we are to understand and predict the effects of resource loss on natural populations, we need to understand whether and how the way animals use resources changes under resource decline. We investigated how the abundance of arboreal marsupials varies in response to a critical resource, hollow-bearing trees. Principally, we asked what mechanisms mediate the relationship between resources and abundance? Do animals use a greater or smaller proportion of the remaining resource, and is there a change in cooperative resource use (den sharing), as the availability of hollow trees declines? Analyses of data from 160 sites surveyed from 1997 to 2007 showed that hollow tree availability was positively associated with abundance of the mountain brushtail possum, the agile antechinus and the greater glider. The abundance of Leadbeater’s possum was primarily influenced by forest age. Notably, the relationship between abundance and hollow tree availability was significantly less than 1∶1 for all species. This was due primarily to a significant increase by all species in the proportional use of hollow-bearing trees where the abundance of this resource was low. The resource-sharing response was weaker and inconsistent among species. Two species, the mountain brushtail possum and the agile antechinus, showed significant but contrasting relationships between the number of animals per occupied tree and hollow tree abundance. The discrepancies between the species can be explained partly by differences in several aspects of the species’ biology, including body size, types of hollows used and social behaviour as it relates to hollow use. Our results show that individual and social aspects of resource use are not always static in response to resource availability and support the need to account for dynamic resource use patterns in predictive models of animal distribution and abundance.     

Planted Riparian Buffer Zones in New Zealand: Do They Live Up to Expectations?

Abstract River and stream rehabilitation projects are increasing in number, but the success or failure of these projects has rarely been evaluated, and the extent to which buffers can restore riparian and stream function and species composition is not well understood. In New Zealand the widespread conversion of forest to agricultural land has caused degradation of streams and riparian ecosystems. We assessed nine riparian buffer zone schemes in North Island, New Zealand that had been fenced and planted (age range from 2 to 24 years) and compared them with unbuffered control reaches upstream or nearby. Macroinvertebrate community composition was our prime indicator of water and habitat quality and ecological functioning, but we also assessed a range of physical and water quality variables within the stream and in the riparian zone. Generally, streams within buffer zones showed rapid improvements in visual water clarity and channel stability, but nutrient and fecal contamination responses were variable. Significant changes in macroinvertebrate communities toward “clean water” or native forest communities did not occur at most of the study sites. Improvement in invertebrate communities appeared to be most strongly linked to decreases in water temperature, suggesting that restoration of in‐stream communities would only be achieved after canopy closure, with long buffer lengths, and protection of headwater tributaries. Expectations of riparian restoration efforts should be tempered by (1) time scales and (2) spatial arrangement of planted reaches, either within a catchment or with consideration of their proximity to source areas of recolonists.     

“cAMP Sponge”: A Buffer for Cyclic Adenosine 3′, 5′-Monophosphate

Background

While intracellular buffers are widely used to study calcium signaling, no such tool exists for the other major second messenger, cyclic AMP (cAMP).

Methods/Principal Findings

Here we describe a genetically encoded buffer for cAMP based on the high-affinity cAMP-binding carboxy-terminus of the regulatory subunit RIβ of protein kinase A (PKA). Addition of targeting sequences permitted localization of this fragment to the extra-nuclear compartment, while tagging with mCherry allowed quantification of its expression at the single cell level. This construct (named “cAMP sponge”) was shown to selectively bind cAMP in vitro. Its expression significantly suppressed agonist-induced cAMP signals and the downstream activation of PKA within the cytosol as measured by FRET-based sensors in single living cells. Point mutations in the cAMP-binding domains of the construct rendered the chimera unable to bind cAMP in vitro or in situ. Cyclic AMP sponge was fruitfully applied to examine feedback regulation of gap junction-mediated transfer of cAMP in epithelial cell couplets.

Conclusions

This newest member of the cAMP toolbox has the potential to reveal unique biological functions of cAMP, including insight into the functional significance of compartmentalized signaling events.     

Double trouble—Buffer selection and His-tag presence may be responsible for nonreproducibility of biomedical experiments

The availability of purified and active protein is the starting point for the majority of in vitro biomedical, biochemical, and drug discovery experiments. The use of polyhistidine affinity tags has resulted in great increases of the efficiency of the protein purification process, but can negatively affect structure and/or activity measurements. Similarly, buffer molecules may perturb the conformational stability of a protein or its activity. During the determination of the structure of a Gcn5-related N-acetyltransferase (GNAT) from Pseudomonas aeruginosa (PA4794), we found that both HEPES and the polyhistidine affinity tag bind (separately) in the substrate-binding site. In the case of HEPES, the molecule induces conformational changes in the active site, but does not significantly affect enzyme activity. In contrast, the uncleaved His-tag does not induce major conformational changes but acts as a weak competitive inhibitor of peptide substrate. In two other GNAT enzymes, we observed that the presence of the His-tag had a strong influence on the activity of these proteins. The influence of protein preparation on functional studies may affect the reproducibility of experiments in other laboratories, even when changes between protocols seem at first glance to be insignificant. Moreover, the results presented here show how critical it is to adjust the experimental conditions for each protein or family of proteins, and investigate the influence of these factors on protein activity and structure, as they may significantly alter the effectiveness of functional characterization and screening methods. Thus, we show that a polyhistidine tag and the buffer molecule HEPES bind in the substrate-binding site and influence the conformation of the active site and the activity of GNAT acetyltransferases. We believe that such discrepancies can influence the reproducibility of some experiments and therefore could have a significant “ripple effect” on subsequent studies.     

Can a reductionist be a pluralist?

Pluralism is often put forth as a counter-position to reductionism. In this essay, I argue that reductionism and pluralism are in fact consistent. I propose that there are several potential goals for reductions and that the proper form of a reduction should be considered in tandem with the goal that it aims to achieve. This insight provides a basis for clarifying what version(s) of reductionism are currently defended, for explicating the notion of a fundamental level of explanation, and for showing how one can be both a reductionist and a pluralist.     

When is a peroxisome not a peroxisome?

It is time to drop the glyoxysome name. Recent functional genomics analysis together with cell biology studies emphasize the unifying features of peroxisomes rather than their differences. Plant peroxisomes contain 300 or more proteins, the functions of which are dominated by activities related to fatty acid oxidation (>70 enzymes). By comparison, relatively few proteins are committed to metabolism of reactive oxygen species ( approximately 20) and to photorespiration ( approximately 10). Analysis of triglyceride metabolism in Arabidopsis seedlings now indicates that only two enzymes (isocitrate lyase and malate synthase) potentially distinguish glyoxysomes from other peroxisomes. Future research is best served by focusing on the common features of peroxisomes to establish how these dynamic organelles contribute to energy metabolism, development and responses to environmental challenges.     

Between a rock and a hard place?

Developing small-molecule inhibitors against protein-protein interaction targets is among the most difficult challenges in contemporary drug discovery. Recent developments in our understanding of this problem, and in the knowledge and tools available to address it, give cause for renewed hope, but substantial challenges remain.     

When is a parasite species a species?

Regrettably, 140 years after the publication of Darwin's Origin of Species, we face the grotesque situation that we still do not know what is a species whose origin Darwin wanted to explain. A generally applicable species definition is not available. Is there a basic unit of biodiversity above the level of individuals? Do we try to define something that does not exist in reality? The strong potential for the evolution of genetic variability in parasites together with the importance of species diagnosis for applied fields of parasite research make biodiversity research a key role in parasitology. Frequent occurrence of sympatric speciation, clonal reproduction, selfing, sib mating or parthenogenesis imply exceptional conditions for the evolution of gene pool diversities in parasites.     

When is a herbivore not a herbivore?

Summary Young herbivores are not herbivores. With the apparent exception of some chewing insects they cannot survive and growth without eating animal or microbial protein. Further reinforcing how short available nitrogen is in plant food, adult herbivores must feed selectively and depend on intestinal microorganisms to obtain sufficient nitrogen for maintenance and breeding.     

What Makes a Protein Sequence a Prion?

Typical amyloid diseases such as Alzheimer''s and Parkinson''s were thought to exclusively result from de novo aggregation, but recently it was shown that amyloids formed in one cell can cross-seed aggregation in other cells, following a prion-like mechanism. Despite the large experimental effort devoted to understanding the phenomenon of prion transmissibility, it is still poorly understood how this property is encoded in the primary sequence. In many cases, prion structural conversion is driven by the presence of relatively large glutamine/asparagine (Q/N) enriched segments. Several studies suggest that it is the amino acid composition of these regions rather than their specific sequence that accounts for their priogenicity. However, our analysis indicates that it is instead the presence and potency of specific short amyloid-prone sequences that occur within intrinsically disordered Q/N-rich regions that determine their prion behaviour, modulated by the structural and compositional context. This provides a basis for the accurate identification and evaluation of prion candidate sequences in proteomes in the context of a unified framework for amyloid formation and prion propagation.     

Is there a pilot in a pseudopod?

The concept of pilot pseudopodia is reconsidered 30 years after its inauguration (Gerisch, G., Hülser, D., Malchow, D., Wick, U., 1975. Cell communication by periodic cyclic-AMP pulses. Phil. Trans. R. Soc. Lond. B 272, 181-192). The original hypothesis stated that protruding pseudopodia serve as dynamic sensory organelles that aid a cell in perceiving variations of chemoattractant concentration and, consequently, in navigation during chemotaxis. This influential idea is reevaluated in the light of recent findings about the mechanisms governing chemotactic cell motility, morphology and dynamics of pseudopodia, and about molecular constituents and regulators of pseudopod extension and retraction. It is proposed that stimulation by a chemoattractant modulates speed of pseudopod protrusion and thereby increases cell elongation. Elongation further enhances chemotactic sensitivity of the cell to shallow chemoattractant gradients, reinforces cell polarization, and finally leads to suppression of lateral pseudopodia and continuation of cell migration in the gradient direction.