Proteomic and microarray analyses of the Dictyostelium Zak1-GSK-3 signaling pathway reveal a role in early development

GskA, the Dictyostelium GSK-3 orthologue, is modified and activated by the dual-specificity tyrosine kinase Zak1, and the two kinases form part of a signaling pathway that responds to extracellular cyclic AMP. We identify potential cellular effectors for the two kinases by analyzing the corresponding null mutants. There are proteins and mRNAs that are altered in abundance in only one or the other of the two mutants, indicating that each kinase has some unique functions. However, proteomic and microarray analyses identified a number of proteins and genes, respectively, that are similarly misregulated in both mutant strains. The positive correlation between the array data and the proteomic data is consistent with the Zak1-GskA signaling pathway's functioning by directly or indirectly regulating gene expression. The discoidin 1 genes are positively regulated by the pathway, while the abundance of the H5 protein is negatively regulated. Two of the targets, H5 and discoidin 1, are well-characterized markers for early development, indicating that the Zak1-GskA pathway plays a role in development earlier than previously observed.     

The ligand Jelly Belly (Jeb) activates the Drosophila Alk RTK to drive PC12 cell differentiation, but is unable to activate the mouse ALK RTK

The Drosophila Alk receptor tyrosine kinase (RTK) drives founder cell specification in the developing visceral mesoderm and is crucial for the formation of the fly gut. Activation of Alk occurs in response to the secreted ligand Jelly Belly. No homologues of Jelly Belly are described in vertebrates, therefore we have approached the question of the evolutionary conservation of the Jeb-Alk interaction by asking whether vertebrate ALK is able to function in Drosophila. Here we show that the mouse ALK RTK is unable to rescue a Drosophila Alk mutant, indicating that mouse ALK is unable to recognise and respond to the Drosophila Jeb molecule. Furthermore, the overexpression of a dominant-negative Drosophila Alk transgene is able to block the visceral muscle fusion event, which an identically designed dominant-negative construct for the mouse ALK is not. Using PC12 cells as a model for neurite outgrowth, we show here for the first time that activation of dAlk by Jeb results in neurite extension. However, the mouse Alk receptor is unable to respond in any way to the Drosophila Jeb protein in the PC12 system. In conclusion, we find that the mammalian ALK receptor is unable to respond to the Jeb ligand in vivo or in vitro. These results suggest that either (i) mouse ALK and "mouse Jeb" have co-evolved to the extent that mALK can no longer recognise the Drosophila Jeb ligand or (ii) that the mALK RTK has evolved such that it is no longer activated by a Jeb-like molecule in vertebrates.     

The first description of the potentially toxic dinoflagellate, Alexandrium minutum in Hunts Bay, Kingston Harbour, Jamaica

The occurrence and morphology of the potentially toxic dinoflagellate species Alexandrium minutum found for the first time in Jamaica, were examined and described by light and scanning electron microscopy. Classical morphological examinations of whole cells, the thecal plate pattern of intact cells and more importantly the structure of individual thecal plates of squashed cells, were conducted in an attempt to positively identify the species. Characteristics such as a tear-drop shaped apical pore plate with a comma-shaped apical pore and no anterior attachment pore; a narrow sixth precingular plate; a narrow anterior sulcal plate longer than or approximately as long as it is wide; and a posterior sulcal plate wider than long, confirmed the Jamaican species as A. minutum. This dinoflagellate which produces potent neurotoxins responsible for paralytic shellfish poisoning (PSP) in humans in many parts of the World, as well as mass mortality of various marine flora and fauna, was identified in water samples collected during an extensive bloom of the species in the brackish to saline water body of Hunts Bay, an estuarine arm of Kingston Harbour, Jamaica in August 1994. The highest cell concentration was 4.6 × 10⁵ cells l⁻¹, a concentration which far exceeds acceptable concentrations (<10³ cells l⁻¹) of PSP-toxin producing A. minutum in several countries including: Spain and Denmark. No PSP human symptoms were reported during the bloom; however it was accompanied by a large kill of small pelagic fish extending across a third of the bay. Since then, smaller blooms of A. minutum have occurred with the most recent in February and April 2004. Hunts Bay is an important fishing, shrimping and to some extent oyster/mussel collection area and provides an important source of livelihood and food for many fishermen in nearby fishing communities as well as an important source of food for members of other communities. Although there are no known records of human illness due to PSP in Jamaica, the occurrence and blooming in Jamaican waters of this potentially toxic dinoflagellate, is great cause for concern.     

Correction to: Managing biological invasions: the cost of inaction

Biological Invasions -     

Introduction pathways of economically costly invasive alien species

Biological Invasions - Introduction pathways play a pivotal role in the success of Invasive Alien Species (IAS)—the subset of alien species that have a negative environmental and/or...     

Evolution of cranial shape in a continental‐scale evolutionary radiation of Australian lizards

A defining character of adaptive radiations is the evolution of a diversity of morphological forms that are associated with the use of different habitats, following the invasion of vacant niches. Island adaptive radiations have been thoroughly investigated but continental scale radiations are more poorly understood. Here, we use 52 species of Australian agamid lizards and their Asian relatives as a model group, and employ three‐dimensional geometric morphometrics to characterize cranial morphology and investigate whether variation in cranial shape reflects patterns expected from the ecological process of adaptive radiation. Phylogenetic affinity, evolutionary allometry, and ecological life habit all play major roles in the evolution of cranial shape in the sampled lizards. We find a significant association between cranial shapes and life habit. Our results are in line with the expectations of an adaptive radiation, and this is the first time detailed geometric morphometric analyses have been used to understand the selective forces that drove an adaptive radiation at a continental scale.     

Non-native species and lake warming negatively affect Arctic char Salvelinus alpinus abundance; deep thermal refugia facilitate co-existence

This study finds that non-native species and warming temperatures have significant negative effects on Arctic char Salvelinus alpinus abundance in Irish lakes. Eutrophication was not important at the range of total phosphorus tested (0.005–0.023 mg l⁻¹). Model results predict that S. alpinus occur across the temperature range sampled (8.2–19.7°C) when non-natives are absent, but S. alpinus catch is predicted to be close to zero irrespective of temperature when non-native catch is high. This result indicates that to persist, S. alpinus may require a habitat where non-natives are at low abundance or absent. Salvelinus alpinus segregated from other species along the thermal axis, inhabiting significantly colder water and actively avoided non-native species, which appeared to limit their distribution. The thermal niche realized by S. alpinus in non-native dominated lakes was thus compressed relative to native dominated lakes and S. alpinus population density was significantly lower. These findings were consistent even when the only non-native present was Perca fluviatilis. Temperature appeared to limit the distribution of non-native species, such that the presence of deep thermal refugia is currently facilitating S. alpinus co-existence with non-natives in associated lakes. Diet analysis identified P. fluviatilis as potential predators and competitors. This study provides strong evidence that non-native species are a key driver of low S. alpinus abundance in Irish lakes. Temperature increases associated with climate change are identified as a secondary concern, as they could erode S. alpinus' thermal niche and lead to their extirpation. The lower thermal buffering capacity of shallow lakes identifies these as higher risk systems. Salvelinus alpinus conservation in Ireland should focus on preventing future illegal non-native species introductions because unlike other stressors (e.g., eutrophication etc.), species introductions are rarely reversible.     

Escherichia coli “TatExpress” strains export several g/L human growth hormone to the periplasm by the Tat pathway

Escherichia coli is a heavily used platform for the production of biotherapeutic and other high-value proteins, and a favored strategy is to export the protein of interest to the periplasm to simplify downstream processing and facilitate disulfide bond formation. The Sec pathway is the standard means of transporting the target protein but it is unable to transport complex or rapidly folding proteins because the Sec system can only transport proteins in an unfolded state. The Tat system also operates to transport proteins to the periplasm, and it has significant potential as an alternative means of recombinant protein production because it transports fully folded proteins. Here, we have tested the Tat system's full potential for the production of biotherapeutics for the first time using fed-batch fermentation. We expressed human growth hormone (hGH) with a Tat signal peptide in E. coli W3110 “TatExpress” strains that contain elevated levels of the Tat apparatus. This construct contained four amino acids from TorA at the hGH N-terminus as well as the initiation methionine from hGH, which is removed in vivo. We show that the protein is efficiently exported to the periplasm during extended fed-batch fermentation, to the extent that it is by far the most abundant protein in the periplasm. The protein was shown to be homogeneous, disulfide bonded, and active. The bioassay showed that the yields of purified periplasmic hGH are 5.4 g/L culture whereas an enzyme-linked immunosorbent assay gave a figure of 2.39 g/L. Separate analysis of a TorA signal peptide linked to hGH construct lacking any additional amino acids likewise showed efficient export to the periplasm, although yields were approximately two-fold lower.     

Laser chasing behaviour of wild fishes exploited as a tool to compare space use between size,sex and species

The spatial extent of animal movement is a key consideration when designing conservation measures, such as marine protected areas. Methods to assess territory size in the marine environment, however, are labour intensive and/or expensive. Here, we explore a novel method to investigate the spatial ecology of territorial fishes by examining their reactions to an artificial light stimulus. During benthic towed video surveys conducted in Lyme Bay, southwest England, several species of wrasse (Labridae) have frequently been observed pursuing a laser projected onto the seabed. While the motivation behind ‘laser‐chasing’ is unclear, we quantified the spatial aspects of this behaviour by comparing chase distance and chase likelihood between and within species, to determine the potential utility of this method for investigating space use and aggression in wild fishes. Cuckoo wrasse (Labrus mixtus) were significantly more likely to display agonistic behaviour towards the laser than Goldsinny wrasse (Ctenolabrus rupestris). Goldsinny wrasse displayed a positive relationship between total length and chase‐distance, but not Cuckoo wrasse. The observed species differences may relate to behavioural factors affecting the motivation behind ‘laser‐chasing’, which is discussed. Chases by the cuckoo wrasse were significantly longer than those by Goldsinny wrasse, and these chase distances were used to estimate theoretical territory sizes for each species. To our knowledge, this is the first study to explore the spatial aspects of the reactions to an artificial stimulus by wild fishes. The potential to develop the method to directly investigate aspects of territoriality and aggression in wild fishes is discussed, including necessary further refinements and testing. Wild wrasses are increasingly exploited in Europe to provide cleaner fish for salmonid aquaculture, and we encourage the development of methods to inform spatial conservation measures for these ubiquitous and iconic species.