Human MRCKα is regulated by cellular iron levels and interferes with transferrin iron uptake

Myotonic dystrophy kinase-related Cdc42-binding kinase α (MRCKα, formally known as CDC42BPA) is a serine/threonine kinase that can regulate actin/myosin assembly and activity. Recently, it has been shown that it possesses a functional iron responsive element (IRE) in the 3′-untranslated region (UTR) of its mRNA, suggesting that it may be involved in iron metabolism. Here we report that MRCKα protein expression is also regulated by iron levels; MRCKα colocalizes with transferrin (Tf)-loaded transferrin receptors (TfR), and attenuation of MRCKα expression by a short hairpin RNA silencing construct leads to a significant decrease in Tf-mediated iron uptake. Our results thus indicate that MRCKα takes part in Tf-iron uptake, probably via regulation of Tf-TfR endocytosis/endosome trafficking that is dependent on the cellular cytoskeleton. Regulation of the MRCKα activity by intracellular iron levels could thus represent another molecular feedback mechanism cells could use to finely tune iron uptake to actual needs.     

Spontaneous succession in limestone quarries as an effective restoration tool for endangered arthropods and plants

1. The view of post-mining sites is rapidly changing among ecologists and conservationists, as sensitive restoration using spontaneous succession may turn such sites into biodiversity refuges in human-exploited regions. However, technical reclamation, consisting of covering the sites by topsoil, sowing fast-growing herb mixtures and planting trees, is still commonly adopted. Until now, no multi-taxa study has compared technically reclaimed sites and sites left with spontaneous succession.
2. We sampled communities of vascular plants and 10 arthropod groups in technically reclaimed and spontaneously restored plots in limestone quarries in the Bohemian Karst, Czech Republic. For comparison, we used paired t -tests and multivariate methods, emphasizing red-list status and habitat specialization of individual species.
3. We recorded 692 species of target taxa, with a high proportion of red-listed (10%) and xeric specialist (14%) species, corroborating the great conservation potential of the quarries.
4. Spontaneously restored post-mining sites did not differ in species richness from the technical reclaimed sites but they supported more rare species. The microhabitat cover of leaf litter, herbs and moss, were all directly influenced by the addition of topsoil during reclamation.
5. Synthesis and applications. Our results show that the high conservation potential of limestone quarries could be realized by allowing succession to progress spontaneously with minimal intervention. Given the threat to semi-natural sparsely vegetated habitats in many regions, active restoration measures at post-mining sites should be limited to maintenance of early successional stages, instead of acceleration of succession.     

CAIcal: A combined set of tools to assess codon usage adaptation

 

A recent systematic survey suggested that the YRG (or YawG/YlqF) family with the G4-G5-G1-G2-G3 order of the conserved GTPase motifs represents the only possible circularly permuted variation of the canonical GTPase structure. Here we show that a different circularly permuted GTPase domain actually does exist, conforming to the pattern G3-G4-G5-G1-G2. The domain, dubbed cpRAS, is a variant of RAS family GTPases and occurs in two types of larger proteins, either inserted into a region homologous to a bacterial group of proteins classified as COG2373 and potentially related to the alpha-2-macroglobulin family (so far a single protein in Dictyostelium) or in combination with a von Willebrand factor type A (VWA) domain. For the latter protein type, which was found in a few metazoans and several distantly related protists, existence in the common ancestor of opisthokonts, Amoebozoa and excavates followed by at least eight independent losses may be inferred. Our findings thus bring further evidence for the importance of parallel reduction of ancestral complexity in the eukaryotic evolution.

Reviewers

This article was reviewed by Lakshminarayan Iyer and Fyodor Kondrashov. For the full reviews, please go to the Reviewers' comments section.     

Frataxin, a conserved mitochondrial protein, in the hydrogenosome of Trichomonas vaginalis

Recent data suggest that frataxin plays a key role in eukaryote cellular iron metabolism, particularly in mitochondrial heme and iron-sulfur (FeS) cluster biosynthesis. We have now identified a frataxin homologue (T. vaginalis frataxin) from the human parasite Trichomonas vaginalis. Instead of mitochondria, this unicellular eukaryote possesses hydrogenosomes, peculiar organelles that produce hydrogen but nevertheless share common ancestry with mitochondria. T. vaginalis frataxin contains conserved residues implicated in iron binding, and in silico, it is predicted to form a typical alpha-beta sandwich motif. The short N-terminal extension of T. vaginalis frataxin resembles presequences that target proteins to hydrogenosomes, a prediction confirmed by the results of overexpression of T. vaginalis frataxin in T. vaginalis. When expressed in the mitochondria of a frataxin-deficient Saccharomyces cerevisiae strain, T. vaginalis frataxin partially restored defects in heme and FeS cluster biosynthesis. Although components of heme synthesis or heme-containing proteins have not been found in T. vaginalis to date, T. vaginalis frataxin was also shown to interact with S. cerevisiae ferrochelatase by using a Biacore assay. The discovery of conserved iron-metabolizing pathways in mitochondria and hydrogenosomes provides additional evidence not only of their common evolutionary history, but also of the fundamental importance of this pathway for eukaryotes.     

Fatty acid modification of Wnt1 and Wnt3a at serine is prerequisite for lipidation at cysteine and is essential for Wnt signalling

The Wnt family of proteins is a group of extracellular signalling molecules that regulate cell-fate decisions in developing and adult tissues. It is presumed that all 19 mammalian Wnt family members contain two types of post-translational modification: the covalent attachment of fatty acids at two distinct positions, and the N-glycosylation of multiple asparagines. We examined how these modifications contribute to the secretion, extracellular movement and signalling activity of mouse Wnt1 and Wnt3a ligands. We revealed that O-linked acylation of serine is required for the subsequent S-palmitoylation of cysteine. As such, mutant proteins that lack the crucial serine residue are not lipidated. Interestingly, although double-acylation of Wnt1 was indispensable for signalling in mammalian cells, in Xenopus embryos the S-palmitoyl-deficient form retained the signalling activity. In the case of Wnt3a, the functional duality of the attached acyls was less prominent, since the ligand lacking S-linked palmitate was still capable of signalling in various cellular contexts. Finally, we show that the signalling competency of both Wnt1 and Wnt3a is related to their ability to associate with the extracellular matrix.     

Role of FAT/CD36 in novel PKC isoform activation in heart of spontaneously hypertensive rats

Disruption to the sensitive balance of long-chain fatty acids and glucose in the heart could cause cardiovascular diseases. Searching for a possible role of novel protein kinase C (nPKC) in heart with disrupted energy balance, we compared the insulin-resistant spontaneously hypertensive rats (SHR), which carry a nonfunctional variant of the fatty acid transporter FAT/CD36, with the less insulin-resistant congenic strain SHR-4 that is genetically identical except for a segment on chromosome 4 including a wild-type gene for a functional FAT/CD36. We analyzed expression of the nPKC-δ and -ε isoforms plus triacylglycerols (TAG) content in the myocardium of both FAT/CD36 strains and after a high sucrose diet (HSD). Two weeks before killing, males of both strains were randomly divided into two groups and fed either a standard laboratory chow or an HSD. PKC was determined by Western blotting in particulate and cytosolic fractions from left ventricles. The SHR-4 rats exhibited lower serum levels of insulin and free fatty acids than did SHR rats and higher amounts of PKC-ε in the heart particulate fraction. HSD caused accumulation of heart TAG in SHR but not in SHR-4. HSD increased PKC-δ and decreased PKC-ε expression in particulate fraction from left ventricles of SHR-4 while having no effects in SHR. These results demonstrate that reduced insulin resistance in SHR-4 rats with wild-type FAT/CD36 is associated with the insulin signaling pathway involving nPKCs.     

Restoring <Emphasis Type="Italic">Daphnia lacustris</Emphasis> G.O. Sars, 1862 (Crustacea,Anomopoda): a cryptic species in the <Emphasis Type="Italic">Daphnia longispina</Emphasis> group

While molecular markers have revealed several distinct species within the Daphnia longispina group, there is a need to reconcile these species with traditional nomenclature. Here we show that one such species, called D. longispina in recent literature based on molecular markers, can reliably be associated with the described taxon Daphnia lacustris G.O. Sars, 1862. Both mitochondrial and nuclear molecular markers readily distinguish this species from others in the D. longispina group. D. lacustris is absent in the region from which D. longispina was first described (Denmark), and the designation D. longispina must be reserved for another widespread species represented by Danish lineages. While the diagnosis of D. lacustris (and other species of the D. longispina group) by molecular markers is unequivocal, distinguishing it morphologically from other species is still problematic. The presently known distribution range of D. lacustris includes most of Norway, northern Finland and a single lake in the Polish Tatra Mountains. Its typical habitat is oligotrophic lakes without intense fish predation. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

Agricultural intensification and farmland birds: new insights from a central European country

The relationship between agricultural intensification and a decline in farmland bird populations is well documented in Europe, but the results are mostly based on data from the western part of the continent. In the former socialist eastern and central European countries, political changes around 1990 resulted in a steep decline in the intensity of agriculture. Therefore, one would expect populations of farmland birds to have recovered under these conditions of lower agricultural intensity. We explored population trends of 19 farmland bird species in the Czech Republic between 1982 and 2003 using data from a large‐scale monitoring scheme, and, additionally, we looked for relationships between such population changes and a number of variables describing the temporal development of Czech agriculture. Most farmland species declined during the focal period, and this decline was steepest in farmland specialists (Northern Lapwing Vanellus vanellus, Skylark Alauda arvensis, Linnet Carduelis cannabina and Yellowhammer Emberiza citrinella). Although the intensity of agriculture was lower after than before 1990, the overall decline continued in most farmland bird species, albeit at a slower rate. The correlations between agricultural intensity and farmland bird decline showed opposite patterns to that found in other European studies, because bird populations were highest in years with the most intensive agriculture. We speculate that this pattern could have resulted from the impact of different driving forces causing farmland bird decline in different periods. The high intensity of agriculture could have caused the decline of the originally abundant populations before 1990. After 1990, the decreasing area of arable land could be the most important factor resulting in the continued decline of farmland bird populations. Our results demonstrate that the drivers of farmland bird population changes could differ across Europe, and thus investigations into the effect of farmland management in different parts of the continent are urgently required.