Crystal Structures of α-Crystallin Domain Dimers of αB-Crystallin and Hsp20

Small heat shock proteins (sHsps) are a family of large and dynamic oligomers highly expressed in long-lived cells of muscle, lens and brain. Several family members are upregulated during stress, and some are strongly cytoprotective. Their polydispersity has hindered high-resolution structure analyses, particularly for vertebrate sHsps. Here, crystal structures of excised α-crystallin domain from rat Hsp20 and that from human αB-crystallin show that they form homodimers with a shared groove at the interface by extending a β sheet. However, the two dimers differ in the register of their interfaces. The dimers have empty pockets that in large assemblies will likely be filled by hydrophobic sequence motifs from partner chains. In the Hsp20 dimer, the shared groove is partially filled by peptide in polyproline II conformation. Structural homology with other sHsp crystal structures indicates that in full-length chains the groove is likely filled by an N-terminal extension. Inside the groove is a symmetry-related functionally important arginine that is mutated, or its equivalent, in family members in a range of neuromuscular diseases and cataract. Analyses of residues within the groove of the αB-crystallin interface show that it has a high density of positive charges. The disease mutant R120G α-crystallin domain dimer was found to be more stable at acidic pH, suggesting that the mutation affects the normal dynamics of sHsp assembly. The structures provide a starting point for modelling higher assembly by defining the spatial locations of grooves and pockets in a basic dimeric assembly unit. The structures provide a high-resolution view of a candidate functional state of an sHsp that could bind non-native client proteins or specific components from cytoprotective pathways. The empty pockets and groove provide a starting model for designing drugs to inhibit those sHsps that have a negative effect on cancer treatment.     

Cold-Inhibited Phloem Translocation in Sugar Beet: II. CHARACTERIZATION AND LOCALIZATION OF THE SLOW-COOLING RESPONSE

Experiments were undertaken on a simplified sugar beet systemto characterize the phloem translocation response to slow coolingtreatments that were applied to the source leaf petiole. Inthese experiments the temperature was decreased by 4°C every16 min, such that the tissue temperature was lowered from 25°Cto 1°C over a period of 80 min. Our results indicated thatan initial slow cooling treatment, on a given test plant, causedno change in the rate of translocation. However, all subsequentslow cooling regimes that were applied to the same petiole positionelicited a characteristic step-type inhibition. This inhibitionaveraged about 10% of the original translocation rate in allcases with no recovery being observed. The data suggest thatthe initial cooling treatment induced an alteration in the petioletissue which facilitated the inhibition phenomenon during subsequentslow coolings. This alteration was shown to be localized withinthe upstream region of the chilled petiole segment, followingan initial slow cooling, or throughout the chilled petiole segmentafter an initial quick cooling from 25°C to 1°C. Resultsalso show that the alteration is a long-lived phenomenon thathas no detectable influence on the quick-cooling induced transientinhibition of translocation. Key words: Phloem, Translocation, Cooling response, Petiole     

Survival strategies in polar terrestrial arthropods

Three components of the survival strategy of a terrestrial Antarctic mite, Alaskozetes antarcticus (Acari: Cryptostigmata) are considered: overwintering survival, energetics and life history. Supercooling is an important feature of its cold tolerance, whilst elevation of standard metabolism allows activity at low temperatures, both of which contribute tcTa long development and maximum survival of individuals in the population. These are facets of the overall survival strategy evolved by such a species in response to the Antarctic terrestrial environment, but which may be widespread in polar invertebrates.     

The internal phylogeny of ants (Hymenoptera: Formicidae)

Abstract. The higher phylogeny of the Formicidae was analysed using 68 characters and 19 taxa: the 14 currently recognized ant subfamilies plus 5 potentially critical infrasubfamilial taxa. The results justified the recognition of 3 additional subfamilies: Aenictogitoninae Ashmead (new status), Apomyrminae Dlussky & Fedoseeva (new status), and Leptanilloidinae Bolton (new subfamily). A second analysis on these better delimited 17 subfamilies resulted in 24 equally most parsimonious trees. All trees showed a basal division of extant Formicidae into two groups, the first containing (Myrmicinae, Pseudomyrmecinae, Nothomyrmeciinae, Myrmeciinae, Formicinae, Dolichoderinae, Aneuretinae) and the second the remaining subfamilies. Clades appearing within these groups included the Cerapachyinae plus 'army ants', the Nothomyrmeciinae plus Myrmeciinae, the 'formicoid' subfamilies (Aneuretinae + Dolichoderinae + Formicinae), and the Old World army ants (Aenictinae + Aenictogitoninae + Doryline), but relationships within the last two groups were not resolved, and the relative positions of the Apomyrminae, Leptanillinae and Ponerinae remained ambiguous. Moreover, a bootstrap analysis produced a consensus tree in which all branches were represented in proportions much lower than 95%. A reconstruction of the ground plan of the Formicidae indicated that the most specialized of all recent ants are the members of the subfamily Dorylinae and the least specialized ones are the monotypic Apomyrminae.     

Two New Subspecies of Herpetomonas muscarum (Leidy, 1856) Kent, 1880

SYNOPSIS. Herpetomonas muscarum muscarum n. subsp. was isolated from Musca domestica L. In culture at 20 C it assumed the opisthomastigote (up to 15%), double-flagellate and flagellate promastigote forms. At 30 C or with 4% urea added to cultures at 20 C, the proportion of opisthomastigotes was greater (up to 40%). In experimentally infected flies only transient infections, which included both opisthomastigotes and promastigotes, occurred. The promastigotes were 15–30 μ long and the kinetoplast was small and subspherical or transversely elongate. H. muscarum ingenoplastis n. subsp. was isolated from Phormia regina (Meigen). In culture at 20 C almost all individuals were double-flagellate promastigotes 20–40 μ long and less than 1% were opisthomastigotes. At 30 C or with added urea there was no increase in the proportion of opisthomastigotes and the cultures were not vigorous. In experimentally infected flies opisthomastigotes were 5–39% of the population depending on the part of the gut sampled. In all stages the kinetoplast was large (1.5–2.5 μ long) and tear-drop-shaped with the point directed posteriorly.
In artificially mixed cultures of H. m. muscarum and H. m. ingenoplastis the former predominated after a short time and eventually survived alone. A mixed culture that was about 98% H. m. muscarum was fed to Phormia regina and produced heavy pure infections of H. m. ingenoplastis , which lasted for 22 days with no indication of decline. No evidence of cyst formation was found in either subspecies.     

Stomatal Response to Humidity and Lanthanum

Lanthanum fed to the base of excised leaves of Sesamum indicum L. and Helianthus annuus L. was used as a tracer to investigate by electron microscopy the path of water in the apoplast of leaves. The generally random distribution of lanthanum in cell walls provided no support for the hypothesis that cuticular transpiration may be greater for guard cells than for adjacent epidermal cells. Occasionally, accumulations of lanthanum were observed in anticlinal walls of epidermal cells and at the outer surface of the plasma membrane but lanthanum was not observed in the symplast. The influx of ⁸⁶Rb to excised roots of sesame and sunflower was inhibited during incubation with 0.5 mM lanthanum or calcium for 15 or for 180 min. Stomata of sunflower partially closed when 2.5 mM lanthanum was supplied to the base of excised shoots in a potometer, whereas this treatment had little effect on stomatal conductance of sesame shoots maintained in a constant environment. Supplying 2.5 mM lanthanum to the base of sesame shoots strongly inhibited stomatal opening response to increase in ambient humidity but had little effect on stomatal opening response to light. It was concluded that stomatal opening response to increased humidity may be dependent upon some process, such as ion influx, that is inhibited by lanthanum, and that opening response to humidity may differ in mechanism from stomatal opening response to increased irradiance.