Genetic structure of the stonefly, Yoraperla brevis, populations: the extent of gene flow among adjacent montane streams

1. Gene flow in populations of stream insects is expected to depend on the distance between and the connectedness of sites in stream networks, and on dispersal ability (i.e. larval drift and adult flight).
2. Yoraperla brevis (Banks) is an abundant and characteristic stonefly of smaller streams in the northern Rocky Mountains. The present authors analysed genetic structure at 27 sites in sevenz streams flowing into the Bitterroot River in western Montana, USA. Cellulose acetate electrophoresis identified five variable loci with 16 alleles.
3. Genotype frequencies conformed to Hardy–Weinberg expectations. Within-stream differentiation was low and among-stream variation ( F _st) was an order of magnitude higher.
4. UPGMA grouped sites within streams and also grouped adjacent streams. The tree produced by the Neighbour Joining Method was similar although not quite so clear cut.
5. This orderly pattern (i.e. Hardy–Weinberg proportions, homogeneity within streams and geographical structure) contrasts strongly with patterns observed in invertebrates from subtropical streams in Australia. Yoraperla brevis maintains large populations in predictable environments, has a long life-cycle with a likelihood of cohort mixing, emerges synchronously in large breeding populations and occupies streams separated by areas of high relief; the Australian situation is the opposite in most respects.
6. Further analysis of a range of species is required to determine whether the different genetic structure in Y. brevis compared to the Australian species occurs more generally in North American stream insects.     

Human p38α mitogen-activated protein kinase in the Asp168-Phe169-Gly170-in (DFG-in) state can bind allosteric inhibitor Doramapimod

Doramapimod (BIRB-796) is widely recognized as one of the most potent and selective type II inhibitors of human p38α mitogen-activated protein kinase (MAPK); however, the understanding of its binding mechanism remains incomplete. Previous studies indicated high affinity of the ligand to a so-called allosteric pocket revealed only in the ‘out’ state of the DFG motif (i.e. Asp168-Phe169-Gly170) when Phe169 becomes fully exposed to the solvent. The possibility of alternative binding in the DFG-in state was hypothesized, but the molecular mechanism was not known. Methods of bioinformatics, docking and long-time scale classical and accelerated molecular dynamics have been applied to study the interaction of Doramapimod with the human p38α MAPK. It was shown that Doramapimod can bind to the protein even when the Phe169 is fully buried inside the allosteric pocket and the kinase activation loop is in the DFG-in state. Orientation of the inhibitor in such a complex is significantly different from that in the known crystallographic complex formed by the kinase in the DFG-out state; however, the Doramapimod’s binding is followed by the ligand-induced conformational changes, which finally improve accommodation of the inhibitor. Molecular modelling has confirmed that Doramapimod combines the features of type I and II inhibitors of p38α MAPK, i.e. can directly and indirectly compete with the ATP binding. It can be concluded that optimization of the initial binding in the DFG-in state and the final accommodation in the DFG-out state should be both considered at designing novel efficient type II inhibitors of MAPK and homologous proteins.

Communicated by Ramaswamy H. Sarma     

Crystal structure of a NO-forming nitrite reductase mutant: an analog of a transition state in enzymatic reaction

I257E was obtained by site directed mutagenesis of nitrite reductase from Achromobacter cycloclastes. The mutant has no enzyme activity. Its crystal structure determined at 1.65A resolution shows that the side-chain carboxyl group of the mutated residue, Glu257, coordinates with the type 2 copper in the mutant and blocks the contact between the type 2 copper and its solvent channel, indicating that the accessibility of the type 2 copper is essential for maintaining the activity of nitrite reductase. The carboxylate is an analog of the substrate, nitrite, but the distances between the type 2 copper and the two oxygen atoms of the side-chain carboxyl group are reversed in comparison to the binding of nitrite to the native enzyme. In the mutant, both the type 2 copper and the N epsilon atom on the imidazole ring of its coordinated residue His135 move in the substrate binding direction relative to the native enzyme. In addition, an EPR study showed that the type 2 copper in the mutant is in a reduced state. We propose that mutant I257E is in a state corresponding to a transition state in the enzymatic reaction.     

The relationship between sequence and interaction divergence in proteins

There is currently a gap in knowledge between complexes of known three-dimensional structure and those known from other experimental methods such as affinity purifications or the two-hybrid system. This gap can sometimes be bridged by methods that extrapolate interaction information from one complex structure to homologues of the interacting proteins. To do this, it is important to know if and when proteins of the same type (e.g. family, superfamily or fold) interact in the same way. Here, we study interactions of known structure to address this question. We found all instances within the structural classification of proteins database of the same domain pairs interacting in different complexes, and then compared them with a simple measure (interaction RMSD). When plotted against sequence similarity we find that close homologues (30-40% or higher sequence identity) almost invariably interact the same way. Conversely, similarity only in fold (i.e. without additional evidence for a common ancestor) is only rarely associated with a similarity in interaction. The results suggest that there is a twilight zone of sequence similarity where it is not possible to say whether or not domains will interact similarly. We also discuss the rare instances of fold similarities interacting the same way, and those where obviously homologous proteins interact differently.     

Patch occupancy of two hemipterans sharing a common host plant

Aim Two hemipteran species were chosen as a study system for the comparative analysis of patch occupancy and spatial population structure of insects sharing a common host plant. This study tested whether (1) the incidence in the host plant patches differed between the two species, and (2) the two species exhibited a different spatial population structure, i.e. were they affected differentially by isolation and area of the host plant patches. Location The porphyry landscape north of Halle (Saale) in Germany comprising 506 patches of the host plant Brachypodium pinnatum. Methods The host plant patches were surveyed for the two hemipterans. To assess the influence of patch quality on species occurrence the patches were characterized by mean cover abundance of B. pinnatum, type of subsoil, slope, exposure, and shading. The spatial configuration of the patches was considered by patch area and isolation. The influence of the habitat factors and the spatial configuration on the occupancy of the two species was analysed by logistic regression. Results Adarrus multinotatus was found in 441 patches, while Neophilaenus albipennis was found in only 90 patches. While A. multinotatus showed virtually no relationship to the habitat factors, the occupancy of N. albipennis was influenced by subsoil type, cover abundance, and shading. The effects of area and isolation on occupancy of the patches also differed between the two species. The occupancy of N. albipennis was determined largely by area and isolation, whereas in A. multinotatus no considerable effect of spatial configuration was found. Main conclusions The study revealed a marked difference between the two hemipteran species in respect of spatial population structure. Adarrus multinotatus built up a ‘patchy population’, whereas N. albipennis showed a ‘metapopulation’ structure within the same set of patches in the same landscape. Spatial population structure was found to be not only a function of spatial configuration of habitat patches, but population structure differed between the habitat generalist A. multinotatus and the habitat specialist N. albipennis.     

Synthesis of magnesium titanate nanocrystallites from a cheap and water-soluble single source precursor

Pure nanocrystallite magnesium titanate (MgTiO₃) was conveniently synthesized by thermal decomposition of a cheap and water-soluble heterobimetallic single source precursor [Mg(H₂O)₅]₂[Ti₂(O₂)₂O(NC₆H₆O₆)₂]·7H₂O at low temperature. This single source precursor was obtained in high yield and in a crystalline form from the quaternary system of MgO-Ti(OC₄H₉)₄-H₂O₂-H₃nta (H₃nta = nitrilotriacetic acid) at pH 4.0. It was characterized by elemental analysis, IR spectrum, NMR, thermal gravimetric analysis and X-ray single-crystal diffraction. The morphology, microstructure, and crystallinity of the resulting MgTiO₃ materials have been characterized by transmission electron microscopy and X-ray diffraction. The TEM image of the resulting MgTiO₃ powders only consists of the nano-scale crystallites with the crystalline size of 30-100 nm.